Rabdosia rubescens / dong ling cao

The cytostatic and cytotoxic effects of oridonin (Rubescenin), a diterpenoid from Rabdosia rubescens, on tumor cells of different lineage.

Chen S, Gao J, Halicka HD, Huang X, Traganos F, Darzynkiewicz Z. Int J Oncol. 2005 Mar;26(3):579-88.

Rabdosia rubescens is a herbal medicine used to treat esophageal cancer in China. In this study, the sesquiterpene oridonin, an isoprenoid, was isolated from Rabdosia rubescens. Mass spectroscopy and carbon 13 NMR spectroscopy were used to identify the structure of the purified compound. It was then evaluated for biological activity against human cell lines derived from prostate (DU-145, LNCaP), breast (MCF-7), and ovarian (A2780 and PTX10) cancers. Oridonin exhibited anti-proliferative activity toward all cancer cell lines tested, with an IC50 estimated by the MTT cell viability assay ranging from 5.8+/-2.3 to 11.72+/-4.8 microM. Flow cytometric analysis demonstrated that oridonin induced a G1 phase arrest in androgen receptor-positive LNCaP cells containing wt p53, while it blocked the cell cycle at G2 and M phases in androgen receptor-negative DU-145 cells with mutated p53; the arrest in M was verified by examination of cell morphology and by the increased frequency of cells with Ser-10 phosphorylated histone H3. The increased incidence of apoptosis, identified by characteristic changes in cell morphology, was seen in tumor lines treated with oridonin. Notably, at concentrations that induced apoptosis among tumor cells, oridonin failed to induce apoptosis in cultures of normal human fibroblasts. Western blot analysis was used to determine the protein expression of cancer suppressor genes, p53 (wt) and Bax, and the proto-oncogene, Bcl-2 in LNCaP cells following treatment with oridonin. Oridonin up-regulated p53 and Bax and down-regulated Bcl-2 expression in a dose-dependent manner. To further explore the possible interaction between oridonin and DNA, its absorption spectrum was measured in the presence and absence of double stranded (ds) DNA. Spectral shifts and an increase in absorption band intensity were observed indicating interaction of oridonin with DNA bases. The nature of the binding is not clear at present though no evidence of histone H2AX phosphorylation on Ser-139 was apparent in DU-145 cells treated with oridonin that would indicate the induction of ds DNA breaks. In conclusion, oridonin inhibits cancer cell growth in a cell cycle specific manner and shifts the balance between pro- and anti-apoptotic proteins in favor of apoptosis. The present data suggest that further studies are warranted to assess the potential of oridonin in cancer prevention and/or treatment.

Ponicidin and oridonin are responsible for the antiangiogenic activity of Rabdosia rubescens, a constituent of the herbal supplement PC SPES.

Meade-Tollin LC, Wijeratne EM, Cooper D, Guild M, Jon E, Fritz A, Zhou GX, Whitesell L, Liang JY, Gunatilaka AA. J Nat Prod. 2004 Jan;67(1):2-4.

Antiangiogenic activity has been identified in an aqueous EtOH extract of Rabdosia rubescens, a component of the dietary supplement PC SPES. Bioassay-guided fractionation using a novel in vitro human endothelial cell-based assay for angiogenesis afforded the diterpenoids ponicidin (1) and oridonin (2), with significant antiangiogenic activity at subcytotoxic concentrations, suggesting that these constituents may strongly contribute to the demonstrated clinical efficacy of PC SPES as a treatment for advanced prostate cancer.

Oridonin induces growth inhibition and apoptosis of a variety of human cancer cells.

Ikezoe T, Chen SS, Tong XJ, Heber D, Taguchi H, Koeffler HP. Int J Oncol. 2003 Oct;23(4):1187-93.

PC-SPES is an eight herbal mixture that was shown to have activity against prostate cancer. Recently, we purified oridonin from Rabdosia rubescens, one component of PC-SPES, by high performance liquid chromatography (HPLC). The ability of oridonin to inhibit the proliferation of cancer cells was examined by MTT assay. Oridonin effectively inhibited the proliferation of a wide variety of cancer cells including those from prostate (LNCaP, DU145, PC3), breast (MCF-7, MDA-MB231), non-small cell lung (NSCL) (NCI-H520, NCI-H460, NCI-H1299) cancers, acute promyelocytic leukemia (NB4), and glioblastoma multiforme (U118, U138) with ED50s ranging from 1.8 to 7.5 micro g/ml. TUNEL assay and cell cycle analysis showed that oridonin induced apoptosis and G0/G1 cell cycle arrest in LNCaP prostate cancer cells. In addition, expression of p21waf1 was induced in LNCaP and NCI-H520 cells in a p53-dependent manner. Interestingly, when p53 was suppressed by over-expression of E6 from human papilloma virus type 16 (HPV-16), these cells lost their sensitivity to oridonin-induced growth inhibition and apoptosis. Taken together, oridonin inhibited the proliferation of cancer cells via apoptosis and cell cycle arrest with p53 playing a central role in several cancer types, which express the wild-type p53 gene. Oridonin may be a novel, adjunctive therapy for a large variety of malignancies and probably represents one of the major, active components of PC-SPES.

Wedelia chinensis/Peng qi ju

Compounds from Wedelia chinensis synergistically suppress androgen activity and growth in prostate cancer cells.

Lin FM, Chen LR, Lin EH, et al. Carcinogenesis. 2007 Dec;28(12):2521-9.

