A Study On The Role Of MiR-205 In Proliferation And Invasion Of Cervical Cancer

?Objective?We detected the level of microRNA-205 in the tissue of cervical cancer,cervical intraepithelial neoplasia and normal cervix to analyze its correlation with the main clinical parameters of patients with cervical cancer.And to detect the level of miRNA-205 in vitro and investigate the effect of down-regulation of miRNA-205 on the proliferation,apoptosis and invasion of cervical cancer cells.In order to discuss the possibility of miR-205 as a prognostic indicator and biological treatment of cervical cancer.?Methods? 1.50 speciments of cervical cancer,30 speciments of cervical intraepithelialneoplasia(CIN)and 10 speciments of benign uterine lesions were collectedrandomly.The miR-205 expression level of the tissues above were measured byquantitative RT-PCR and to analyze its expression changes in different sampleabove.2.The clinical parameters of cervical cancer patients were collected,includingtumor size,tumor cell differentiation,FIGO clinical stage,lymph nodemetastasis,cancer cell growth index.The correlation between the miR-205 leveland the clinical parameters were analyzed.3.We studied on the miR-205 inhibitor effect in vitro with HeLa cells.It was usedquantitative RT-PCR to detect the expression of miR-205 in miR-205 inhibitorgroup,inhibitor control group and blank control group of the cells,MTT wasused to detect the cell proliferation,Flow cytometry was used to detect the cellapoptosis,Transwell was used to test the cell invasion.?Results? 1.The expression levels of miR-205 in cervical cancer tissues were significantlyhigher than that in CIN tissues and normal human cervical epithelium(p < 0.001);and miR-205 expression in CIN tissues much higher than normal human cervicalepithelium,the difference was significant of statistics(p <0.001).2.The level of miR-205 in those patients with cervical cancer ?IIb stage,weresignificantly higher than that patients of ?IIa stage.The difference wassignificant of statistics(p=0.002).3.About the patients with cervical cancer ?IIa stage,the level of miR-205 inpatients with lymph node metastasis was significantly higher than that in patientswithout lymph node metastasis.The difference was significant of statistics(p=0.002).4.The expression of miR-205 in patients with tumor maximum diameter >4 cmwas higher than that of patients with tumor diameter ? 4 cm,but the differencewas not significant of statistics(p= 0.06).There was no significant difference imiR-205 levels between exogenous and endogenous tissues(p= 0.15).There wasno significant difference in miR-205 levels among G1,G2 and G3 differentiation(p> 0.05).5.The expression level of miR-205 in HeLa cell line was significantly higher thanthat of human normal cervical epithelial Ect1/E6E7 cells(1.566 folds,p= 0.028).The miR-205 level in HeLa cell line transfected with miR-205 inhibitor was 0.46times,significantly lowerer than that of the inhibitor control group(p= 0.006)and was 0.47 times,significantly lowerer than that of the blank control group(p= 0.001).There was no significant difference in miR-205 level between He Lacells transfected with inhibitor control and the blank control group(p= 0.093).6.The OD value(490nm)of the cells in miR-205 inhibitor group at 24 h,78 h,72 hand 96 h after transfection were significantly lower than the blank control group(p=0.041,24 h;0.038,48 h;0.03,72 h;0.011,96 h).The OD value(490nm)of thecells in the inhibitor control group at each time point were all not significantlydifferent from that of the blank control group(p=0.42,24 h;0.584,48 h;0.72,72h;0.064,96 h).7.The early apoptotic rate of cells in the miR-205 inhibitor group was(17.22 ±5.67)%,which was higher than(4.18 ± 0.96)% of the blank control group(p=0.033),and(3.74 ± 1.23)% of the inhibitor control group(p=0.01).Thedifferences were significant of statistics.There was no significant difference inearly apoptosis rate between the inhibitor control group and the blank controlgroup(p= 0.651).8.The relative number of invasive cells in the miR-205 inhibitor group was(58 ±11),which was lower than(120 ± 25,p=0.017)in the blank control group.and inthe inhibitor control group was(117 ± 22,p =0.014),the difference wassignificant of statistics.There was no significant difference between the relativenumber of invasive cells in the inhibitor control group and the blank controlgroup(p=0.088).?Conclusions? 1.The miR-205 was highly expressed in cervical cancer tissues and HeLa cell line.The miR-205 levels are correlated with FIGO stage,lymph node metastasis inpatients with cervical cancer.2.The down-regulated expression of miR-205 in cervical cancer He La cells caninhibit cell proliferation,promote cell apoptosis,resist cell invasion.