9. Collaborative Studies of GCP
Proceedings of AHCC Research Association 8th Symposium
1. GCP: A Genistein Combined Polysaccharide with anti-angiogenesis and anti-tumor activities in vitro and in vivo
Lan YUAN, Takehito MIURA, Buxiang SUN, Mayumi YOSHITA, Hajime FUJII, Kenichi KOSUNA Laboratory of Biochemstry, Amino Up Chemical, Co. Ltd., Sapporo 004-0839, Japan.
Genistein Combined Polysaccharides (GCP) were detected for their anti-angiogenensis activity by in vitro, in vivo and ex ovo methods in this study. GCP inhibited 50% of mouse brain vascular endothelial cell LE-1 on their proliferation in vitro. In three-dimensional collagen gels angiogenesis in vitro assay, GCP inhibited the new blood vessel formation. The rate of growth of microvessels from the perimeter of rat aortic rings embedded in fibrin gel was measured. GCP inhibited the 96% of growth of microvessels as compared with PBS treated group. Both of the inhibitions were much higher than that of purified genistein. A chamber angiogenesis assay in vivo was performed by use of mouse colon cancer cells Colon-26 inoculated into the back of the mouse subcutaneously. The results showed that tumor cells in the chamber induced new blood vessels (angiogenesis) were significantly inhibited in GCP-treated mouse. GCP also inhibited chick embryo chorioallantoic membrane (CAM) angiogenesis ex ovo by dose-dependent manner and more effective than commercial isoflavone. For the anti-tumor activity studies, GCP inhibited several kinds of tumor cells by induction of apoptosis. GCP significantly inhibited the tumor growth in both melanoma and sarcoma tumor bearing animal models. These results strongly suggested that GCP is an effective antiangiogenic agent and might be used in cancer treatment.
Key Words: isoflavone, genistein, polysaccharide, tumor, angiogenesis.
2. Studies on the inhibition of angiogenesis by GCP
Paul F. Davis, Bei Xu.
Bioactivity Investigation Group, Wellington School of Medicine, P.O. Box 7343, Wellington South, New Zealand
Cancerous tumors need an adequate blood supply in order to grow. So inhibiting the growth of new blood vessels will slow the growth of cancers. As well as the isoflavone, genistein, GCP also contains sugars, polysaccharides and amino acids. So it is predicted that GCP will inhibit the development of new blood vessels (angiogenesis) and so be useful in the treatment of solid cancers. When mixed with water, some of the GCP dissolved. Testing of this solution at different concentrations showed that it was anti-angiogenic. This suggested that there was a constituent other than genistein that was responsible for this inhibition.. When GCP is dissolved in dimethylsulfoxide (DMSO) there was greater than 90% inhibition of angiogenesis at 1.5mg/ml. This was the equivalent of 10�g/ml of genistein. In order to investigate this further, GCP was extracted with ethanol. About 36.5% of the GCP and more than 95% of the genistein dissolved in this. However when this was tested in the angiogenic assay, it could not be evaluated as the fibrin gel matrix dissolved. This will be discussed further. The ethanol-insoluble fraction was extracted with water. This solution which contained 48% of the GCP (mostly sugars and amino acids) but only 1.9% of the genistein as strongly anti-angiogenic with 50% inhibition of 25�g/ml.
These results and other observations confirmed that GCP is potently anti-angiogenic with both the genistein and the water-soluble constituents both contributing to its anti-angiogenic properties.
