The Effects Of Livin Targeted MicroRNA On Hunman Ovarian Carcinoma Drug-resistance Cell In Vitro

Objective: Transfected hunman ovarian carcinoma drug-resistance cells with microRNA against Livin , to study the effect of downregulation of Livin on cisplatin resistance of hunman ovarian carcinoma drug-resistance cells. Diseusses the relationship of Livin and the drug resistance of ovarian carcinoma.Methods:1 Human ovarian cancer cell SKOV3 and SKOV3/CDDP resuscitative and culture,cells morphological were observed by inverted microscope.2 Check the IC50 of the drug-resistant ovarian cancer cell SKOV3/CDDP to SKOV3 through MTT,and calculate resistant index.3 Designed and synthesised the Livin targeted microRNA and linked to plasmid Vector. Transfect the SKOV3/CDDP cells beforehand by the liposome. 48h after that,The green fluorescence cell and the transfection efficiency were observed.4 The proliferation ability of ovarian carcinoma cells was test by MTT experiment.The apoptosis percent of ovarian carcinoma cells were assessed with flowcytometry.Results:1 The resistant index of SKOV3/DDP is 3.29,which could be used in the test.2 The observation by fluorescence microscope:48h after transfect,the cells which expression green fluorescnce were 70-80 percent in the transfected group cells .The transfection was successful. 72h after transfect,the cells which expression green fluorescnce begin to reduce.120h after transfect,the cells which expression green fluorescnce is seldom.So my study have been done in 72 hours after transfect. 3 The result which detdcted by MTT show:In the cisplatin of 1ug/ml concentration,the absorbances of the transfected group are 0.56±0.060 at 12 h,0.69±0.056 at 24h and 1.04±0.200 at 48h. In the cisplatin of 5ug/ml concentration,the absorbances of the transfected group are 0.40±0.027 at 12 hours, 0.50±0.002 at 24 hours and 0.700±0.062 at 48 hours. In the cisplatin of 10ug/ml concentration,the absorbances of the transfected group are 0.31±0.033 at 12h,0.38±0.031 at 24h and 0.56±0.074 at 48h.This is show:adding cisplatin of different concentration and in different times ,the cell inerease inhibition is proved to be improved in the transfected group by MTT(P<0.05). (NO significant difference in apoptosis rate is founded between the negative group and control group.)4 Flowcytometry results show: In the cisplatin of 1ug/ml concentration, the apoptosis of the transfected group is 24±3.296% at 24h, the negative group is 8.03±0.354 %, the control group is 7.11±0.644%.This is show:that the apoptosis is obviously increased in the transfected group (P<0.05) . (NO significant difference in apoptosis rate is founded between the negative group and control group.)Conclusion:Livin targeted microRNA transfect human ovarian cancer cell SKOV3/CDDP , can weak the tolerance of cancer cells to cisplatin. It also suggested that Livin might be involved in cisplain-resistance of ovarian cancer cells.The experment is onlypreliminary researche.If we want to get a irrefutably conclusion,we should do the intracorpopal study.