| Ribonuclease inhititor(RI) is an acidic cytoplasmic glycoprotein, its molecular weight is 50KDa. RI can inhitit the activity of ribonuclease A (RNaseA) by tight combination with it. Angiogenin(Ang)is produced and secreted by tumor cells and normal cells, it has a robust capacity to induce blood vessel formation, its molecular mass is 14.4KDa.Ang is a member of RNase superfamily. It is expressed widely in many kinds of cell and tissue. It can express both in cancer tissues and normal tissues. The wide expression of Ang suggests that it is restrictively controlled in the body. Ang is 35% identical with human pancreatic RNase, X-ray analysis showed that Ang had a very similar space structure. They can both form tight complex with RI. Because RI can form the tight complex with Ang, so it can inhibit the activity of Ang. Previous study indicated that RI can effectively inhibit the activity of Ang to inhibit the blood vessel formation to slow down the growth and metastasis of tumor.Ovarian cancer is a kind of parenchymatous tumor, there are high density capillary vessels in tumor interstitial substance, which is related to the growth and metastasis of tumor. In initial phase of the malignant tumor cell growth, there isn't autospecific blood vascular system, the nutrition is provided by simple diffusion,there is no metastasis. When the gross tumor volume upto 1-2mm3, New vessels are necessary to provide nutrition and oxygen for the development of tumors and their metastatic dissemination. So anti-angiogenesis will be an efficient method in controlling ovarian cancer.Presently, many angiogenetic inhibitors have been used to ovarian cancer, but the effect of gene therapy are more signigicant, which transfected certain gene to tumor cells, ensure the gene can be continue expressed, in order to control vascularization persistently. Gene therapy emergy us a promising perspective for ovarian cancer.Objective: To explore the transfection and expression of RI gene on human ovarian cancer cell HO-8910 and observe whether the growth of tumor cells could be inhibited.Methods:1.The recombination vector carrying human ribonuclease inhibitor gene plncx-ri was transfected in HO-8910 cell using the lipofectamine -mediated method.2.After selection in the presence of G418 in culture medium, cell colons were obtained.3.The HO-8910 plncx-ri cell line with a stably high expression of RI was identified by RT-PCR and immunocytochemistry.4.Observe cellular morphology Cell counting, MTT and FCM were used to observe the effect of proliferation to ovarian cancer cell.Result:1.The gene plncx-ri was transfected in HO-8910 cell using the lipofectamine 2000, with 400ug/mlG418 the positive cell colons were obtained.2.The plncx-ri on chromosomes was found by RT-PCR.3.The positive RI gene expression of HO-8910 cells before or after transfection were confirmed by immunocytochemistry.4.The growth of tranfected HO-plncx-ri cells were slower. After transfected the growth inhibiting of cells after 24h,48h,72h,96h,is increased distinctly. The HO-plncx-ri cells of G1 stage increased, P<0.05,and the cells of S stage were reduced, P<0.05.Conclusion: The RI gene could be transfect into human ovarian cancer cell HO-8910 and the expression quantity was high, the growth of tumor cells could be inhibited.
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