Survivin Expression In Ovarian Carcinoma And The Effect Of Antisurvivin Oligonucleotides On Ovarian Carcinoma Cell Development And Sensitivity To Chemotherapy

Objectives: (l)To investigate the possible relationship between expression of survivin and molecular mechanisms in patients with ovarian carcinoma(OC); (2)To study the effect of antisene oligonucleotides(ASODN) and DDP targeting survivin on SKOV3 cells and the possibility of ASODN improving the sensativity of SKOV3 to DDP.Methods: Survivin expression was studied in 10 cases of normal ovaries,21 cases of ovarian bordline tumors,29 cases of ovarian germ cell tumors,38 cases of ovarian serous cystadenomas by means of immunohistochemistry. ASODN was introduced by Lipofectamine^?2000 transfection reagent into SKOV3 cells.The effects of treatment with ASODN alone and/or cisplatin(DDP) therapy on SKOV3 cells growth inhibition were measured by MTT assays and the changes of survivin mRNA was detected by RT-PCR technique.The changes of cell cycle,survivin protein,apoptosis index(AI)were studied by flow cytometry.SPSS 11.0 and STATA 5.0 were made use of for data manipulation and analysis.Results: (l)No survivin protein was expressed in normal ovaries.(2)Survivin protein was found in cytoplasm and/or in nuclear. Survivin presented in 42.9% of cases(9/21) of ovarian bordline tumors.No significant correlation was found between survivin expression and pathologic types or CA125 values(P>0.05).(3) Survivin protein presented in 58.6% of cases(17/29) of ovarian germ cell tumors.lt had no significant correlation with pathologic typesjtumor stage,rymphonode metastasis,AFP values and volume of ascites(P>0.05).(4) Survivin protein was found in 76.3%(29/38) of cases of ovarian serous cystadenomas.With a significant higher expression than that in bordline tumors(P<0.05),With no significant difference with germ cell tumors(P>0.05).Significant correlation was found among survivin expression, tumor stage and grade,lymphonode metastasis(P<0.05).No significant correlation with CA125 values,volume of ascites and whether had tumor cells(P>0.05).(5)ASODN could inhibit SKOV3 cells growth in dose-dependent manner.After 48 hours of transfection,it reached the peak.Neither SODN nor Lipofectamine?2000 affected the tumor cells growth(P>0.05).(6)DDP could inhibit SKOV3 cells growth in dose- and time-dependent manners(P<0.05).After 48 hours of DDP usejcso was over 8ug/ml.(7) Significant higher inhibition was found in the group in which . ASODN as combined with DDP than the group DDP or ASODN was used seperately(P<0.05).After 48 hours of 200nmol/L ASODN combined with DDP use,Ic5o of DDP was only 3ug/ml.That means it was 2.6 times DDP need when DDP used separately than it combined with ASODN.(8)Thetransfection of ASODN in SKOV3 cells resulted in phase Gl arrest,the proportion of G2/M phrase cells and survivin protein decreased,AI increased(P<0.05). (9) Survivin mRNA was much lower in the group with a combination of ASODN with DDP than ASODN group or DDP group(P<0.05).Conclusions: (l)High expression of survivin protein may reveal the pathogenesis of epithelial ovarian carcinoma. So survivin overexpression could be exploited for early diagnostic purpose.(2)The biological function of survivin gene is realized by regulating tumor cells proliferation and apoptosis. (3)The expression levels of survivin protein/mRNA and the proportion of mitotic cells may be decreased and apoptosis AI may be increased in SK0V3 cells after ASODN transfection.(4)ASODN can improve the sensativity of SKOV3 to DDP,hence the less amount of DDP use.