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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 observe the effect of p53-regulated apoptosis-inducing protein 1 (p53AIP1) on the proliferation, cell cycle, apoptosis, invasion and migration of PC-3M human prostate cancer cells. Methods The eukaryotic vector pDC316-p53AIP1 was constructed and double digested and identified by PCR. PC-3M human prostate cancer cell lines transfected with LipofectamineTM2000 were detected by Western blot, CCK-8 assay PC-3M cells proliferation, PC-3M cell cycle analysis by flow cytometry Changes and annexinV-FITC / PI combined with flow cytometry apoptosis, TranswellTM experiment to detect p53AIP1 cell migration and invasion ability. Results The eukaryotic expression vector pDC316-p53AIP1 was constructed successfully. The expression of p53AIP1 was detected in PC-3M cells by Western blot and PCA-3M, Proliferation (P <0.05). Flow cytometry showed that cell cycle arrest was at S / G2-M phase (P <0.05) and p53AIP1 could promote PC-3M cell apoptosis. TranswellTM invasion and migration assays showed that p53AIP1 caused cell invasion and migration Ability decreased (P <0.05). Conclusion p53AIP1 can inhibit the cell cycle, proliferation, invasion and migration of PC-3M cells and promote its apoptosis.