PBF基因对前列腺癌细胞PC3增殖和侵袭的影响

Effect of PBF gene on proliferation and invasion of prostate cancer cell line PC3

目的探讨沉默垂体肿瘤转化基因结合因子(PBF)基因的表达对前列腺癌细胞PC3增殖、侵袭及化疗药物耐药的影响。方法利用慢病毒转染法将靶向PBF干扰质粒转染前列腺癌细胞PC3,Western blot检测质粒的转染效果;噻唑蓝(MTT)和软琼脂克隆形成实验检测各组细胞的增殖及成瘤能力的变化;流式细胞术检测PBF沉默前后PC3细胞周期的变化;Transwell实验和划痕实验检测细胞的侵袭和迁移能力;细胞存活率实验检测细胞对化疗药物的耐药性。结果PBF干扰质粒能降低前列腺癌细胞PC3中PBF的表达;沉默PC3细胞中PBF表达后,与对照组比较,PC3/shPBF细胞增殖能力明显降低(P=0.002)且细胞周期阻滞于G 1期(P=0.046),PC3/shBPF细胞成瘤能力明显降低(P=0.000);PBF表达被沉默后,PC3细胞侵袭转移能力下降(P=0.001);PC3细胞PBF表达降低后,对前列腺癌常用化疗药物紫杉醇的敏感性增加(P=0.013)。结论沉默PBF的表达能降低前列腺癌细胞PC3的增殖和侵袭能力并增强其对化疗药物的敏感性,提示PBF可能会成为前列腺癌治疗的潜在靶点。

Objective To investigate the effect of silencing PTTG binding factor(PBF)gene expression on the proliferation,invasion and chemotherapy resistance of prostate cancer cell line PC3.Methods The lentivirus transfection method was used to transfect the interfering plasmid of targeted PBF to prostate cancer cell line PC3.The effect of plasmid was detected by Western blot.MTT and soft agar clone formation experiment were used to detect the cell proliferation and change of tumorigenesis ability.The change of PC cell cycle before and after PBF silence was detected by the fluid cytometry.The ability of cellular invasion and migration was detected by the Transwell experiment and scratch experiment.The cellular viability test was used to detect the resistance of cell to chemotherpeutic drugs.Results PBF interfering the plasmid could reduce the PBF expression level in PC3 cells;after silencing PBF expression in P3,MTT experiment and fluid cytometry indicated that compared with the control group,the cell proliferation rate was significantly decreased(P=0.002),moreover the cellular cycle was arrested at the G 1 stage(P=0.046).In the soft agar clone formation experiment,after silencing PBF expression,the tumorigenesis ability was significantly decreased(P=0.000).Transwell experiment and scratch experiment showed that after silencing PBF expression,the invasion and migration ability of PC3 was decreased(P=0.001).After PBF expression decrease in P3 cells,its sensitivty to ehemotherapeutic drug paclitaxel in prostate cancer was increased(P=0.013).Conclusion Silencing PBF expression can decrease the proliferation and invasion ability of P3 cells,enhance its sensitivity to chemotherapeutic drug,indicating that PBF may become the latent target spot of prostate cancer therapy.