中国实用医刊
中國實用醫刊
중국실용의간
CENTRAL PLAINS MEDICAL JOURNAL
2012年
21期
1-4
,共4页
王万忠%种瑞峰%罗宜人%牟光远%陈志强
王萬忠%種瑞峰%囉宜人%牟光遠%陳誌彊
왕만충%충서봉%라의인%모광원%진지강
基因转染%p21waf1%细胞增殖%肺癌
基因轉染%p21waf1%細胞增殖%肺癌
기인전염%p21waf1%세포증식%폐암
Gene transfection%p21waf1%Cell proliferation%Lung carcinoma
目的 探讨外源性p21waf1基因转染对人肺癌细胞系A549细胞增殖的影响.方法 用脂质体介导的方法将pIRES-p21waf1真核表达载体转染人肺癌细胞系A549分为pIRES-p21waf1转染组、pIRES-neo空载体组及未转染组,应用实时荧光定量RT-PCR、Western blot分别检测各组p21waf1基因mRNA及p21蛋白表达情况.MTT法检测细胞增殖情况,流式细胞仪分析细胞周期变化,所有数据进行统计学分析.结果 p21waf1转染组p21waf1mRNA和p21蛋白高表达,差异有统计学意义(P＜0.01)；p21waf1转染组A549细胞生长速度明显低于空载体组和未转染组(P＜0.01)；流式细胞仪观察到p21waf1转染组细胞发生G1/S阻滞,G1期细胞比例显著高于空载体组和未转染组[(76.68±2.56)％ vs (50.27±1.24)％、(49.89±1.31)％,P＜0.01)],S期比例显著低于空载体组和未转染组[(12.48±1.08)％vs(39.59±1.19)％、(41.56±1.17)％,P＜0.01],并出现凋亡峰.结论 p21waf1基因转染可以抑制人肺癌细胞系A549细胞增殖并能诱导其发生细胞凋亡,可能为肺癌的基因治疗提供一条新的途径.
目的 探討外源性p21waf1基因轉染對人肺癌細胞繫A549細胞增殖的影響.方法 用脂質體介導的方法將pIRES-p21waf1真覈錶達載體轉染人肺癌細胞繫A549分為pIRES-p21waf1轉染組、pIRES-neo空載體組及未轉染組,應用實時熒光定量RT-PCR、Western blot分彆檢測各組p21waf1基因mRNA及p21蛋白錶達情況.MTT法檢測細胞增殖情況,流式細胞儀分析細胞週期變化,所有數據進行統計學分析.結果 p21waf1轉染組p21waf1mRNA和p21蛋白高錶達,差異有統計學意義(P＜0.01)；p21waf1轉染組A549細胞生長速度明顯低于空載體組和未轉染組(P＜0.01)；流式細胞儀觀察到p21waf1轉染組細胞髮生G1/S阻滯,G1期細胞比例顯著高于空載體組和未轉染組[(76.68±2.56)％ vs (50.27±1.24)％、(49.89±1.31)％,P＜0.01)],S期比例顯著低于空載體組和未轉染組[(12.48±1.08)％vs(39.59±1.19)％、(41.56±1.17)％,P＜0.01],併齣現凋亡峰.結論 p21waf1基因轉染可以抑製人肺癌細胞繫A549細胞增殖併能誘導其髮生細胞凋亡,可能為肺癌的基因治療提供一條新的途徑.
목적 탐토외원성p21waf1기인전염대인폐암세포계A549세포증식적영향.방법 용지질체개도적방법장pIRES-p21waf1진핵표체재체전염인폐암세포계A549분위pIRES-p21waf1전염조、pIRES-neo공재체조급미전염조,응용실시형광정량RT-PCR、Western blot분별검측각조p21waf1기인mRNA급p21단백표체정황.MTT법검측세포증식정황,류식세포의분석세포주기변화,소유수거진행통계학분석.결과 p21waf1전염조p21waf1mRNA화p21단백고표체,차이유통계학의의(P＜0.01)；p21waf1전염조A549세포생장속도명현저우공재체조화미전염조(P＜0.01)；류식세포의관찰도p21waf1전염조세포발생G1/S조체,G1기세포비례현저고우공재체조화미전염조[(76.68±2.56)％ vs (50.27±1.24)％、(49.89±1.31)％,P＜0.01)],S기비례현저저우공재체조화미전염조[(12.48±1.08)％vs(39.59±1.19)％、(41.56±1.17)％,P＜0.01],병출현조망봉.결론 p21waf1기인전염가이억제인폐암세포계A549세포증식병능유도기발생세포조망,가능위폐암적기인치료제공일조신적도경.
Objective To investigate the effect of transfected p21waf1 gene on the proliferation of human lung carcinoma cells.Methods p21waf1gene packaged with lipofectin was transfected into the cells of a human lung carcinoma cell line A549,which stably expressed p21waf1 gene.Thed p21waf1 mRNA and protein expression of A549-p21waf1,and cells were detected by Real Time PCR and Western blot methods,respectively.The cell growth was detected by MTT.The cell cycle pattern was assayed by flow cytometry.Results After transfection of pIRES-p21waf1 the p21waf1 mRNA and protein of A549-p21waf1 were highly expressed as compared with other cells.Cell growth was obviously inhibited and resulted in an accumulation of cells in G1,presenting that the proportion of the cells in G1 phase was obviously increased from (49.89 ± 1.31) ％ and (50.27 ± 1.24) ％ to (76.68 ± 2.56) ％,and that of the cells in S phases was obviously decreased from (39.59 ± 1.19)％ and (41.56 ± 1.17)％ to (12.48 ± 1.08) ％ respectively,as compared with control group.Conclusions p21waf1 gene can inhibit the growth and induce apoptosis of human lung carcinoma cells,which could be a new method of gene therapy for the lung carcinoma in future.