Chronic inflammation can augment tumor development in various types of cancers, including prostate cancer (PCa). Reduction of inflammation is therefore an important anticancer therapeutic opportunity. Here, we report four anti-proliferative phytocompounds in Wedelia chinensis, an oriental herbal medicine, identified through their ability to modulate the androgen receptor (AR) activation of transcription from prostate-specific antigen promoter in PCa cells. The 50% inhibition concentration values of indole-3-carboxylaldehyde, wedelolactone, luteolin and apigenin, were 34.9, 0.2, 2.4 and 9.8 muM, respectively. A formula that combined the phytocompounds in the same proportions as in the herbal extract decreased the dosage of each compound required to achieve maximal AR inhibition.

In correlation with the AR suppression effect, these active compounds specifically inhibited the growth of AR-dependent PCa cells and as a combination formula they also synergistically suppressed growth in AR-dependent PCa cells. Our study has identified synergistic effects of active compounds in W. chinensis and demonstrated their potential in PCa prevention and therapy. The paradigm of multiple activities and synergism is a useful framework to investigate the therapeutic effects of whole extracts from assorted medicinal plant species.

Herbal extract of Wedelia chinensis attenuates androgen receptor activity and orthotopic growth of prostate cancer in nude mice.

Tsai CH, Lin FM, Yang YC, Lee MT, Cha TL, Wu GJ, Hsieh SC, Hsiao PW. Clin Cancer Res. 2009 Sep 1;15(17):5435-44. Epub 2009 Aug 18.

PURPOSE: Wedelia chinensis is a common ingredient of anti-inflammatory herbal medicines in Taiwan and southern China. Inflammation is involved in promoting tumor growth, invasion, and metastasis. This study aims to test the biological effects in vivo of W. chinensis extract on prostate cancer. EXPERIMENTAL DESIGN: The in vivo efficacy and mechanisms of action of oral administration of a standardized extract of W. chinensis were analyzed in animals bearing a subcutaneous or orthotopic prostate cancer xenograft. RESULTS: Exposure of prostate cancer cells to W. chinensis extract induced apoptosis selectively in androgen receptor (AR)-positive prostate cancer cells and shifted the proportion in each phase of cell cycle toward G(2)-M phase in AR-negative prostate cancer cells. Oral herbal extract (4 or 40 mg/kg/d for 24-28 days) attenuated the growth of prostate tumors in nude mice implanted at both subcutaneous (31% and 44%, respectively) and orthotopic (49% and 49%, respectively) sites. The tumor suppression effects were associated with increased apoptosis and lower proliferation in tumor cells as well as reduced tumor angiogenesis. The antitumor effect of W. chinensis extract was correlated with accumulation of the principal active compounds wedelolactone, luteolin, and apigenin in vivo. CONCLUSION: Anticancer action of W. chinensis extract was due to three active compounds that inhibit the AR signaling pathway. Oral administration of W. chinensis extract impeded prostate cancer tumorigenesis. Future studies of W. chinensis for chemoprevention or complementary medicine against prostate cancer in humans are thus warranted.

Compounds from Wedelia chinensis synergistically suppress androgen activity and growth in prostate cancer cells.

Lin FM, Chen LR, Lin EH, Ke FC, Chen HY, Tsai MJ, Hsiao PW. Carcinogenesis. 2007 Dec;28(12):2521-9. Epub 2007 Oct 17.

Chronic inflammation can augment tumor development in various types of cancers, including prostate cancer (PCa). Reduction of inflammation is therefore an important anticancer therapeutic opportunity. Here, we report four anti-proliferative phytocompounds in Wedelia chinensis, an oriental herbal medicine, identified through their ability to modulate the androgen receptor (AR) activation of transcription from prostate-specific antigen promoter in PCa cells. The 50% inhibition concentration values of indole-3-carboxylaldehyde, wedelolactone, luteolin and apigenin, were 34.9, 0.2, 2.4 and 9.8 muM, respectively. A formula that combined the phytocompounds in the same proportions as in the herbal extract decreased the dosage of each compound required to achieve maximal AR inhibition. In correlation with the AR suppression effect, these active compounds specifically inhibited the growth of AR-dependent PCa cells and as a combination formula they also synergistically suppressed growth in AR-dependent PCa cells. Our study has identified synergistic effects of active compounds in W. chinensis and demonstrated their potential in PCa prevention and therapy. The paradigm of multiple activities and synergism is a useful framework to investigate the therapeutic effects of whole extracts from assorted medicinal plant species.

Scutellariae / Huang qin

Molecular mechanism of anti-prostate cancer activity of Scutellaria baicalensis extract.

Ye F, Jiang S, Volshonok H, Wu J, Zhang DY. Nutr Cancer. 2007;57(1):100-10.

Scutellaria baicalensis is a widely used Chinese herbal medicine historically used in anti-inflammatory and anticancer therapy. The goals of the study were to 1) determine its in vitro and in vivo anti-prostate cancer activity, 2) investigate its molecular mechanism directed at cell proliferation control including cyclooxygenase-2(COX-2) prostaglandin E2 (PGE2) and cyclins/cdks pathways, and 3) compare it with those of PC-SPES (PC stands for prostate cancer and spes is Latin for hope), a former herbal mixture for prostate cancer treatment of which S. baicalensis is a major constituent. Two human prostate cancer cell lines (LNCaP, androgen dependent, and PC-3, androgen independent) were assessed for growth inhibition. S. baicalensis exerted dose- and time-dependent increased growth inhibition in both cell lines. However, the PC-3 cells IC50 (50% growth inhibition concentration) were slightly more sensitive than LNCaP cells (IC50=0.15 mg/ml), although the former is androgen independent. S. baicalensis was more effective in inhibition of cell growth compared with PC-SPES (IC50=0.38 mg/ml for PC-3 cells). Significant reduction of PGE2 synthesis in both cells after treatment with S. baicalensis resulted from direct inhibition of COX-2 activity rather than COX-2 protein suppression. S. baicalensis also inhibited prostate-specific antigen production in LNCaP cells. Finally, S. baicalensis suppressed expression of cyclin D1 in LNCaP cells, resulting in a G1 phase arrest, while inhibiting cdk1 expression and kinase activity in PC-3 cells, ultimately leading to a G2/M cell cycle arrest. Animal studies showed a 50% reduction in tumor volume after a 7-wk treatment period. This study demonstrated that S. baicalensis may be a novel anticancer agent for the treatment of prostate cancer.

