miR-205对肾上腺皮质癌SW-13细胞转移潜能影响的研究  

Effects of miR-205 on proliferation,apoptosis and invasive abilities of adrenocortical carcinoma cell line SW-13

作　　者：吴义高[1] 王尉[1] 胡卫列[1] 肖戈[1] 徐文清[1] 

机构地区：[1]广州军区广州总医院泌尿外科.全军泌尿外科中心.广州军区泌尿外科研究所,广东广州510010

出　　处：《中华肿瘤防治杂志》2013年第8期566-571,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172421);国家自然科学基金青年科学基金(81001142)

摘　　要：目的:通过调控miR-205在肾上腺皮质癌(ACC)细胞株SW-13中的表达,探讨miR-205对SW-13细胞生物学行为的影响。方法:通过基因重组技术构建pcDNA3.1(+)-miR-205重组质粒并合成miR-205的反义寡核苷酸作为反向对照,调控miR-205在肾上腺皮质癌SW-13细胞中表达,实时荧光定量PCR法验证miR-205表达情况;分别采用MTS、EdU、TUNNEL及Transwell小室法,检测过表达及抑制miR-205对增殖、凋亡及侵袭的影响。结果:在转染pcDNA3.1(+)-miR-205及miR-205反义寡核苷酸后,SW-13细胞中miR-205分别上调了63.2倍(P=0.001)及下调了86.3%(P=0.002)。MTS结果显示,在24、48、72和96h时,miR-205上调后SW-13细胞增殖率下降了(10.50±1.80)%、(23.67±4.80)%、(30.9±5.40)%和(38.34±6.20)%,P值分别为0.041、0.014、0.016和0.032;miR-205下调后增殖率提高了(25.31±3.20)%、(32.51±4.60)%、(40.15±4.10)%和(38.34±6.20)%,P值分别为0.032、0.026、0.022和0.013。TUNNEL结果显示,pcDNA3.1(+)-miR-205组SW-13细胞凋亡数为(46±4)个,pcDNA3.1(+)组为(25±3)个,P=0.001 9;anti-miR-205转染组为(11±2)个,空白组为(25±3)个,P=0.002 5。Transwell侵袭实验显示,pcDNA3.1(+)-miR-205组穿过小室膜细胞数为(43±8)个,pcDNA3.1(+)组为(120±15)个,P=0.001 4;anti-miR-205转染组为(118±14)个,空白组为(220±20)个,P=0.001 9。结论:miR-205通过抑制细胞增殖、侵袭并促进凋亡在ACC SW-13细胞中起抑癌作用;成功构建pcD-NA3.1(+)-miR-205真核表达质粒,为进一步研究miR-205在SW-13细胞中的基因调控机制奠定基础。OBJECTIVE：To investigate the effect of miR-205 on SW-13 cell proliferation,apoptosis and invasive abilities by regulating the expression of miR-205 in adrenocortical carcinoma SW-13 cell line. METHODS： To regulate the expression of miR-205 in adrenocortical carcinoma SW-13 cell line by recombinant DNA technology to construct the pcD-NA3. 1（-4-）-miR-205 recombinant plasmid and synthesizing antisense oligonucleotides of miR-205 as the reverse control, The expression of miR-205 mRNA was detected by real-time fluorogenic quantitative-PCR（qRT-PCR）. The effects of pcDNA3.1 （＋）-miR-205 on SW-13 cell proliferation,apoptosis and invasive abilities were detected by MTS, EdU, TUN- NEL and Transwell chamber methods,respectively. RESULTS： qRT-PCR results revealed that miR-205 was increased by 63.2 fold （P=0. 001） and reduced by 86.3% （P=0. 002） in SW-13 cells after pcDNA3.1 （＋）-miR-205 and miR-205 antisense oligonucleotides transfected respectively. The MTS results suggested that the rate of cell proliferation of SW-13 was declined （10.50土1.80）%, （23.67土4.80）%, （30.79土5.40）% and （38. 34土6.20）% after the upreguhition of miR- 205 in 24,48,72 and 96 h （P values were 0. 041,0. 014,0. 016 and 0. 032）, while the rate was increased （25. 31土3.20） %, （32.51土4.60）%, （40.15土4.10）% and （38.34土6.20）% after the downregulation of miR-205 （P values were 0. 032,0. 026,0. 022 and 0. 013）. The TUNNEL results indicated that the number of SW-13 apoptotic cells were （46土4） and （25＋3）per vision in pcDNA3.1 （＋）-miR-205 group and pcDNA3.1 （＋） group （P=0. 001 9） ,While （11土2） and（25土3） per vision in anti-miR-205 group and blank group （P=0. 002 5）. Transwell invasion assay found that the number of SW-13 cells through the small room film were （43土8） and （120土15） per vision in pcDNA3.1 （＋）-miR-205 group and pcD- NA3.1 （＋） group （P=0. 001 4）, while （118土14） and （220土20） per vision in

关 键 词：肾上腺皮质肿瘤 生物学 MIRNA SW-13细胞 miR-205细胞凋亡 

分 类 号：R736.6[医药卫生—肿瘤]