The Mechanisms Of Study On The Derivative Of SC Anti-tumor

The ocean is regarded as the origin of life evolution, and stores a lot of Comprehensive Ocean Creature Resources. With the understanding of the ocean resources and development of modern biological technology applying in the field of pharmacological research of ocean drugs, ascidians (also know as sea squirts), became hot for chemist and pharmacologist in discovering the novel ocean drugs. Recently, some scientists discovered a lot of effective components from ascidiacea with anti-tumor, anti-virus, antimicrobial, immuno-regulation and biocatalyst activity,especially anti-tumor components, such as alkaloid, peptide, Polyether Marine, Ganodermenono and organ-sulfur-compound. During the period of 9th and 10th"five-year plan", we did a lot of researches on the anti-tumor and anti-virus mechanisms of ascidiacea, and also separated and purified some kinds of effective components. Styela clava (SC, 5α- 8α- cyclicobioxygen- 24- bimethyl- 6- vinyl- 3β- cholesterol) have been demonstrated to function as anti-HBV, anti-tumor and immuno-regulatary effects. Base on the structure of SC, we made a serial of modification and careful screen to achieve a derivative of SC- 002,which is also confirmed to have effective anti-tumor activity.MTS assay was used to assess the proliferation inhibition effects of 002 on HepG2, A549 and NIH3T3 cell lines in vitro. Flow cytometry and gel electrophoresis DNA Ladder.was used to observe the influences of 002 on the apoptosis of HepG2,A549,NIH3T3. The data indicated that 002 significantly inhibited the proliferation of HepG2 and A549 cells by 73.67% and 62.40% respectively in a dose-dependent manner,. In addition,002 can also induced apoptosis on HepG2 and A549 but not NIH3T3.Moreover, we also investigate the anti-tumor activity of 002 in vivo in Balb/c mice. The changes of liver, spleen and weight were used as markers to assess the anti-tumor activity of 002. Balb/c mice were respectively inoculated with S180 subcutaneously and H22 intraperitoneally, and simultaneously injected,with 002 in the abdomen at high dose (32 mg/kg)and low dose(16 mg/kg). CTX and corn oil solvent served as controls. The data showed that the high and low doses of 002 can inhibit effectively S180 tumor growth by 54.58% and 44.33%, and prolong the survival days of experimental mice by 51.55% and 37.11% respectively. The experiments of chicken embryo chorioallantoic membrane model in anti-angiogenesis showed that 002 could inhibit the growth of the new blood vessles in certain dose.We also detected the concentration of VEGF in mouse serum and Caspase-3,8,9 in solid tumors with ELISA ( enzyme-linked immunosorbent experiment). We found that 002 could reduce VEGF production but not influence the activity of Caspase-3,8,9. Further analysis indicated that 002 could significantly increase the expression level of P53 and reduce the expression level of VEGF and Bcl-2, but not that of P21. These data indicated that 002 could fight tumor by promoting the high expression of P53, and inducing tumor cell apoptosis through reducing the expression level of VEGF and Bcl-2.