miR-26a通过调节TFAP2C表达而抑制卵巢癌细胞的增殖  

miR-26a inhibits cell proliferation by regulating TFAP2C expression in ovarian cancer cells

作　　者：王淑芬[1] 董巍蕾 谢晶[1] 何璐[1] 周曦[1] 蔡艳林[1] 刘杰[1] 谢宛玉[1] 

机构地区：[1]南华大学附属第一医院妇产科,湖南衡阳421001

出　　处：《肿瘤》2014年第10期908-912,956,共6页Tumor

摘　　要：目的 :探讨微小RNA(microRNA,miRNA,miR)-26a通过调节转录因子活化蛋白2C(transcription factor activator protein 2C,TFAP2C)的表达而抑制卵巢癌SKOV3细胞的增殖。方法 :将miR-26a模拟物(miR-26a mimic)片段和含野生型TFAP2C基因3’-非翻译区的荧光素酶报告载体(TFAP2C-wt)共转染至SKOV3细胞后,进行荧光素酶活性检测。将miR-26a mimic、靶向TFAP2C基因的短发夹RNA(short hairpin RNA,shRNA)重组载体shRNA-TFAP2C和TFAP2C过表达的重组载体pcDNA3.1-TFAP2C分别转染或不同组合共转染至SKOV3细胞后,蛋白质印迹法检测细胞中TFAP2C的表达水平,MTS法检测细胞的增殖情况。结果 :miR-26a mimic和TFAP2C-wt共转染后,SKOV3细胞的荧光素酶活性强度比对照组[miRNA无关序列阴性对照(miRNA negative control,miR-NC)和含突变型TFAP2C基因3’-非翻译区的荧光素酶报告载体(TFAP2C-mut)共转染]下降了约52%(P<0.05)。miR-26a mimic转染组和shRNA-TFAP2C转染组SKOV3细胞中TFAP2C蛋白的表达水平和细胞增殖率均低于相应的对照组(miR-NC和shRNA-NC转染SKOV3细胞),差异有统计学意义(P<0.05);而miR-26a mimic和shRNA-TFAP2C共转染组SKOV3细胞的增殖率低于miR-26a mimic和shRNA-TFAP2C单独转染组,差异有统计学意义(P<0.05)。pcDNA3.1-TFAP2C转染组SKOV3细胞中TFAP2C的表达水平和细胞增殖率高于相应的对照组(pcDNA3.1空载体转染SKOV3细胞),差异有统计学意义(P<0.05);而miR-26a mimic和pcDNA3.1-TFAP2C共转染组SKOV3细胞的增殖率低于pcDNA3.1-TFAP2C转染组,差异有统计学意义(P<0.05)。结论 :miR-26a通过调控TFAP2C的表达而抑制卵巢癌SKOV3细胞的增殖。Objective： To investigate the suppression effect of microRNA-26a（miR-26a） on the proliferation of ovarian cancer SKOV3 cells by regulating the expression of transcription factor activator protein 2C（TFAP2C）. Methods： The miR-26 a mimic and luciferase reporter vector TFAP2C-wt containing 3’-untranslated region（3’-UTR） of wild type TFAP2 C were co-transfected into SKOV3 cells, and the luciferase activity was detected. After the SKOV3 cells were transfected with miR-26 a mimic, short hairpin RNA（shRNA） targeting TFAP2 C gene recombination vector shRNA-TFAP2 C and TFAP2 C over-expression recombination vector pcDNA3.1-TFAP2 C, repectively, as well as cotransfected with the different combinations, the expression levels of TFAP2 C were detected by Western blotting, and the cell proliferation was examined by MTS assay. Results： As compared with the control [miRNA negative control（miR-NC） and luciferase reporter vector TFAP2C-mut containing 3’-UTR of mutant type of TFAP2 C gene were in co-transfection], the luciferase activity of SKOV3 cells after co-transfection with miR-26 a mimic and TFAP2C-wt was decreased by 52%（P 〈 0.05）. The expression levels of TFAP2 C and the proliferation rates of SKOV3 cells after transfection with miR-26 a mimic and shRNA-TFAP2 C were lower than those of the SKOV3 cells transfected with miR-NC and shRNA-NC（P 〈 0.05）. The proliferation rate of SKOV3 cells after co-transfection with miR-26 a mimic and shRNA-TFAP2 C was lower than those of the SKOV3 cells transfected with miR-26 a mimic and shRNA-TFAP2 C alone（P 〈 0.05）.The expression levels of TFAP2 C and the proliferation rate of SKOV3 cells after transfection with pcDNA3.1-TFAP2 C were higher than those of the SKOV3 cells transfected with empty vector pcDNA3.1（P 〈 0.05）. The proliferation rate of SKOV3 cells after co-transfection with miR-26 a mimic and pcDNA3.1-TFAP2 C was lower than that of the SKOV3 cells transfected with pcDNA3.1-TFAP2 C alone（P 〈 0.05）. Conclusion： miR-26 a c

关 键 词：卵巢肿瘤 微RNAS 细胞增殖 SKOV3细胞 

分 类 号：R737.31[医药卫生—肿瘤]