| Objective: To detected the expression of Livin in human ovarian cancer parent cells SKOV3 and Cisplatin resisitant cells SKOV3/CDDP. To explore the changes of the expression of the Livin mRNA,the reproductive activity and the apoptosis of SKOV3/CDDP cells after transfection .To discuss the possibility of Livin as a therapeutic target and strategies on reversing the drug resistance on SKOV3/CDDP cellsMethods:1 Resuscitate and transfer of culture the SKOV-3 and SKOV3/CDDP cell lines,observed the two cell lines by inverted microscope.2 Detect the expression of the two cell lines by RT - PCR3 Transfect the Livin-RNAi plasmid into SKOV3/CDDP cells by liposome , silent the livin gene and detect the expression of livin mRNA in control group,non-specific transfect group and specific transfect group by RT-PCR .4 MTT assay was performed to measure the reproductive activity of the SKOV3/CDDP cells after transfection.5 the apoptosis of SKOV3/CDDPcells was assessed by FCM after transfectionResults:1 The observation of the two cells by inverted microscope:1.1 SKOV-3cells in inverted microscope:the culture solution was clear and the cells seened to be fusiform shape,concentrated growth, the glossiness and diopter was well.1.2 SKOV3/CDDPcells in inverted microscope:the culture solution is clear , and the cells seens to be polygon , concentrated growth , the glossiness and diopter is good. 2 RT - PCR showed livin mRNA expressioned both in SKOV3/CDDP cells and SKOV- 3 cells, The ratio of Livin mRNA in SKOV3/CDDP cells was 50.88%±1.37compareed with 23.66±0.91% in SKOV-3 cells .The difference was significant(P<0.05),It indicated that the expression of Livin mRNA in SKOV/CDDP cells is higher than in SKOV- 3 cells.3 Obeserved the three group cells by inverted microscope after transfect for 48hours,the control group cells and non-specific group cells were overlapping growth,the specific group cells density was lower,glossness comes down,part of the transfected groups cells were express green fluorescence by fluorescence microscope. the rate of expressing green flourscencn cells was>70%,the transfection was successful.4 RT - PCR showed 48h after Livin targeted microRNA transfect,Livin mRNA expressioned in control group,non-specifc tranfect group and specific transfect group.the The ratio of Livin mRNA in specific transfect group was 13.83±0.47%, but the control group and non-specific transfect group were 50.47±0.46%,49.71±0.57%,The difference in specific transfect group and the control group and non-specific transfect group were significant(P<0.05), it showed that after transfection the Livin mRNA expression was weakened, the RNA interference for SKOV3/ CDDP was significant. The micRNA interference for SKOV3/CDDP is feasible.5 MTT showed that the OD of specific transfect group after 24h of transfect was 0.474±0.04,but the non-specific transfect group and the control group was 50.47±0.46%,49.71±0.57%,compare with the speci- fic transfect group,the difference was significant(P<0.05).48h after transfected the OD in the three groups were 0.353±0.05, 0.612±0.07,0.634±0.05,and 72h after transfected ,the OD in the three groups were 0.410±0.020, 802±0.07,0.780±0.06. compare with the specific transfect group,the difference also was significant (P<0.05).it showed that after transfected the reproductive activity of the SKOV3/CDDP cells were been inhibited.6 FCW showed the ratio of apoptosis in three group after 48h of transfect:the the specific transfect group was14.86±0.37,and the non-specific transfect group and the control group was 1.15±0.98, 1.62±0.34, ,the ratio of apoptosis in non-specific transfect group and the control group was significant compare with the specific transfect group (P<0.05).it indicated that the silencing of Livin can induce apoptosis of SKOV3/CDDPcellsConclusion:Our research indicated that high expression of Livin gene has close relationship with drug resistance. Livin targeted microRNA transfect can weaken the expression of Livin mRNA. After transfect, the proliferation of the cells was been inhibited and the apoptosis was been induced. Livin gene was to be a possible therapeutic target and strategies on reversing the drug resistance on SKOV3/CDDP cells.
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