Characterization of chemical constituents in Scutellaria baicalensis with anti-androgenic and growth-inhibitory activities toward prostate carcinoma.

Bonham M, Posakony J, Coleman I, Montgomery B, Simon J, Nelson PS. Clin Cancer Res. 2005 May 15;11(10):3905-14.

PURPOSE: Botanical preparations are widely used by patients with prostate cancer. Scutellaria baicalensis, a botanical with a long history of medicinal use in China, was a constituent of the herbal mixture PC-SPES, a product that inhibited prostate cancer growth in both laboratory and clinical studies. Due to the difficulties encountered when evaluating the efficacy of complex natural products, we sought to identify active chemical constituents within Scutellaria and determine their mechanisms of action. EXPERIMENTAL DESIGN AND RESULTS: We used high-performance liquid chromatography to fractionate S. baicalensis and identified four compounds capable of inhibiting prostate cancer cell proliferation; baicalein, wogonin, neobaicalein, and skullcap flavone. Comparisons of the cellular effects induced by the entire extract versus the four-compound combination produced comparable cell cycle changes, levels of growth inhibition, and global gene expression profiles (r(2) = 0.79). Individual compounds exhibited anti-androgenic activities with reduced expression of the androgen receptor and androgen-regulated genes. In vivo, baicalein (20 mg/kg/d p.o.) reduced the growth of prostate cancer xenografts in nude mice by 55% at 2 weeks compared with placebo and delayed the average time for tumors to achieve a volume of approximately 1,000 mm(3) from 16 to 47 days (P < 0.001). CONCLUSIONS: Most of the anticancer activities of S. baicalensis can be recapitulated with four purified constituents that function in part through inhibition of the androgen receptor-signaling pathway. We conclude that clinical studies evaluating the efficacy of these agents in the context of chemoprevention or the treatment of prostate cancer are warranted.

Study of baicalin in inducing prostate cancer cell line DU145 apoptosis in vitro.

Gu ZQ, Sun YH, Xu CL, Liu Y. Zhongguo Zhong Yao Za Zhi. 2005 Jan;30(1):63-6.

OBJECTIVE: The purpose of the present study was to investigate the in vitro effects of baicalin on induction of apoptosis in human prostate cancer cell line DU145. METHOD: Human prostate cancer cell line DU145 was treated with different concentration of baicalin in vitro. The apoptosis rate was determined by FACS analysis, cell cycle distribution was detected by flow cytometry, morphological changes and protein analysis were determined by means of electron microscope technique and immunohistochemical technique respectively. RESULT: 50micromol x L(-1) and 125 micromol x L(-1) of baicalin dose-dependently induced apoptosis and inhibited the proliferation of prostate cancer cell DU145 in a dose and time-dependent manner. DNA flow cytometric analysis indicated that baicalin induced a arrest in G1 phase, showing a typical apoptosis peak. Electron microscopy detected a characteristic appearance of the apoptotic cells morphology. Immunohistochemical analysis revealed that induction of apoptosis by ways of inhibition of the bcl-2, loss of the Bax, and up regulation of Fas. CONCLUSION: The results indicate that baicalin may induce apoptosis and inhibit proliferation of prostate cancer cells, and has direct anti-tumor effects on human prostate cancer cells.

The effect of baicalin on proliferation, apoptosis and differentiation of human hepatocellular line BEL-7402 in vitro

Guo Yu, Guo Xia, Yao Jinfeng, Wang Na. Zhong Yao Yao Li Yu Lin Chuang. 2008; 24(3): 21-23.

Objective: To study the effect of baicalin on proliferation, apoptosis and differentiation of human hepatocellular line BEL-7402. Methods: Human hepatocellular line BEL-7402 were cultured in vitro. Cell morphology and ultrastructure was examined by a light and electronic microscope. The specific activities of alkaline phosphatase (ALP) and heat-resistant ALP were assayed by ALP kit based on Bessey method. The specific activity of gamma-glutamyl transferase (g-GT) was measured with g-GT kit. The secretory amount of AFP, albumin and cAMP was detected with radioimmunoassay kit. The effects of baicalin on apoptosis were observed by FCM. Results: Baicalin inhibited proliferation and promoted apoptosis of hepatocarcinoma cell BEL-7402 in a time-dose dependent manner. Baicalin induced the mature and normality of morphology and ultrastructure. After treated by baicalin g-GT activity descended significantly, ALP activity were remarkably enhanced, secretory amount of AFP dropped, albumin secretory amount ascended. Conclusion: Baicalin could inhibit proliferation, induce the BEL-7402 cell differentiation tending to normal. Subject words hepatocellular carcinoma; baicalin; proliferation; apoptosis; differentiation.

Effect and mechanism of baicalin on proliferation, invasion and metastasis of human hepatocellular line bel-7402 in vitro.

Guo Yu, Guo Xia, Wu Jingxuea, Yao Jinfeng Cui Donglai. Zhong Guo Zu Zhi Hua Xue Yu Xi Bao Hua Xue Za Zhi. 2008; 17(4): 329-334.