3. Various effects of the co-administration of AHCC and GCP in vitro and in vivo
Buxiang Sun, Ph.D., Amino Up Chemical Co. LTD.;
Xubao Shi, Ph.D., Research Scientist, Medical Center of UC Davis;
Ralph deVere White, MD., Director of Cancer Center, Medical Center of UC Davis;
Robert Hackman, Ph.D., Research Professor, Nutrition Department of UC DavisStudies have demonstrated that GCP has the anti-agiogensis, suggesting its role in anti-tumor and AHCC can increase the immunoreactivity. Here, we investigated their synergistic anti-tumor effects in vitro and in vivo. Six human cancer cell lines and two mouse carcinoma cell lines were used in the studies. Of eight cell lines, four stem from prostate (LNCaP, PC3, DU145 and TSU-prl), one from bladder (T24), one from bone (Saos-2), one from lung (3LL), and one from colon (Colon 26). MTT assay was used to measure the inhibitory effects on cell growth. It was found that both GCP and GCP+AHCC could inhibit the growth of all eight cell lines tested in cell growth. It was found that both GCP and GCP+AHCC could inhibit the growth of all eight cell lines tested in dose-dependent manners, but AHCC did not show obvious inhibitory effect on these cell lines except Colon 26. We examined the expression levels of VEGF using ELISA, and found that GCP can down-regulate the VEGF expression depending on the cell lines. We observed that GCP and AHCC induced apoptosis in several cell lines. PC3 cells were treated with these reagents, and Western blotting analysis of PARP protein and TUNEL assay were used to examine the apoptosis of tumor cells. GCP + AHCC induced obvious apoptotic death when compared to GCP and AHCC alone. Furthermore, the similar effect was observed in T24 and 3LL cells. In addition, GCP+AHCC was also found to up-regulate p21 and down-regulate VEGF in PC-3 cells. GCP+AHCC shows a synergistic effect on inhibition of tumor growth in animals. Nude mice bearing PC3 were separately treated with GCP+AHCC, GCP and AHCC. Although the inhibitory effects on the tumor growth were observed in all three groups, the inhibition in GCP+AHCC group was much more obvious than in the other two groups. The synergistic effect on tumor growth inhibition was also observed in 3LL cells and in Sarcoma 180 cells. When the treatment was stopped in AHCC group and GCP+AHCC group, the PC3 tumors in both groups showed the same growth was stopped in AHCC group and GCP+AHCC group, the PC3 tumors in both groups showed the same growth was stopped in AHCC group and GCP+AHCC group, the PC3 tumors in both groups showed the same growth was stopped in AHCC group and GCP+AHCC group, the PC3 tumors in both groups showed the same growth rate as in the control group untreated. In summary, GCP and AHCC show anti-tumor effects by inducing apoptosis and/or antiagogenesis in these cell lines tested. The anti-tumor effects can be increased by a combination administration of GCP and AHCC.
4. A Mixture of Basidiomycetes Polysaccharide and Genistein (GCP) Inhibits Proliferation and Induces Apoptosis in Human Prostate Cancer Cells in vitro and in vivo
R. Buttyan, Aaron E. Katz, Y. Cao, T. Dorai, C. Olsson
Department of Urology and Pathology, Columbia University, College of Physicians and Surgeons, New York, New York 10032.In our studies at Columbia University in New York, we have performed several experiments in the laboratory GCP. It is clear tat GCP can have a dramatic effect on prostate cancer cells. GCP can cause prostatic cancer cells to stop growing and dividing. This finding as been duplicated in both cell culture and animal models. While the exact mechanism of actions is unknown, we have found that GCP can block an important regulator of cell cycle function, known as p27. In addition, when cancer cells are exposed to GCP, they undergo a mechanism of cell death known as apoptosis. The death of these cells can occur early, often within a few yours after exposure to GCP. In animals that were implanted with prostate tumors, GCP caused significant regression of these tumors.
GCP may have a number of potential roles in fighting cancer, as well as in cancer prevention. This is an exciting new compound from Japan and may play a significant role in the management of cancer here in the United States.
Other Proceedings
- Yuan L. Inhibition of tumor proliferation and angiogenesis by genistein Combined polysaccharide in vitro and in vivo. The First International China Angiogenesis Program, Molecular Strategies and clinical development. November 1999, Beijing, China.
- Miura T, and Yuan L. et al. Anti-tumor and anti-angiogenesis effects of soybean isoflavone aglycones and extracts from cultural Basidiomycetes mycelium. Nippon Nogeikagau Kaishi. 74:68, 2000.
Home | | News | Poster Presentation
GCPresearch.com P.O. Box 311 Rye, NY 10580
Tel: (914) 251-0255 E-fax: (775) 599-7918
Email: [email protected]