Study On Effect Of Antimony Trioxide On The Proliferation Inhibition In SKOV3 Cell Of Ovarian Carcinoma

Objective: Ovarian cancer is one of the most lethal tumor of the female genital tract and is seriously threatening female health. Lacking of early effective diagnosis methods and treatment, 70 percent of patients with ovarian cancer are diagnosed as late-stage's. The conventional principles of treatment are surgery and chemotherapy. 5-year survival is not obviously increased. It is necessary to find a new effective anti-neoplastic drug to improve the prognosis of the patients with ovarian cancer. Antimony-als and its compounds have been widely applied to mankind for a long history, primarily employed as old anti-parasite drug. People bolted synthesis anticancer compound to discover mang kinds antimony compounds to have anticancer activity. But the lucubrate had not been undertaken. Our recent studies showed people also renewed the anticancer activity of the antimony. Related scholars found that micromole concentration of the antimony could induce growth inhibition in acute promyelocytic leukemia (APL) cell lines, malignany lymphocyte cell growth, but no infulunce in norm-lymmphocyte cell. Its mechanism induced apoptosis. Yu Wen Qiang et al found trivalent antimony-als could effectly induced acute promyelocytic leukemia (APL) cell lines to apoptosis, showing trivalent antimony-als had a widely prospective in the therapy of anti-cancer. This experiment researched the effect on the ovarian cancer cell SKOV3 of Sb2O3, and it's cell proliferation inhibition and apoptosis function provided a new though and method for the study of anti-tumor.Methods: 1 Sb2O3 was dissolved by dilute hydrochloric acid with the concentration of 1.1mmoL/L, and preserved no more than 3 days at 4℃closed-light. Stired at least 30 min's before it was used. Diluted by RPMI-1640 contained 10% FCS into 6.25~25μmol/L. 2 SKOV3 ovarian cancer cells were seeded in 96-well microtiter plates and culture bottles in the same concentration and maintained in RPMI-1640 contained 10% FCS in 37.℃, 5% CO2 and allowed to adhere for 24 hours . Then cells were dvided into four groups: untreated control group; treated groups in different concentration. Each group cells were transfected with different concentration Sb2O3 or equaled culture liquid for 24,48 and 72 hours. 3 Cell proliferation inhibition was measured by MTT assay. 4 Morphological change of apoptotic cells were observed by Light microscope, Electric microscope and Giemsa staining. 5 Test the DNA Ladder extract the DNA and further test the characteristic DNA degradtion with 1.5% agarose gel electrophoresis. 6 The Flow Cytometry Analysis made a quantitative analysis with the flow cytometry on the apoptosis rate of the Ovarian cancer cells, the specificity of the cell cycle, and the expression of bcl-2 and bax protein.Results: 1 MTT assay showed that cell proliferation was significantly inhibited by Sb2O3 in time and dose dependent manner. Ovarian cancer cell SKOV3 was effect by medical serum for 24,48 or 72 hours. 2 Ovarian cancer cell SKOV3 treated by Sb2O3 showed characteristic morphological features and super-microstructural changes of apoptosis were observed by means of light microscope, Giemsa staining and electric microscope, including cell shrinkage, cytoplasm concentration, nclius forming outwardly acute angle promineney, chromatin concentrating on karyotheca or forming meniscusnuclear condensation,electric density increasing, nuclear fragmentation. but no formation of multiplicity typical apoptotic bodies. 3 DNA agarose gel electrophoresis showed characteristic"DNA ladder"pattern by Antimony trioxide treatments. Gradient strap became obivious with increasing of the concentration. But DNA in the control group didn't show this change. 4 Flow Cytometry Apoptotic rate was different between trial groups and control group (P<0.01), Cell cycle analyzed was singnificant difference, cells in S and G1/G0 phase were reduced and in G2 /M phase were enhanced in trial groups compared to control group (P<0.01). the effect was in dose-dependent manner. 5 Expressive amount of bcl-2 and bax protein the indirect immum-ofluorescence of the flow cytometry showed the expression of bcl-2 was inhibited and the expression of bax was up-regulated. It indicates that Antimony trioxide induced apoptosis of ovarian cancer cells that might attribute to the down-regulation of anti-apoptotic ihhibitors bcl-2 and the up- regulation of anti-apoptotic promotion bax.Conclusion: 1 Sb2O3 has the function of proliferation inhibition of ovarian cancer cell, And this effect is in dose and time dependent manner. 2 Antimony trioxide can effectively induce apoptosis of SKOV3 cell , induced-apoptosis may be one of its mechanism of inhibition cell growth. 3 This apoptosis may be mediated by down expression of bcl-2 and up expression of bax. 4 This research provides a new thought of the anti-tumor therapy both in ovarian cancer and Antimony trioxide. Antimony trioxide may be a promising apoptosis-inducer in ovaria cancer therapy.