中国医学创新
中國醫學創新
중국의학창신
MEDICAL INNOVATION OF CHINA
2013年
1期
3-5
,共3页
基因转染%p21waf1%细胞增殖%胃癌
基因轉染%p21waf1%細胞增殖%胃癌
기인전염%p21waf1%세포증식%위암
Gene transfection%p21waf1%Cell proliferation%Gastric cancer
目的:探讨外源性 p21waf1基因转染对人胃癌细胞系 BGC-823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胃癌细胞系 BGC-823,分为 pIRES-p21waf1转染组、pIRES-neo 空载体组及未转染组三个组别.应用实时荧光定量 RT-PCR、Western blot 免疫印迹分别在基因和蛋白两个水平检测各组 p21的表达情况,采用四甲基偶氮唑蓝(MTT)法测定细胞增殖能力,流式细胞仪分析细胞 DNA 周期变化,所有数据进行统计学分析.结果:p21waf1转染组 p21waf1 mRNA 和 p21蛋白高表达(P<0.01);p21waf1转染组BGC-823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转染组细胞的 G1期细胞比例显著高于空载体组和未转染组(P<0.01),S 期比例显著低于空载体组和未转染组(P<0.01),并出现凋亡峰.结论:p21waf1基因转染可以抑制人胃癌细胞系BGC-823细胞增殖并能诱导其发生细胞凋亡,可能为胃癌的基因治疗提供一条新的途径.
目的:探討外源性 p21waf1基因轉染對人胃癌細胞繫 BGC-823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胃癌細胞繫 BGC-823,分為 pIRES-p21waf1轉染組、pIRES-neo 空載體組及未轉染組三箇組彆.應用實時熒光定量 RT-PCR、Western blot 免疫印跡分彆在基因和蛋白兩箇水平檢測各組 p21的錶達情況,採用四甲基偶氮唑藍(MTT)法測定細胞增殖能力,流式細胞儀分析細胞 DNA 週期變化,所有數據進行統計學分析.結果:p21waf1轉染組 p21waf1 mRNA 和 p21蛋白高錶達(P<0.01);p21waf1轉染組BGC-823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轉染組細胞的 G1期細胞比例顯著高于空載體組和未轉染組(P<0.01),S 期比例顯著低于空載體組和未轉染組(P<0.01),併齣現凋亡峰.結論:p21waf1基因轉染可以抑製人胃癌細胞繫BGC-823細胞增殖併能誘導其髮生細胞凋亡,可能為胃癌的基因治療提供一條新的途徑.
목적:탐토외원성 p21waf1기인전염대인위암세포계 BGC-823세포증식적영향.방법:용지질체개도적방법장 pIRES-p21waf1진핵표체재체전염인위암세포계 BGC-823,분위 pIRES-p21waf1전염조、pIRES-neo 공재체조급미전염조삼개조별.응용실시형광정량 RT-PCR、Western blot 면역인적분별재기인화단백량개수평검측각조 p21적표체정황,채용사갑기우담서람(MTT)법측정세포증식능력,류식세포의분석세포 DNA 주기변화,소유수거진행통계학분석.결과:p21waf1전염조 p21waf1 mRNA 화 p21단백고표체(P<0.01);p21waf1전염조BGC-823세포생장속도명현저우공재체조화미전염조(P<0.01);류식세포의관찰도 p21waf1전염조세포적 G1기세포비례현저고우공재체조화미전염조(P<0.01),S 기비례현저저우공재체조화미전염조(P<0.01),병출현조망봉.결론:p21waf1기인전염가이억제인위암세포계BGC-823세포증식병능유도기발생세포조망,가능위위암적기인치료제공일조신적도경.
Objective: To explore the role of transfected p21waf1 gene in the proliferation of human gastric carcinoma cells.Method: p21waf1 gene packaged with lipofectin was transfected into the cells of a human gastric carcinoma cell line BGC-823,which stably expressed p21waf1 gene.The p21waf1 mRNA and protein expression of BGC-823-p21waf1,and cells were detected by real time PCR and western blot methods,respectively.The cell growth was detected by using MTT.The cell cycle pattern was assayed by using flow cytometry.Result: After transfection of pIRES-p21waf1 The p21waf1 mRNA and protein of BGC-823-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,and that of the cells in S phases was obviously decreased. respectively,as compared with control-group.Conclusion: p21waf1 gene can inhibit the the growth and induce apoptosis of human gastric carcinoma cells, that could be a new method by gene therapy to treat the gastric carcinoma in future.
