Expression of microRNA 27a and its correlation with drug resistance in human ovarian cancer A2780/Taxol cells
10.3760/cma.j.issn.0529-567x.2010.05.012
- VernacularTitle:微小RNA 27a在卵巢上皮性癌紫杉醇耐药细胞中的表达及其与耐药的关系
- Author:
Zhimin LI
;
Sha HU
;
Lan XIAO
;
Jing WANG
;
Jing CAI
;
Lili YU
;
Zehua WANG
- Publication Type:Journal Article
- Keywords:
Ovarian neoplasms;
Paclitaxel;
MicroRNAs;
Drug resistance,neoplasm
- From:
Chinese Journal of Obstetrics and Gynecology
2010;45(5):372-375
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells.Methods A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells.The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA ( NC) by lipofectamine 2000.The expressions of MDR1 gene,P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively.Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity.Apoptosis analysis was measured by fluorescence activated cell sorter ( FACS).Results (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells,with a significant difference between the two groups (P <0.05).(2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%,P-gp protein level[(26 ±5)%]decreased than that in the NC group[(43 ±7)%],HIPK2 protein level[(30 ±6)%]increased than that in the NC group[(19 ±4)%],the 50% inhibitionconcentration (0.5 (μmol/L) was less than that in the NC group (6.8 μmol/L),apoptosis rate[(32.5 ± 3.6) %]was higher than that in the NC group[(5.6 ±2.1) %],and there were significant differences between two groups (all P < 0.05 ).( 3 ) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05).Conclusion The expression of miR-27a is upregulated in A2780/Taxol cells,which may regulate MDR1 and P-gp expression by targeting HIPK2.
