摘要
目的观测p53调节的凋亡诱导蛋白1(p53AIP1)对PC-3M人前列腺癌细胞的增殖、细胞周期、凋亡、侵袭及迁移的影响。方法构建真核载体pDC316-p53AIP1,并进行双酶切及PCR鉴定。在LipofectamineTM2000介导下转染至体外培养的PC-3M人前列腺癌细胞系中,Western blot法检测其表达,CCK-8法检测PC-3M细胞增殖情况,流式细胞术分析PC-3M细胞周期变化和annexinV-FITC/PI结合流式细胞术凋亡情况,TranswellTM实验检测p53AIP1对细胞迁移及侵袭能力的影响。结果成功构建了pDC316-p53AIP1真核表达载体,重组质粒转染PC-3M细胞后,Western blot法检测p53AIP1在PC-3M细胞中有表达,CCK-8结果显示表达的p53AIP1能抑制PC-3M细胞增殖(P<0.05),流式细胞术检测细胞周期阻滞在S/G2-M期(P<0.05)且p53AIP1能促进PC-3M细胞凋亡,TranswellTM侵袭和迁移实验显示p53AIP1使细胞侵袭迁移能力均降低(P<0.05)。结论p53AIP1抑制PC-3M细胞的周期、增殖、侵袭及迁移,并能促进其凋亡。
Objective To investigate the effect of p53-regulated apoptosis-inducing protein 1 （p53AIP1） gene on the proliferation, cell cycle, apoptosis, invasion and migration of PC-3M human prostate cancer cells in vitro. Methods The eukaryotic expression vector pDC316-p53AIP1 was constructed and confirmed by double enzyme digestion and PCR analysis. Then it was transfected into PC-3M human prostate cancer cells by LipofectamineTM 2000. The expression of p53AIP1 protein was detected by Western blotting. The proliferation of PC-3M cells was tested by CCK-8 assay; the cell cycle and apoptosis rate were analyzed by flow cytometry combined with annexin V-FITC/PI staining; the effect of p53AIP1 on the cell invasion and migration was detected by TranswellTM assay. Results The eukaryotic expression vector pDC316-p53AIP1 was constructed successfully. After transfected into PC-3M cells, Western blotting demonstrated that the protein p53AIP1 was effectively expressed. CCK-8 assay revealed that the proliferation of PC-3M cells was significantly inhibited by p53AIP1（P〈0.05）. Flow cytometry indicated that the cells were arrested at S/G2-M phase （P 〈 0.05） and cell apoptosis was evidently promoted （P 〈0.05）. TranswellTM chamber experiments showed that p53AIP1 decreased the abilities of both invasion and migration of the cells （P〈0.05）. Conclusion The p53AIP1 inhibits the proliferation of PC-3M cells, arrests cellcycle at S/G2-M phase, decreases the abilities of invasion and migration and promotes cell apoptosis.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第8期819-823,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81201612)
国家高技术研究发展计划(863)(2007AA02Z451)