Objective To study the effect and mechanism of baicalin on proliferation, invasion and metastasis of human hepatocellular line BEL-7402. Methods Human hepatocellular line BEL-7402 was cultured in vitro. MTT and the soft agar colony experiment were used to evaluate cell growth. Invasion assay was conducted in the Boyden chamber, while cell migration and morphology was also observed. The expression of MMP2 and TIMP2 proteins of human hepatocellular line EEL-7402 was determined by flow cytometry. The expression of VEGF was determined by immunohistochemical techniques. Results Baicalin inhibited proliferation, invasion and migration of hepatocellular EEL-7402. Morphologically, the cells treated with baicalin were round with relatively fewer pseudopods. Meanwhile, MMP2 expression was decreased and TIMP2 expression was increased significantly, so MMP2/TIMP2 ratio was significantly decreased. VEGF expression was also decreased obviously. Conclusion Baicalin can inhibit the proliferation, invasion and metastasis of human hepatocellular line BEL-7402, which may be the results of decease in MMP2/TIMP2 ratio and VEGF expression.

Antitumor effects of Baicalin and its effect on telomerase in tumor cells

LI Hongjie, XIE Wenli, ZHU Jiang. Jiang Su Yi Yao. 2008; 34(9): 931-933.

Objective: To study the inhibitory effect of Baicalin on tumor cells and its influence on telomerase activity in Eca-109. Methods: The inhibitory effect of bacailin on the proliferation of three kinds of human tumor cells was assayed by MTT method. Transplant tumor models of S180 and H22 were used. After transplantation the inhibitory rates were calculated. Results: Baeailin 20-100 mg/ml inhibited the proliferation of three kinds of tumor cells in vitro in a dose-dependent manner (P<0.01). The inhibitory rates of bacailin (5, 10, 20 mg/kg IP) in vivo were 29.9%-71.5% and 35.1 %-80.6% in tumor-bearing mice with S180 and H22, respectively. Conclusion: Bacailin could inhibit the proliferation of three kinds of tumor cells in vitro and presents antitumor effect on the tumor-bearing mice in vivo. Bacailin can effectively inhibit telomere activity of turner cells.

Inhibition of Baicalin on Proliferation of Hepatoma HepG -2 Cell Line

ZHANG Xue-wu, CUI Chang-xu, CHEN Li-yan, QUAN Ji-shu. Shi Zhen Guo Yi Guo Yao. 2008; 19(9): 2154-2155.

Objective: To investigate the inhibition of Baicalin on proliferation hepatoma HepG-2 cells. Methods: MTr assay was used to examine the effect of baicalin on the proliferation of HepG-2 cell and flow cytometry was used for the cell cycle distribution. Results: Baicalin inhibited the proliferation of HepG-2 cell and the effects were in a time-and-concentration dependent manner. Baicalin could also induce apoptosis, the S phase was arrested and the ratio of apoptosis there was significant difference than that of control group, respectively. Conclusion: Baicalin inhibits proliferation of HepG-2 cells via induction of apoptosis and cell cycle arrest.

Inhibitory effect of baicalein on human cervix cancer HeLa cells

TONG Xu-hui, DONG Shu-ying, TAO Liang. Beng Bu Yi Xue Yuan Xue Bao. 2009; 34(6): 468-470.

Objective: To investigate the role of baicalein in inhibiting the HeLa cells of human cervix cancer and the possible mechanism. Methods: Sulforhodamine B (SRB) assay was used to determine the inhibitory effect of baicalein on HeLa cell growth; cellular apoptosis was detected by Hoechst 33258 fluorescent staining; caspase-3 activity was assayed by colorimetric method. Results: After treated with different concentration of baicalein for 24, 36, 48hour, HeLa cell growth were significantly inhibited (P<0.05 to P<0.01); the apoptosis rate of HeLa cells(P<0.01) and the activity of caspase-3 (P<0.05 to P<0.01) were increased when HeLa cells were treated with baicalein for 24 hours. Conclusions: Baicalein can inhibit the growth of HeLa cells, and the inhibitory effect probably results from the increased cell apoptosis and caspase-3 activity.

Indirubin

Indirubin derivatives inhibit Stat3 signaling and induce apoptosis in human cancer cells.

Nam S, Buettner R, Turkson J, Kim D, Cheng JQ, Muehlbeyer S, Hippe F, Vatter S, Merz KH, Eisenbrand G, Jove R. Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):5998-6003. Epub 2005 Apr 18.

Stat3 protein has an important role in oncogenesis and is a promising anticancer target. Indirubin, the active component of a traditional Chinese herbal medicine, has been shown previously to inhibit cyclin-dependent kinases, resulting in cell cycle arrest. Here, we show that the indirubin derivatives E564, E728, and E804 potently block constitutive Stat3 signaling in human breast and prostate cancer cells. In addition, E804 directly inhibits Src kinase activity (IC(50) = 0.43 microM) in an in vitro kinase assay. Levels of tyrosyl phosphorylation of c-Src are also reduced in cultured cells 30 min after E804 treatment. Tyrosyl phosphorylation of Stat3, which is known to be phosphorylated by c-Src, was decreased, and constitutive Stat3 DNA binding-activity was suppressed in cells 30 min after E804 treatment. The antiapoptotic proteins Mcl-1 and Survivin, which are encoded in target genes of Stat3, were down regulated by indirubin derivatives, followed by induction of apoptosis. These results demonstrate that E804 directly blocks the Src-Stat3 signaling pathway, suggesting that the antitumor activity of indirubin compounds is at least partially due to inhibition of this pathway.

Pseudo ginseng / san qi

San Qi is a Chinese herb usually indicated in the treatment of pain syndrome, chronic renal failure, prostatic hyperplasia, bleeding due to ulcer, ulcerative colitis and enteritis, etc.

