中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
9期
2169-2171
,共3页
全红%白俊超%高玮%吕莉%赵中辛%韩俊毅
全紅%白俊超%高瑋%呂莉%趙中辛%韓俊毅
전홍%백준초%고위%려리%조중신%한준의
含菱形结构蛋白1%结肠癌%RNA干扰%增殖
含蔆形結構蛋白1%結腸癌%RNA榦擾%增殖
함릉형결구단백1%결장암%RNA간우%증식
Rhomboid domain containing 1%Colorectal carcinoma%RNA interference%Proliferation
目的 探讨含菱形结构蛋白1(RHBDD1)基因对结肠癌细胞增殖影响及机制.方法 使用慢病毒介导RNA干扰技术下调RKO细胞RHBDD1基因水平;噻唑蓝(MTT)法、克隆形成实验检测RKO细胞增殖变化;流式细胞仪检测细胞周期变化.结果 实时定量聚合酶链反应(Real-time PCR)检测Lv-shRHBDD1组荧光强度(0.49±0.35)明显降低,证实RNA干扰下调了RHBDD1基因水平(P＜0.01);MTT法显示RHBDD1基因下调后RKO细胞增殖率显著低于未下调细胞(P＜0.01);克隆形成试验显示Lv-shRHBDD1组RKO细胞克隆数(35.0±2.9)明显减少(P＜0.01);流式细胞仪分析显示Lv-shRHBDD1组G0/G1期[(50.20±1.22)％]细胞比例增多,S期[(39.30±0.87)％]和G/M期[(8.10±0.72)％]细胞比例下降,表明慢病毒介导下调RHBDD1基因的RKO细胞周期阻滞于G0/G1期(P＜0.01).结论 RHBDD1在结肠癌细胞增殖过程中发挥重要作用.
目的 探討含蔆形結構蛋白1(RHBDD1)基因對結腸癌細胞增殖影響及機製.方法 使用慢病毒介導RNA榦擾技術下調RKO細胞RHBDD1基因水平;噻唑藍(MTT)法、剋隆形成實驗檢測RKO細胞增殖變化;流式細胞儀檢測細胞週期變化.結果 實時定量聚閤酶鏈反應(Real-time PCR)檢測Lv-shRHBDD1組熒光彊度(0.49±0.35)明顯降低,證實RNA榦擾下調瞭RHBDD1基因水平(P＜0.01);MTT法顯示RHBDD1基因下調後RKO細胞增殖率顯著低于未下調細胞(P＜0.01);剋隆形成試驗顯示Lv-shRHBDD1組RKO細胞剋隆數(35.0±2.9)明顯減少(P＜0.01);流式細胞儀分析顯示Lv-shRHBDD1組G0/G1期[(50.20±1.22)％]細胞比例增多,S期[(39.30±0.87)％]和G/M期[(8.10±0.72)％]細胞比例下降,錶明慢病毒介導下調RHBDD1基因的RKO細胞週期阻滯于G0/G1期(P＜0.01).結論 RHBDD1在結腸癌細胞增殖過程中髮揮重要作用.
목적 탐토함릉형결구단백1(RHBDD1)기인대결장암세포증식영향급궤제.방법 사용만병독개도RNA간우기술하조RKO세포RHBDD1기인수평;새서람(MTT)법、극륭형성실험검측RKO세포증식변화;류식세포의검측세포주기변화.결과 실시정량취합매련반응(Real-time PCR)검측Lv-shRHBDD1조형광강도(0.49±0.35)명현강저,증실RNA간우하조료RHBDD1기인수평(P＜0.01);MTT법현시RHBDD1기인하조후RKO세포증식솔현저저우미하조세포(P＜0.01);극륭형성시험현시Lv-shRHBDD1조RKO세포극륭수(35.0±2.9)명현감소(P＜0.01);류식세포의분석현시Lv-shRHBDD1조G0/G1기[(50.20±1.22)％]세포비례증다,S기[(39.30±0.87)％]화G/M기[(8.10±0.72)％]세포비례하강,표명만병독개도하조RHBDD1기인적RKO세포주기조체우G0/G1기(P＜0.01).결론 RHBDD1재결장암세포증식과정중발휘중요작용.
Objective To investigate the role of rhomboid domain containing 1 (RHBDD1) in colorectal carcinoma (CRC) tumorigenesis.Methods Lentivirus-mediated RNA interference (RNAi) was employed to knock down RHBDD1 in RKO cells.RKO cell proliferation alteration was examined by methyl thiazol tetrazolium (MTT) and colony formation capacity with RKO monolayer cell culture was assessed;Cell cycle analysis was performed by flow cytometry.Results Real-time quantitative polymerase chain reaction (Real-time PCR) showed arbltrary unit of Lv-shRHBDD1 group decreased (0.49 ± 0.35),which confirmed knockdown efficiency (P ＜0.01).Functional analysis indicated that depletion of RHBDD1 could significantly down-regulate CRC cell proliferation (P ＜ 0.01).Colony formation capacity of Lv-shRHBDD1 group (35.0 ± 2.9) was significantly reduced (P ＜ 0.01).Flow cytometry showed that RKO cell proportion of Lv-shRHBDD1 group was increased in G0/G1 phase (35.0 ± 2.9),and decreased in S phase [(39.30 ± 0.87) ％] and G2/M phase [(8.10 ± 0.72) ％] (P ＜ 0.01).Conclusion It could be concluded from this study that RHBDD1 might contribute to CRC tumorigenesis and serve as a potential therapeutic target in human colon cancer.
