中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2011年
8期
523-526
,共4页
何向民%娄毅%宋清斌%李建华
何嚮民%婁毅%宋清斌%李建華
하향민%루의%송청빈%리건화
RNA干扰%胃肿瘤%腺癌%基因,肿瘤抑制%基因表达%细胞凋亡
RNA榦擾%胃腫瘤%腺癌%基因,腫瘤抑製%基因錶達%細胞凋亡
RNA간우%위종류%선암%기인,종류억제%기인표체%세포조망
RNA interference%Stomach neoplasms%Adenocarcinoma%Genes,Tumor suppressor%Gene expression%Apoptosis
目的 探讨RNA干扰沉默生长抑制因子1(ING1)基因表达对胃腺癌AGS细胞凋亡的影响。方法人胃腺癌细胞株AGS经常规培养后分为空白对照组、阴性对照组[转染阴性对照小干扰RNA(siRNA)序列]、siRNA组（转染特异性ING1 siRNA序列）。应用免疫荧光、定量PCR和免疫印迹方法检测转染后不同时间的ING1基因表达沉默效果。应用流式细胞技术检测ING1基因表达沉默对AGS细胞凋亡的影响。结果ING1主要在AGS细胞胞质中表达。在荧光定量PCR技术检测中将空白对照组的ING1基因表达量设为1，则转染后24和40 h阴性对照组的ING1基因相对表达量分别为0.88±0.16和0.92±0.13，siRNA组ING1基因相对表达量分别为0.38±0.09和0.17±0.06。空白对照组和阴性对照组比较，P值分别=0.78和0.82。空白对照组和siRNA组比较，P值均=0.01。阴性对照组和siRNA组比较，P值分别=0.02和0.01。免疫印迹技术检测结果显示，转染后40 h AGS细胞中ING1蛋白表达水平明显下调。空白对照组AGS细胞凋亡率为11.06％±0.97％，阴性对照组、siRNA组转染40 h后AGS细胞凋亡率为11.82％±0.69％和 6.70％±0.41％。空白对照组与siRNA组比较P=0.024。空白对照组与阴性对照组比较P=0.76。阴性对照组与siRNA组比较P=0.019。结论ING1在人胃癌细胞凋亡过程中发挥重要作用，可能成为胃癌基因治疗的新靶点。
目的 探討RNA榦擾沉默生長抑製因子1(ING1)基因錶達對胃腺癌AGS細胞凋亡的影響。方法人胃腺癌細胞株AGS經常規培養後分為空白對照組、陰性對照組[轉染陰性對照小榦擾RNA(siRNA)序列]、siRNA組（轉染特異性ING1 siRNA序列）。應用免疫熒光、定量PCR和免疫印跡方法檢測轉染後不同時間的ING1基因錶達沉默效果。應用流式細胞技術檢測ING1基因錶達沉默對AGS細胞凋亡的影響。結果ING1主要在AGS細胞胞質中錶達。在熒光定量PCR技術檢測中將空白對照組的ING1基因錶達量設為1，則轉染後24和40 h陰性對照組的ING1基因相對錶達量分彆為0.88±0.16和0.92±0.13，siRNA組ING1基因相對錶達量分彆為0.38±0.09和0.17±0.06。空白對照組和陰性對照組比較，P值分彆=0.78和0.82。空白對照組和siRNA組比較，P值均=0.01。陰性對照組和siRNA組比較，P值分彆=0.02和0.01。免疫印跡技術檢測結果顯示，轉染後40 h AGS細胞中ING1蛋白錶達水平明顯下調。空白對照組AGS細胞凋亡率為11.06％±0.97％，陰性對照組、siRNA組轉染40 h後AGS細胞凋亡率為11.82％±0.69％和 6.70％±0.41％。空白對照組與siRNA組比較P=0.024。空白對照組與陰性對照組比較P=0.76。陰性對照組與siRNA組比較P=0.019。結論ING1在人胃癌細胞凋亡過程中髮揮重要作用，可能成為胃癌基因治療的新靶點。
목적 탐토RNA간우침묵생장억제인자1(ING1)기인표체대위선암AGS세포조망적영향。방법인위선암세포주AGS경상규배양후분위공백대조조、음성대조조[전염음성대조소간우RNA(siRNA)서렬]、siRNA조（전염특이성ING1 siRNA서렬）。응용면역형광、정량PCR화면역인적방법검측전염후불동시간적ING1기인표체침묵효과。응용류식세포기술검측ING1기인표체침묵대AGS세포조망적영향。결과ING1주요재AGS세포포질중표체。재형광정량PCR기술검측중장공백대조조적ING1기인표체량설위1，칙전염후24화40 h음성대조조적ING1기인상대표체량분별위0.88±0.16화0.92±0.13，siRNA조ING1기인상대표체량분별위0.38±0.09화0.17±0.06。공백대조조화음성대조조비교，P치분별=0.78화0.82。공백대조조화siRNA조비교，P치균=0.01。음성대조조화siRNA조비교，P치분별=0.02화0.01。면역인적기술검측결과현시，전염후40 h AGS세포중ING1단백표체수평명현하조。공백대조조AGS세포조망솔위11.06％±0.97％，음성대조조、siRNA조전염40 h후AGS세포조망솔위11.82％±0.69％화 6.70％±0.41％。공백대조조여siRNA조비교P=0.024。공백대조조여음성대조조비교P=0.76。음성대조조여siRNA조비교P=0.019。결론ING1재인위암세포조망과정중발휘중요작용，가능성위위암기인치료적신파점。
Objective To explore the effects of RNAi-mediated inhibitor of growth 1 (ING1)gene silencing on cell apoptosis of human gastric cancer cell line AGS.Methods After cultured,human gastric cell line AGS was divided into blank control group, negative control group (transfected with negative control siRNA sequence) and siRNA group (transfected with specific ING1 siRNA sequence).After transfection, the ING1 gene silencing effects at different time points were detected by immunofluorescence, real-time PCR and Western blot.The effects of ING1 gene silencing on apoptosis of AGS cells were evaluated by flow cytometry.Resnlts ING1 expression was restricted to the AGS cytoplasm.In real time PCR testing, the ING1 gene expression in blank control group was set as 1.24 h and 40 h after transfection, the ING1 gene relative expression quantity of negative control group was 0.88±0.16 and 0.92±0.13, and that of siRNA group was 0.38±0.09 and 0.17±0.06,respectively.Compared blank control group with negative control group, P values were 0.78 and 0.82.Compared blank control group with siRNA group, both P values were 0.01.Compared negative control group with siRNA group, P value was 0.02 and 0.01.In western blot testing, the protein expression of ING1 decreased significantly after transfected for 40 h.The apoptosis rate of blank control group was 11.06％ ±0.97％, and 40 h after transfection, the apoptosis rate in AGS cells of negative control group and siRNA group was 11.82 ％ ± 0.69 ％ and 6.70％ ± 0.41％, respectively.Compared negative control group with siRNA group, the P value was 0.024.Compared blank control group with negative control group, the P value was 0.76.Compared negative control group with siRNA group, the P value was 0.019.Conclusion ING1 plays an important role in cell apoptosis of human gastric adenocarcinoma cells, and may become a new target for gastric cancer gene therapy.
