中国临床医学
中國臨床醫學
중국림상의학
CLINICAL MEDICAL JOURNAL OF CHINA
2014年
6期
643-646
,共4页
张素青%陈橼%张一心%刘继斌%朱烨菁%钱俊%李德春
張素青%陳櫞%張一心%劉繼斌%硃燁菁%錢俊%李德春
장소청%진연%장일심%류계빈%주엽정%전준%리덕춘
小干扰 RNA%YAP%肝癌细胞%阿霉素
小榦擾 RNA%YAP%肝癌細胞%阿黴素
소간우 RNA%YAP%간암세포%아매소
Small interfering RNA%Yes-associated protein%Hepatoma cells%Adriamycin
目的：研究用小片段干扰 RNA（small interfering RNA，siRNA）抑制肝癌细胞株中 YAP（yes-associated protein）表达后肝癌细胞株对阿霉素（adriamycin，ADM）的敏感性变化。方法：设计 YAP 基因的 siRNA 序列，用脂质体 LipofectamineTM 2000将 siRNA 转染至4株肝癌细胞株 PLC／PRF／5、Huh-7、MHCC 97H、MHCC 97L 中，在荧光显微镜下观察并采用蛋白免疫印迹（Western blotting）法检测肝癌细胞株中 YAP 的表达。将细胞以不同浓度的 ADM 处理后，采用四甲基偶氮唑蓝（methyl thialzolyl tetrazolium，MTT）比色法检测细胞的存活率。结果：成功构建针对 YAP 的 siRNA 表达载体。Western blotting 结果显示，肝癌细胞株经特异性 siRNA 转染后，YAP 蛋白的表达受到抑制，表明 RNA 干扰（RNA interference， RNAi）能特异、有效地抑制肝癌细胞株中 YAP 的表达。MTT 结果显示，转染 YAP siRNA 后肝癌细胞株的存活率受到明显抑制。结论：siRNA 抑制肝癌细胞株中 YAP 的表达可增强肝癌细胞株对 ADM 的敏感性。
目的：研究用小片段榦擾 RNA（small interfering RNA，siRNA）抑製肝癌細胞株中 YAP（yes-associated protein）錶達後肝癌細胞株對阿黴素（adriamycin，ADM）的敏感性變化。方法：設計 YAP 基因的 siRNA 序列，用脂質體 LipofectamineTM 2000將 siRNA 轉染至4株肝癌細胞株 PLC／PRF／5、Huh-7、MHCC 97H、MHCC 97L 中，在熒光顯微鏡下觀察併採用蛋白免疫印跡（Western blotting）法檢測肝癌細胞株中 YAP 的錶達。將細胞以不同濃度的 ADM 處理後，採用四甲基偶氮唑藍（methyl thialzolyl tetrazolium，MTT）比色法檢測細胞的存活率。結果：成功構建針對 YAP 的 siRNA 錶達載體。Western blotting 結果顯示，肝癌細胞株經特異性 siRNA 轉染後，YAP 蛋白的錶達受到抑製，錶明 RNA 榦擾（RNA interference， RNAi）能特異、有效地抑製肝癌細胞株中 YAP 的錶達。MTT 結果顯示，轉染 YAP siRNA 後肝癌細胞株的存活率受到明顯抑製。結論：siRNA 抑製肝癌細胞株中 YAP 的錶達可增彊肝癌細胞株對 ADM 的敏感性。
목적：연구용소편단간우 RNA（small interfering RNA，siRNA）억제간암세포주중 YAP（yes-associated protein）표체후간암세포주대아매소（adriamycin，ADM）적민감성변화。방법：설계 YAP 기인적 siRNA 서렬，용지질체 LipofectamineTM 2000장 siRNA 전염지4주간암세포주 PLC／PRF／5、Huh-7、MHCC 97H、MHCC 97L 중，재형광현미경하관찰병채용단백면역인적（Western blotting）법검측간암세포주중 YAP 적표체。장세포이불동농도적 ADM 처리후，채용사갑기우담서람（methyl thialzolyl tetrazolium，MTT）비색법검측세포적존활솔。결과：성공구건침대 YAP 적 siRNA 표체재체。Western blotting 결과현시，간암세포주경특이성 siRNA 전염후，YAP 단백적표체수도억제，표명 RNA 간우（RNA interference， RNAi）능특이、유효지억제간암세포주중 YAP 적표체。MTT 결과현시，전염 YAP siRNA 후간암세포주적존활솔수도명현억제。결론：siRNA 억제간암세포주중 YAP 적표체가증강간암세포주대 ADM 적민감성。
Objective:To investigate the inhibitory effect of small interfering RNA(siRNA)on yes-associated protein(YAP) expression and adriamycin(ADM)chemosensitivity of hepatoma cell lines.Methods:The siRNA sequences were designed based on YAP gene and were transfected into four different kinds of hepatoma cells (PLC/PRF/5,Huh-7,MHCC 97H,MHCC 97L) through lipofectamineTM 2000.The inhibition result of YAP expression in hepatoma cells was measured by Western blotting and fluorescence microscope.The viability rate of hepatoma cells was measured by methyl thiazolyl tetrazolium(MTT)colori-metry after treating with different concentrations of ADM.Results:The siRNA expression vector to aim directly at YAP gene was constructed successfully.Western blotting revealed that transfection with siRNA could inhibit the expression of YAP pro-tein in hepatoma cell lines.The expression of YAP protein in hepatoma cell lines was specially and effectually inhibited by RNA interference(RNAi).MTT colorimetry detection result revealed that the viability rate of hepatoma cell lines significantly de-creased after transfection with YAP siRNA.Conclusions:Inhibiting the expression of YAP by siRNA can enhance the sensitivi-ty of hepatoma cell lines to ADM.