Prostatic hyperplasia

Hua Yu San: san qi, wang buliuxing, chuanshanjia, zhechong, hu po, 50g each were grounded into powder and capsulated, 0.5g each capsule, 6 capsules each time, tid. 15 days as a course of treatment, 2~3 courses totally. 120 cases of prostatic hyperplasia were treated and 36 markedly effective, 68 effective and 16 ineffective.

Pain in liver cancer of late stage

Xin Guan Pian (san qi, niu huang, zhong jie feng, etc.), once every 6~8 hours, 3~4 tablets each time. 22 cases were treated, and 16 were completely relieved, 5 relieved moderately, 1 ineffective.

Chrysanthemi / Ju hua

Role of herbal compounds (PC-SPES) in hormone-refractory prostate cancer: two case reports.

de la Taille A, Hayek OR, Burchardt M, Burchardt T, Katz AE. J Altern Complement Med. 2000 Oct;6(5):449-51.

PURPOSE: Herbal therapies are unconventional treatments that have been used for several different diseases. PC-SPES is an herbal mixture, composed of eight different herbs (chrysanthemum, isatis, licorice, Ganoderma lucidum, Panax pseudo-ginseng, Rabdosia rubescens, saw palmetto, and scutellaria), which has been used as an alternative in the treatment of prostate cancer. We report two cases of hormone-refractory prostate cancer patients, who showed a favorable response to therapy with this herbal combination, controlling the progression of the disease. METHODS: We report two cases of biopsy proven prostate cancer patients with metastatic disease, treated with total androgen blockade, progressing to an androgen-independent status. These patients were offered traditional therapies for hormone-resistant prostate cancer, and they chose to take PC-SPES. The follow-up as well as their evolution are described. RESULTS: PC-SPES extract decreased the prostate-specific antigen (PSA) value for both patients from an initial value of 100 and 386 ng/mL to 24 and 114 ng/mL after 1 year and 4 months, respectively, remaining stable until now. No gynecomastia or hot flashes were observed in these patients and the treatment was well tolerated. CONCLUSION: PC-SPES has shown a strong estrogenic in vitro and in vivo activity as an alternative tool in the management of prostate cancer patients. These cases suggest that PC-SPES might have some potential activity against hormone-independent prostate cancers.

Constituents of Compositae plants III. Anti-tumor promoting effects and cytotoxic activity against human cancer cell lines of triterpene diols and triols from edible chrysanthemum flowers.

Ukiya M, Akihisa T, Tokuda H, Suzuki H, Mukainaka T, Ichiishi E, Yasukawa K, Kasahara Y, Nishino H. Cancer Lett. 2002 Mar 8;177(1):7-12.

Fifteen pentacyclic triterpene diols and triols, consisting of: six taraxastanes, faradiol (1), heliantriol B0 (2), heliantriol C (3), 22alpha-methoxyfaradiol (4), arnidiol (5), and faradiol alpha-epoxide (6); five oleananes, maniladiol (7), erythrodiol (8), longispinogenin (9), coflodiol (10), and heliantriol A(1) (11); two ursanes, brein (12) and uvaol (13); and two lupanes, calenduladiol (14) and heliantriol B2 (15), isolated from the non-saponifiable lipid fraction of the edible flower extract of chrysanthemum (Chrysanthemum morifolium) were evaluated for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate, in Raji cells as a primary screening test for anti-tumor-promoters. All of the compounds tested showed inhibitory effects against EBV-EA activation with potencies either comparable with or stronger than that of glycyrrhetic acid, a known natural anti-tumor-promoter. Evaluation of the cytotoxic activity of six compounds, 1-3 and 5-7, against human cancer cell lines revealed that compound 5 possesses a wide range of cytotoxicity, with GI50 values (concentration that yields 50% growth) of mostly less than 6 microM.

Recent advances in the discovery and development of flavonoids and their analogues as antitumor and anti-HIV agents.

Wang HK, Xia Y, Yang ZY, Natschke SL, Lee KH. Adv Exp Med Biol. 1998;439:191-225.

Antitumor and anti-HIV flavonoids and their analogues will be reviewed with emphasis on those discovered in our laboratory. The active antitumor compounds include the antileukemic tricin (1) and kaempferol-3-O-beta-D-glucopyranoside (2) from Wikstroemia indica, the cytotoxic hinokiflavone (3) from Rhus succedanea, and the cytotoxic isoflavone (8) from Amorpha fruticosa, two dihydroxypentamethoxyflavones (9, 10) from Polanisia dodencandra. The development of synthetic 2-phenyl-4-quinolones as potent cytotoxic antimitotic flavonoid analogues and 2-phenylthiochromen-4-ones as potent antitumor flavonoid analogues will be presented. Selected results from other laboratories and antitumor-related biological studies also will be discussed. Flavonoids have also been investigated as potential anti-HIV agents. In our laboratory, acacetin-7-O-beta-D-galactopyranoside (131) from Chrysanthemum morifolium and chrysin (102), as well as apigenin-7-O-beta-D-glucopyranoside (130), from Kummerowia striata, have been found to exhibit anti-HIV activity. In other studies, some flavonoids and related compounds have been investigated as inhibitors of HIV-1 reverse transcriptase, protease, and integrase. The isolation and structural modification of such plant-derived active principles provide a continuing source of potential antitumor and anti-HIV agents.

Ganoderma / ling zhi

The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum (AR+PCa)

Liu J, Shimizu K, Konishi F, Kumamoto S, Kondo R. Bioorg Med Chem. 2007 Jul 15;15(14):4966-72.

