郑州大学学报（医学版）
鄭州大學學報（醫學版）
정주대학학보（의학판）
JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES)
2015年
5期
641-644
,共4页
阙菡雅%王小龙%李杰%徐小艳%夏艳秋%周舫
闕菡雅%王小龍%李傑%徐小豔%夏豔鞦%週舫
궐함아%왕소룡%리걸%서소염%하염추%주방
肺癌%A549细胞%siRNA%热休克蛋白70%JNK%凋亡
肺癌%A549細胞%siRNA%熱休剋蛋白70%JNK%凋亡
폐암%A549세포%siRNA%열휴극단백70%JNK%조망
lung cancer%A549 cell%siRNA%heat shock protein 70%JNK%apoptosis
目的：探讨沉默热休克蛋白70（ HSP70）的表达对人肺腺癌A549细胞凋亡的影响。方法：将体外培养的A549细胞分为空白对照组（仅加入转染试剂Lipofectamine 2000）、阴性对照组（转染无关序列）和干扰组（转染HSP70 siRNA）3组，采用流式细胞仪检测A549细胞凋亡率的变化，采用实时定量PCR法检测各组细胞中HSP70 mRNA表达的差异，采用Western blot 检测各组细胞中HSP70、p-JNK及Caspase-3蛋白表达的变化。结果：转染后24 h，干扰组A549细胞中HSP70 mRNA及蛋白表达明显受到抑制，与空白对照组和阴性对照组相比差异有统计学意义（F＝150．348和450．953，P均＜0．001）；转染48 h后，干扰组细胞凋亡率高于空白对照组和阴性对照组，差异有统计学意义（F＝229．636，P＜0．001）。与空白对照组和阴性对照组相比，干扰组A549细胞中p-JNK蛋白和Caspase-3蛋白表达明显受到抑制，差异有统计学意义（F＝877．094和340．808，P均＜0．001）。结论：沉默HSP70的表达可促进A549细胞的凋亡，且该过程伴随着p-JNK和Caspase-3表达水平的降低。
目的：探討沉默熱休剋蛋白70（ HSP70）的錶達對人肺腺癌A549細胞凋亡的影響。方法：將體外培養的A549細胞分為空白對照組（僅加入轉染試劑Lipofectamine 2000）、陰性對照組（轉染無關序列）和榦擾組（轉染HSP70 siRNA）3組，採用流式細胞儀檢測A549細胞凋亡率的變化，採用實時定量PCR法檢測各組細胞中HSP70 mRNA錶達的差異，採用Western blot 檢測各組細胞中HSP70、p-JNK及Caspase-3蛋白錶達的變化。結果：轉染後24 h，榦擾組A549細胞中HSP70 mRNA及蛋白錶達明顯受到抑製，與空白對照組和陰性對照組相比差異有統計學意義（F＝150．348和450．953，P均＜0．001）；轉染48 h後，榦擾組細胞凋亡率高于空白對照組和陰性對照組，差異有統計學意義（F＝229．636，P＜0．001）。與空白對照組和陰性對照組相比，榦擾組A549細胞中p-JNK蛋白和Caspase-3蛋白錶達明顯受到抑製，差異有統計學意義（F＝877．094和340．808，P均＜0．001）。結論：沉默HSP70的錶達可促進A549細胞的凋亡，且該過程伴隨著p-JNK和Caspase-3錶達水平的降低。
목적：탐토침묵열휴극단백70（ HSP70）적표체대인폐선암A549세포조망적영향。방법：장체외배양적A549세포분위공백대조조（부가입전염시제Lipofectamine 2000）、음성대조조（전염무관서렬）화간우조（전염HSP70 siRNA）3조，채용류식세포의검측A549세포조망솔적변화，채용실시정량PCR법검측각조세포중HSP70 mRNA표체적차이，채용Western blot 검측각조세포중HSP70、p-JNK급Caspase-3단백표체적변화。결과：전염후24 h，간우조A549세포중HSP70 mRNA급단백표체명현수도억제，여공백대조조화음성대조조상비차이유통계학의의（F＝150．348화450．953，P균＜0．001）；전염48 h후，간우조세포조망솔고우공백대조조화음성대조조，차이유통계학의의（F＝229．636，P＜0．001）。여공백대조조화음성대조조상비，간우조A549세포중p-JNK단백화Caspase-3단백표체명현수도억제，차이유통계학의의（F＝877．094화340．808，P균＜0．001）。결론：침묵HSP70적표체가촉진A549세포적조망，차해과정반수착p-JNK화Caspase-3표체수평적강저。
Aim:To explore the effect of heat shock protein 70(HSP70) by small interfering RNA (siRNA) on the apoptosis in human lung cancer A549 cells.Methods: A549 cells were allocated into 3 groups,among them,the mock group was treated with Lipofectamine 2000 only, the negative control(NC) group was transfected with NC-siRNA and the HSP70 siRNA group was transfected with HSP 70 siRNA.The cell apoptosis rate was tested by flow cytometry .HSP70 mR-NA level was tested by real-time PCR.The expressions of HSP70, p-JNK and Caspase-3 protein were detected by Western blot.Results:After being transfected for 24 h, the HSP70 mRNA and protein levels in HSP70 siRNA group were de-creased compared with mock group and NC group (F=150.348, 450.953,P<0.001).After being transfected for 48 h, the apoptosis rate in HSP70 siRNA group was increased compared with mock group and NC group (F =229.636,P <0.001),the levels of p-JNK and Caspase-3 protein were decreased compared with mock group and NC group (F=877.094, 340.808,P<0.001).Conclusion: Suppression of HSP70 by siRNA can promote apoptosis of A549 cells accompanying the decrease of the activation of p-JNK and Caspase-3.
