摘要
目的:探讨p21WAF1(p21)基因转染对人胰腺癌细胞系BxPC-3细胞增殖的影响.方法:根据转染质粒的不同和是否进行质粒转染分为3组,p21转染组、空载体转染组和未转染组.应用RT-PCR和Western blot方法检测转染细胞的p21基因表达变化;流式细胞仪分析细胞周期变化,用MTT、流式细胞仪和透射电镜检测转染外源p21基因对BxPC-3细胞增殖和凋亡的影响.结果:p21转染组细胞存在p21 mRNA高表达和P21蛋白高表达;p21转染组细胞生长速度低于对照空载体转染组和未转染组;流式细胞仪观察到P21蛋白高表达使BxPC-3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(59.887%±3.700% vs 47.443%±6.354%,49.223%±2.226%,P<0.05),S期显著低于空载体组和未转染组(21.277%±2.080% vs 35.247%±3.966%,36.013%±1.540%,P<0.01),并出现亚G1峰(凋亡峰).透射电镜亦发现p21转染组发生细胞凋亡.结论:p21基因转染可以抑制人胰腺癌细胞系BxPC-3细胞增殖并能诱导其发生细胞凋亡.
AIM： To investigate the effect of p21WAF1 （p21） gene transfection on the proliferation of humanpancreatic cancer line BxPC-3 METHODS： According to the different trans- fected plasmid and whether the transfection was performed, 3 groups were formed. For p21 transfection group, pCDNA3.1 （＋）-p21 was trans- fected into BxPC-3 cell by the vector of Lipofectamine2000. For empty plasmid transfection group, pCDNA3.1 （＋）-neo plasmid was transfected into BxPC-3 cell with the same method as the blank control group. For non-transfection group, the BxPC-3 cell was not transfected as the negative control group. The expression of p21 was evaluated using RT-PCR and Western blot. The proliferation and apoptosis were determined by MTT, flow cytometry and transmis- sion electron microscopy. RESULTS： After transfection, the expression of p21 mRNA and P21 protein was up regulated, and the cell growth was decreased. High protein expression of P21 BxPC-3 cell cycle was arrested at G1/S phase, the population of G1 phase was significantly increased, compared with the empty plasmid transfection group or the non-transfection group （59.887% ±3.700% vs 47.443%± 6.354%, 49.223% ±2.226%, P 〈 0.05）, the S phase population was significantly decreased （21.277% ± 2.080% vs 35.247% ± 3.966%, 36.013% ± 1.540%, P 〈 0.01）, and a Sub-G1 peak （apoptosis peak） appeared. Cell apoptosis by transmission electron microscopy was observed in the pCDNA3.1 （＋）-p21 transfection group BxPC-3 cells. CONCLUSION： After p21 gene transfection, BxPC-3 cell proliferation is significantly arrested and apoptosis could be induced in vitro.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第16期1614-1620,共7页
World Chinese Journal of Digestology
基金
黑龙江省科技计划攻关重点项目基金资助项目
No.GB05C401-09
黑龙江省教育厅科学技术研究基金资助项目
No.11521191
黑龙江省卫生厅科研基金资助项目
No.2007-304~~