The anti-androgenic activity of the ethanol extract of the fruiting body of Ganoderma lucidum has been previously reported. Ganoderol B with 5alpha-reductase inhibitory activity and the ability to bind to androgen receptor (AR) can inhibit androgen-induced LNCaP cell growth and suppress regrowth of the ventral prostate induced by testosterone in rats. The down-regulation of AR signaling by ganoderol B provides an important mechanism for its anti-androgenic activity.

In view of the fact that PSA (prostatic specific antigen, a well-accepted prognostic indicator of prostate cancer) is down-regulated, an important implication of this study is that ganoderol B intervention strategy aimed at toning down the amplitude of androgen signaling could be helpful in controlling morbidity of prostate cancer. In conclusion, results suggest that ganoderol B might be useful in prostate cancer and benign prostatic hyperplasia (BPH) therapy through suppressing the function of androgen and its receptor.

The anti-androgen effect of ganoderol B isolated from the fruiting body of Ganoderma lucidum.

Liu J, Shimizu K, Konishi F, Kumamoto S, Kondo R. Bioorg Med Chem. 2007 Jul 15;15(14):4966-72. Epub 2007 Apr 25.

The anti-androgenic activity of the ethanol extract of the fruiting body of Ganoderma lucidum has been previously reported. Ganoderol B with 5alpha-reductase inhibitory activity and the ability to bind to androgen receptor (AR) can inhibit androgen-induced LNCaP cell growth and suppress regrowth of the ventral prostate induced by testosterone in rats. The down-regulation of AR signaling by ganoderol B provides an important mechanism for its anti-androgenic activity. In view of the fact that PSA (prostatic specific antigen, a well-accepted prognostic indicator of prostate cancer) is down-regulated, an important implication of this study is that ganoderol B intervention strategy aimed at toning down the amplitude of androgen signaling could be helpful in controlling morbidity of prostate cancer. In conclusion, our result suggests that ganoderol B might be useful in prostate cancer and benign prostatic hyperplasia (BPH) therapy through suppressing the function of androgen and its receptor.

Androgen receptor-dependent and -independent mechanisms mediate Ganoderma lucidum activities in LNCaP prostate cancer cells.

Zaidman BZ, Wasser SP, Nevo E, Mahajna J. Int J Oncol. 2007 Oct;31(4):959-67.

Ganoderma lucidum (Curt.:Fr.) P. Karst, a medicinal fungus, has been widely used in Asian countries for centuries to prevent or treat a variety of diseases, including cancer. However, the mechanisms responsible for the effects of G. lucidum on cancer cells remain to be elucidated. We have previously shown that ethyl acetate extract of G. lucidum inhibits LNCaP prostate cancer cell viability and proliferation. We also demonstrated that G. lucidum extract decreased androgen receptor transcriptional activity, suppressed levels of secreted prostate-specific antigen, and suppressed androgen receptor protein level. In this study we investigated the mechanisms that underlie the activities of G. lucidum crude extract and its active fraction GLF4 in LNCaP prostate cancer cells. Our data demonstrate that G. lucidum inhibits cell viability by induction of apoptosis through the extrinsic pathway that include activation of caspase-8 and caspase-3 and inhibits cell proliferation by the down-regulation of cyclin D1 expression. Furthermore, G. lucidum crude extract and fraction GLF4 interfere with androgen receptor function via competition with the natural ligand dihydrotestosterone and suppression of androgen receptor/androgen response element complex formation. These results indicate that G. lucidum extracts have profound activity against LNCaP cells that merits further investigation as a potential therapeutic agent for the treatment of prostate cancer.

Ganoderma lucidum suppresses angiogenesis through the inhibition of secretion of VEGF and TGF-beta1 from prostate cancer cells.

Stanley G, Harvey K, Slivova V, Jiang J, Sliva D. Biochem Biophys Res Commun. 2005 Apr 29;330(1):46-52.

Ganoderma lucidum (G. lucidum) is a popular medicinal mushroom that has been used as a home remedy for the general promotion of health and longevity in East Asia. The dried powder of G. lucidum, which was recommended as a cancer chemotherapy agent in traditional Chinese medicine, is currently popularly used worldwide in the form of dietary supplements. We have previously demonstrated that G. lucidum induces apoptosis, inhibits cell proliferation, and suppresses cell migration of highly invasive human prostate cancer cells PC-3. However, the molecular mechanism(s) responsible for the inhibitory effects of G. lucidum on the prostate cancer cells has not been fully elucidated. In the present study, we examined the effect of G. lucidum on angiogenesis related to prostate cancer. We found that G. lucidum inhibits the early event in angiogenesis, capillary morphogenesis of the human aortic endothelial cells. These effects are caused by the inhibition of constitutively active AP-1 in prostate cancer cells, resulting in the down-regulation of secretion of VEGF and TGF-beta1 from PC-3 cells. Thus, G. lucidum modulates the phosphorylation of Erk1/2 and Akt kinases in PC-3 cells, which in turn inhibits the activity of AP-1. In summary, our results suggest that G. lucidum inhibits prostate cancer-dependent angiogenesis by modulating MAPK and Akt signaling and could have potential therapeutic use for the treatment of prostate cancer.

Ganoderma lucidum inhibits proliferation and induces apoptosis in human prostate cancer cells PC-3.

Jiang J, Slivova V, Valachovicova T, Harvey K, Sliva D. Int J Oncol. 2004 May;24(5):1093-9.

