中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2013年
9期
1195-1198
,共4页
张君薇%张毅%杜贾军%刘奇
張君薇%張毅%杜賈軍%劉奇
장군미%장의%두가군%류기
RNA干扰%DNA结合蛋白质类/遗传学%反式激活因子类/遗传学%肿瘤细胞,培养的%肝肿瘤/病理学%细胞凋亡
RNA榦擾%DNA結閤蛋白質類/遺傳學%反式激活因子類/遺傳學%腫瘤細胞,培養的%肝腫瘤/病理學%細胞凋亡
RNA간우%DNA결합단백질류/유전학%반식격활인자류/유전학%종류세포,배양적%간종류/병이학%세포조망
RNA interference%DNA-binding proteins/genetics%Trans-activators/genetics%Tumor cells,cultured%Liver neoplasms/pathology%Apoptosis
目的 探讨JAK/STAT3信号转导途径对肝癌细胞凋亡的影响.方法 以BEL-7402肝癌细胞为模型,采用RNAi技术,将STAT3-siRNA转染BEL-7402肝癌细胞,沉默STAT3基因表达,另设细胞对照组,阴性质粒组(pGCsi.U6/neoRFP scramble-siRNA载体转染BEL-7402).分别通过流式细胞术检测细胞凋亡比率的变化,JC-1荧光染色观察线粒体膜电势Δψm变化,并利用Western Blot方法检测Caspase 3蛋白质水平的变化.结果 STAT3-siRNA组凋亡率为(38.82 ±0.88)％,明显高于其他组[对照组(9.22±0.38)％,阴性质粒组(16.47±1.04)％,P＜0.05],并引起细胞线粒体膜电势明显降低[(59.06±1.89)％vs(91.33±1.78)％和(89.90±1.92)％,P＜0.05],活性Caspase 3蛋白在STAT3-siRNA组表达明显高于其他组(0.48±0.05 vs 0.22±0.04和0.26 ±0.06,P＜ 0.05).结论 RNAi沉默STAT3基因,抑制JAK/STAT3信号转导途径,促进BEL-7402肝癌细胞凋亡.
目的 探討JAK/STAT3信號轉導途徑對肝癌細胞凋亡的影響.方法 以BEL-7402肝癌細胞為模型,採用RNAi技術,將STAT3-siRNA轉染BEL-7402肝癌細胞,沉默STAT3基因錶達,另設細胞對照組,陰性質粒組(pGCsi.U6/neoRFP scramble-siRNA載體轉染BEL-7402).分彆通過流式細胞術檢測細胞凋亡比率的變化,JC-1熒光染色觀察線粒體膜電勢Δψm變化,併利用Western Blot方法檢測Caspase 3蛋白質水平的變化.結果 STAT3-siRNA組凋亡率為(38.82 ±0.88)％,明顯高于其他組[對照組(9.22±0.38)％,陰性質粒組(16.47±1.04)％,P＜0.05],併引起細胞線粒體膜電勢明顯降低[(59.06±1.89)％vs(91.33±1.78)％和(89.90±1.92)％,P＜0.05],活性Caspase 3蛋白在STAT3-siRNA組錶達明顯高于其他組(0.48±0.05 vs 0.22±0.04和0.26 ±0.06,P＜ 0.05).結論 RNAi沉默STAT3基因,抑製JAK/STAT3信號轉導途徑,促進BEL-7402肝癌細胞凋亡.
목적 탐토JAK/STAT3신호전도도경대간암세포조망적영향.방법 이BEL-7402간암세포위모형,채용RNAi기술,장STAT3-siRNA전염BEL-7402간암세포,침묵STAT3기인표체,령설세포대조조,음성질립조(pGCsi.U6/neoRFP scramble-siRNA재체전염BEL-7402).분별통과류식세포술검측세포조망비솔적변화,JC-1형광염색관찰선립체막전세Δψm변화,병이용Western Blot방법검측Caspase 3단백질수평적변화.결과 STAT3-siRNA조조망솔위(38.82 ±0.88)％,명현고우기타조[대조조(9.22±0.38)％,음성질립조(16.47±1.04)％,P＜0.05],병인기세포선립체막전세명현강저[(59.06±1.89)％vs(91.33±1.78)％화(89.90±1.92)％,P＜0.05],활성Caspase 3단백재STAT3-siRNA조표체명현고우기타조(0.48±0.05 vs 0.22±0.04화0.26 ±0.06,P＜ 0.05).결론 RNAi침묵STAT3기인,억제JAK/STAT3신호전도도경,촉진BEL-7402간암세포조망.
Objective To explore influence of JAK/STAT3 signaling pathway on the apoptosis of HCC cells.Methods DNA-vector-based RNAi approach silence was used to down-regulate STAT3 expression in Bel-7402 cells.According to the STAT3 cDNA sequence in the GeneBank database,the plasmid pGCsi.U6/neoRFP-STAT3 that was designed for expression of STAT3 siRNA was constructed and synthesized,and then transfected into the Bel-7402 cells with iipofectamine 2000.The apoptotic rate was measured with flow cytometry (FCM) and annexinV/PI apoptosis detection kit staining.The mitochondrial membrane potential (BΨm) was visualized by the JC-1 fluorescence staining and the inverted fluorescence microscope.Moreover,the expression of caspase-3 protein was analyzed by Western blotting.Results The apoptotic ratio of STAT3-siRNA group was (38.82 ± 0.88) ％,which was significantly higher than that in other group [control group (9.22 ± 0.38) ％,scramble-siRNA group (16.47 ± 1.04) ％,P ＜ 0.05].The mitochondrial membrane potential of STAT3-siRNA group observed by the JC-1 fluorescence staining was decreased significantly[(91.33 ± 1.78) ％] and [(89.90 ± 1.92) ％ vs (59.06 ± 1.89) ％,P ＜ 0.05].The Western blot results showed that the protein expression of active caspase-3 in STAT3-siRNA group was significantly higher than other groups (0.48 ± 0.05 vs 0.22 ± 0.04 and 0.26 ± 0.06,P ＜ 0.05).Conclusions STAT3 gene silencing significantly improves the apoptotic effect in the Bel-7402 ceils.
