中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
6期
1196-1198
,共3页
孟政雷%顾朝辉%田凤艳%贾占奎%李冠儒%孙科%曾甫清%杨锦建
孟政雷%顧朝輝%田鳳豔%賈佔奎%李冠儒%孫科%曾甫清%楊錦建
맹정뢰%고조휘%전봉염%가점규%리관유%손과%증보청%양금건
肾细胞癌%肾损伤分子-1%小干扰RNA%增殖%细胞周期
腎細胞癌%腎損傷分子-1%小榦擾RNA%增殖%細胞週期
신세포암%신손상분자-1%소간우RNA%증식%세포주기
Renal cell carcinoma%Kidney Injury molecular-1%Small interfering RNA%Proliferation%Cell cycle
目的 观察肾损伤分子-1(Kim-1)特异性小干扰RNA(siRNA)沉默Kim-1分子后,对786-0细胞增殖和细胞周期的影响.方法 利用脂质体转染方法将对人Kim-1基因序列特异的siRNA (50 nmol/L)转染进入786-0细胞中,转染后48 h采用实时定量聚合酶链反应(Real-time PCR)检测Kim-1基因的表达,采用噻唑蓝(MTT)比色法检测细胞增殖,转染后48 h采用流式细胞仪检测细胞周期.结果 Kim-1-siRNA能有效下调786-0细胞中Kim-1基因的表达水平,抑制细胞生长,使细胞周期阻滞在G0/G1期[3组转染后96 h的增殖抑制率分别为0、(3.80±1.24)％、(54.31±3.82)％,G0/G1期所占的比例分别为(38.44±1.82)％、(42.06±2.15)％、(63.69±2.87)％],差异有统计学意义(P＜0.05).结论 siRNA能有效抑制786-0细胞中Kim-1基因的表达,Kim-1基因表达下调影响786-0细胞的增殖并将细胞周期阻滞于G0/G1期.
目的 觀察腎損傷分子-1(Kim-1)特異性小榦擾RNA(siRNA)沉默Kim-1分子後,對786-0細胞增殖和細胞週期的影響.方法 利用脂質體轉染方法將對人Kim-1基因序列特異的siRNA (50 nmol/L)轉染進入786-0細胞中,轉染後48 h採用實時定量聚閤酶鏈反應(Real-time PCR)檢測Kim-1基因的錶達,採用噻唑藍(MTT)比色法檢測細胞增殖,轉染後48 h採用流式細胞儀檢測細胞週期.結果 Kim-1-siRNA能有效下調786-0細胞中Kim-1基因的錶達水平,抑製細胞生長,使細胞週期阻滯在G0/G1期[3組轉染後96 h的增殖抑製率分彆為0、(3.80±1.24)％、(54.31±3.82)％,G0/G1期所佔的比例分彆為(38.44±1.82)％、(42.06±2.15)％、(63.69±2.87)％],差異有統計學意義(P＜0.05).結論 siRNA能有效抑製786-0細胞中Kim-1基因的錶達,Kim-1基因錶達下調影響786-0細胞的增殖併將細胞週期阻滯于G0/G1期.
목적 관찰신손상분자-1(Kim-1)특이성소간우RNA(siRNA)침묵Kim-1분자후,대786-0세포증식화세포주기적영향.방법 이용지질체전염방법장대인Kim-1기인서렬특이적siRNA (50 nmol/L)전염진입786-0세포중,전염후48 h채용실시정량취합매련반응(Real-time PCR)검측Kim-1기인적표체,채용새서람(MTT)비색법검측세포증식,전염후48 h채용류식세포의검측세포주기.결과 Kim-1-siRNA능유효하조786-0세포중Kim-1기인적표체수평,억제세포생장,사세포주기조체재G0/G1기[3조전염후96 h적증식억제솔분별위0、(3.80±1.24)％、(54.31±3.82)％,G0/G1기소점적비례분별위(38.44±1.82)％、(42.06±2.15)％、(63.69±2.87)％],차이유통계학의의(P＜0.05).결론 siRNA능유효억제786-0세포중Kim-1기인적표체,Kim-1기인표체하조영향786-0세포적증식병장세포주기조체우G0/G1기.
Objective To investigate the effect of silencing kidney injury molecular-1 (Kim-1) gene by small interfering RNA (siRNA) interference on proliferation and cell cycle of renal cell carcinoma 786-0 cells.Methods The siRNA (50 nmol/L) which targets human Kim-1 gene was transfected into 786-0 cells by lipofectamine TM 2000.After 48 h,the expression of Kim-1 was detected by quantitative realtime quantitative polymerase chain reaction (Real-time PCR) quantitatively.The proliferation ability of cells in each group was determined by methyhhiazoltetrazolium (MTT) assay respectively.After 48 h,the cell cycle was examined by flow cytometry.Results In 786-0 cells the siRNA which targets Kim-1 gene silenced the expression of Kim-1 significantly,inhibited the proliferation and induced cell cycle arrest at G0/ G1 phases [for proliferation-inhibition rate 0,(3.80 ± 1.24) ％,(54.31 ± 3.82) ％ respectively ; rate in G0/G1 phases (38.44 ± 1.82) ％,(42.06 ± 2.15) ％,(63.69 ± 2.87) ％ respectively (P ＜ 0.05)].Conclusion In 786-0 cells,Kim-1 expression can be inhibited significantly by siRNA,and the down-regulation of Kim-1 expression could induce the proliferation inhibition and arrest of cell cycle at G0/G1 phases.