Ganoderma lucidum (Reishi), an oriental medical mushroom, has been widely used in Asian countries for centuries to prevent or treat different diseases, including cancer. However, the mechanism(s) responsible for the effects of Ganoderma lucidum on cancer cells remain to be elucidated. We have previously demonstrated that Ganoderma lucidum down-regulated the expression of NF-kappaB-regulated urokinase plasminogen activator (uPA) and uPA receptor (uPAR), which resulted in suppression of cell migration of highly invasive human breast and prostate cancer cells. In this study, we investigated the effects of Ganoderma lucidum on cell proliferation, cell cycle, and apoptosis in human prostate cancer cells PC-3. Our data demonstrate that Ganoderma lucidum inhibits cell proliferation in a dose- and time-dependent manner by the down-regulation of expression of cyclin B and Cdc2 and by the up-regulation of p21 expression. The inhibition of cell growth was also demonstrated by cell cycle arrest at G2/M phase. Furthermore, Ganoderma lucidum induced apoptosis of PC-3 cells with a slight decrease in the expression of NF-kappaB-regulated Bcl-2 and Bcl-xl. However, the expression of proapoptotic Bax protein was markedly up regulated, resulting in the enhancement of the ratio of Bax/Bcl-2 and Bax/Bcl-xl. Thus, Ganoderma lucidum exerts its effect on cancer cells by multiple mechanisms and may have potential therapeutic use for the prevention and treatment of cancer.

Ganoderma lucidum suppresses motility of highly invasive breast and prostate cancer cells.

Sliva D, Labarrere C, Slivova V, Sedlak M, Lloyd FP Jr, Ho NW. Biochem Biophys Res Commun. 2002 Nov 8;298(4):603-12.

A dried powder from basidiomycetous fungi, Ganoderma lucidum, has been used in East Asia in therapies for several different diseases, including cancer. However, the molecular mechanisms involved in the biological actions of Ganoderma are not well understood. We have recently demonstrated that phosphatidylinositol 3-kinase (PI 3-kinase) and nuclear factor-kappaB (NF-kappaB) regulate motility of highly invasive human breast cancer cells by the secretion of urokinase-type plasminogen activator (uPA). In this study, we investigated the effect of G. lucidum on highly invasive breast and prostate cancer cells. Here we show that spores or dried fruiting body of G. lucidum inhibit constitutively active transcription factors AP-1 and NF-kappaB in breast MDA-MB-231 and prostate PC-3 cancer cells. Furthermore, Ganoderma inhibition of expression of uPA and uPA receptor (uPAR), as well secretion of uPA, resulted in the suppression of the migration of MDA-MB-231 and PC-3 cells. Our data suggest that spores and unpurified fruiting body of G. lucidum inhibit invasion of breast and prostate cancer cells by a common mechanism and could have potential therapeutic use for cancer treatment.

Saw palmetto/Ju Zong Lu

Saw Palmetto induces growth arrest and apoptosis of androgen-dependent prostate cancer LNCaP cells via inactivation of STAT 3 and androgen receptor signaling.

Yang Y, Ikezoe T, Zheng Z, Taguchi H, Koeffler HP, Zhu WG. Int J Oncol. 2007 Sep;31(3):593-600.

PC-SPES is an eight-herb mixture that has an activity against prostate cancer. Recently, we purified Saw Palmetto (Serenoa repens) from PC-SPES and found that Saw Palmetto induced growth arrest of prostate cancer LNCaP, DU145, and PC3 cells with ED50s of approximately 2.0, 2.6, and 3.3 microl/ml, respectively, as measured by mitochondrial-dependent conversion of the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Saw Palmetto induced apoptosis of LNCaP cells in a time- and dose-dependent manner as measured by TUNEL assays. Also, Saw Palmetto increased the expression of p21waf1 and p53 protein in LNCaP cells. In addition, we found that Saw Palmetto down-regulated DHT- or IL-6-induced expression of prostate specific antigen in conjunction with down-regulation of the level of androgen receptor in the nucleus as measured by Western blot analysis. Moreover, Saw Palmetto down-regulated the IL-6-induced level of the phosphorylated form of STAT 3 in LNCaP cells. Furthermore, Saw Palmetto inhibited the growth of LNCaP cells present as tumor xenografts in BALB/c nude mice without adverse effect. These results indicate that Saw Palmetto might be useful for the treatment of individuals with prostate cancer.

Effects of dietary saw palmetto on the prostate of transgenic adenocarcinoma of the mouse prostate model (TRAMP).

Wadsworth TL, Worstell TR, Greenberg NM, Roselli CE. Prostate. 2007 May 1;67(6):661-73.

BACKGROUND: Several of the proposed mechanisms for the actions of the liposterolic extract of saw palmetto (SPE) are exerted on known risk factors for prostate cancer (CaP). This study investigated whether SPE could prevent the progression of CaP in a transgenic adenocarcinoma of the mouse prostate (TRAMP) model. METHODS: Two different doses of SPE designed to deliver 50 mg/kg/day SPE and 300 mg/kg/day SPE were administered in a custom diet to TRAMP mice for 12 or 24 weeks. Body and organ weights were used to evaluate toxicity, and radioimmunoassay was used to measure plasma and tissue androgen levels to monitor effects of SPE on 5alpha reductase activity. Prostate tissues were evaluated histologically to determine the effect of treatment on tumor grade, cell proliferation, and apoptosis. RESULTS: Treatment with 300 mg/kg/day SPE from 4 to 24 weeks of age significantly reduced the concentration of 5alpha-dihydrotestosterone (DHT) in the prostate and resulted in a significant increase in apoptosis and significant decrease in pathological tumor grade and frank tumor incidence. CONCLUSIONS: Dietary supplementation with SPE may be effective in controlling CaP tumorigenesis. SPE suppression of prostatic DHT levels lends support to the hypothesis that inhibition of the enzyme 5alpha-reductase is a mechanism of action of this substance.

Effects of homeopathic preparations on human prostate cancer growth in cellular and animal models.

