小分子干扰RNA沉默ACS5基因对结肠癌细胞表达及增殖的影响  

Effects of acyi-CoA synthetase 5 silencing by siRNA on expression and proliferation of colon carcinoma cell lines

作　　者：丁世华[1] 唐文芳[1] 帅红梅[1] 吴东海[1] 郭海建[1] 刘俊[1] 

机构地区：[1]深圳大学第一附属医院消化内科,深圳518035

出　　处：《中国医师杂志》2016年第12期1808-1811,共4页Journal of Chinese Physician

基　　金：深圳市科技创新委员会基础研究项目（JCYJ20140414170821217）

摘　　要：目的探讨小分子干扰RNA（siRNA）沉默脂酰辅酶A合成酶5（ACS5）基因对结肠癌细胞表达及增殖的影响。方法采用免疫组化检测30例结肠癌及癌旁组织ACS5的表达。设计合成siRNA．ACS5，用Lipofectamine2000TM搭载转染人结肠癌细胞HT-29、SW480，实时定量PCR检测ACS5mRNA表达量，四唑氮化合物（MTS）法检测转染后结肠癌细胞增殖情况。结果免疫组化显示结肠癌组织ACS5阳性率83．3％（25／30）明显高于癌旁组织30％（9／30）。结肠癌细胞HT-29siRNA—ACS5干扰组的mRNA的表达量（0．18±0．03）显著低于空白对照组（2．48±0．12）和NCsiRNA阴性对照组（2．55±0．31），抑制率92．96％（F=146．9，P〈0．01）。结肠癌细胞SW480siRNA—ACS5干扰组的mRNA的表达量（0．14±0．01）显著低于空白对照组（1±0．03）和NcsiRNA阴性对照组（1．21±0．05），抑制率88．5％（F=826．59，P〈0．01）。siRNA—ACs5干扰组在48、72h的增殖能力明显低于两个对照组（P〈0．05）；而两个对照组在各时间点比较差异均无统计学意义（P〉0．05）。结论ACS在结肠癌组织中高表达，沉默ACS基因可下调结肠癌细胞ACS表达并抑制其增殖。Objective To investigate the effects of acyl-CoA synthetase 5 （ACS5） silencing by siRNA on expression and proliferation of colon carcinoma cell lines. Methods The expression of ACS5 in 30 case colon carcinoma and adjacent tissues were analyzed by immunohistochemical staining. The siRNA of ACS5 with Lipofectamine2000TM was transfected into colon carcinoma cell lines （HT-29 and SW480）. The expression of ACS5 in colon carcinoma cell lines （HT-29 and SW480） was detected by real-time reverse transcription polymerase chain reaction. Proliferation of colon carcinoma cell lines was analyzed by 3-（4,5- dimenthylthiazol-2-yl） -5-（3-carboxymethoxy - phenyl） -2-（ 4-sulfophenyl ） -2H-tetrazolium （ MTS ）. Re- suits The expression of ACS5 in colon cancer was significantly higher than in adjacent tissues by immuno- histochemical staining. The mRNA of ACS5 in siRNA-ACS5 group （0. 18 ±0. 03 ） was significantly lower than in NC siRNA group （2.55 ±0.31 ） and blank control group （2.48 ± 0. 12） in HT-29 colon cancer lines, and the inhibition ratio was 92. 96% （F = 146.9,P 〈0. 01 ）. The mRNA of ACS5 in siRNA-ACS5 group （0. 14 ±0. 01 ） was significantly lower than in NC siRNA group （ 1.21 ± 0. 05 ） and blank control group （1 ±0. 03） in SW480 colon cancer lines, and the inhibition ratio was 88.5% （F = 826. 5.9, P 〈 0. 01 ）. Proliferation of HT-29 and SW480 colon cancer line in siRNA-ACS5 group was slower on 72 h and 96 h than in NC siRNA group and blank control group （P 〈 0. 05 ）. Conclusions Expression of ACS5 is elevated in colon cancer tissues, siRNA interference of colon cancer line downregulated ACS5 expression and inhibited the proliferation of the colon cancer cells.

关 键 词：RNA 小分子干扰 辅酶A连接酶类/代谢 结肠肿瘤/代谢/病理学 

分 类 号：R735.35[医药卫生—肿瘤]