MacLaughlin BW, Gutsmuths B, Pretner E, Jonas WB, Ives J, Kulawardane DV, Amri H. Integr Cancer Ther. 2006 Dec;5(4):362-72.

The use of dietary supplements for various ailments enjoys unprecedented popularity. As part of this trend, Sabal serrulata (saw palmetto) constitutes the complementary treatment of choice with regard to prostate health. In homeopathy, Sabal serrulata is commonly prescribed for prostate problems ranging from benign prostatic hyperplasia to prostate cancer. The authors' work assessed the antiproliferative effects of homeopathic preparations of Sabal serrulata, Thuja occidentalis, and Conium maculatum, in vivo, on nude mouse xenografts, and in vitro, on PC-3 and DU-145 human prostate cancer as well as MDA-MB-231 human breast cancer cell lines. Treatment with Sabal serrulata in vitro resulted in a 33% decrease of PC-3 cell proliferation at 72 hours and a 23% reduction of DU-145 cell proliferation at 24 hours (P<.01). The difference in reduction is likely due to the specific doubling time of each cell line. No effect was observed on MDA-MB-231 human breast cancer cells. Thuja occidentalis and Conium maculatum did not have any effect on human prostate cancer cell proliferation. In vivo, prostate tumor xenograft size was significantly reduced in Sabal serrulata-treated mice compared to untreated controls (P=.012). No effect was observed on breast tumor growth. Our study clearly demonstrates a biologic response to homeopathic treatment as manifested by cell proliferation and tumor growth. This biologic effect was (i)significantly stronger to Sabal serrulata than to controls and (ii)specific to human prostate cancer. Sabal serrulata should thus be further investigated as a specific homeopathic remedy for prostate pathology.

Berberine

Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells.

Mantena SK, Sharma SD, Katiyar SK. Mol Cancer Ther. 2006 Feb;5(2):296-308.

Berberine, a naturally occurring isoquinoline alkaloid, has been shown to possess anti-inflammatory and antitumor properties in some in vitro systems. Here, we report that in vitro treatment of androgen-insensitive (DU145 and PC-3) and androgen-sensitive (LNCaP) prostate cancer cells with berberine inhibited cell proliferation and induced cell death in a dose-dependent (10-100 micromol/L) and time-dependent (24-72 hours) manner. Treatment of nonneoplastic human prostate epithelial cells (PWR-1E) with berberine under identical conditions did not significantly affect their viability. The berberine-induced inhibition of proliferation of DU145, PC-3, and LNCaP cells was associated with G1-phase arrest, which in DU145 cells was associated with inhibition of expression of cyclins D1, D2, and E and cyclin-dependent kinase (Cdk) 2, Cdk4, and Cdk6 proteins, increased expression of the Cdk inhibitory proteins (Cip1/p21 and Kip1/p27), and enhanced binding of Cdk inhibitors to Cdk. Berberine also significantly (P < 0.05-0.001) enhanced apoptosis of DU145 and LNCaP cells with induction of a higher ratio of Bax/Bcl-2 proteins, disruption of mitochondrial membrane potential, and activation of caspase-9, caspase-3, and poly(ADP-ribose) polymerase. Pretreatment with the pan-caspase inhibitor z-VAD-fmk partially, but significantly, blocked the berberine-induced apoptosis, as also confirmed by the comet assay analysis of DNA fragmentation, suggesting that berberine-induced apoptosis of human prostate cancer cells is mediated primarily through the caspase-dependent pathway. The effectiveness of berberine in checking the growth of androgen-insensitive, as well as androgen-sensitive, prostate cancer cells without affecting the growth of normal prostate epithelial cells indicates that it may be a promising candidate for prostate cancer therapy.

Inhibition of tumor-induced angiogenesis and matrix-metalloproteinase expression in confrontation cultures of embryoid bodies and tumor spheroids by plant ingredients used in traditional chinese medicine.

Wartenberg M, Budde P, De Marees M, Grunheck F, Tsang SY, Huang Y, Chen ZY, Hescheler J & Sauer H. Lab Invest. 2003 Jan;83(1):87-98.

Tumor-induced angiogenesis is a prerequisite for excessive tumor growth. Blood vessels invade the tumor tissue after degradation of the extracellular matrix scaffold by matrix metalloproteinases (MMPs). Inhibition of MMPs has been therefore suggested to be a useful tool to abolish neoangiogenesis of solid tumors. In the present study, antioxidative plant ingredients used in traditional Chinese medicine were investigated for their capacity to down-regulate MMP expression and to inhibit angiogenesis in embryonic stem cell-derived embryoid bodies and tumor-induced angiogenesis in confrontation cultures consisting of embryoid bodies and multicellular DU-145 prostate tumor spheroids. Embryoid bodies transiently expressed MMP-1, MMP-2, and MMP-9 during the time of differentiation of capillary-like structures. In confrontation cultures, MMP expression was increased compared with control tumor spheroids and embryoid bodies cultivated separately. The increased expression of MMPs in confrontation cultures was a result of elevated levels of reactive oxygen species (ROS) upon confrontation culture and was totally abolished in the presence of the free radical scavenger vitamin E. Incubation of embryoid bodies with baicalein, epicatechin, berberine, and acteoside, which are herbal ingredients used in traditional Chinese medicine, significantly inhibited angiogenesis in embryoid bodies and decreased intracellular ROS levels. Tumor-induced angiogenesis in confrontation cultures was totally abolished in the presence of the free radical scavenger vitamin E. Because herbal ingredients down-regulated MMP expression, we conclude that ROS generated during confrontation culture induce the expression of MMPs that are necessary for endothelial cell invasion into the tumor tissue.