p16和p27表达对前列腺癌细胞系的作用

目的 探讨p16和p27Kip1 基因蛋白对抑制前列腺癌细胞增殖和调控其凋亡的作用.方法 构建携带人P16和p27基因的腺病毒载体,转染体外培养的前列腺癌细胞系PC-3,采用RT-PCR、Western blot检测目的 基因的表达.通过细胞生长试验、流式细胞仪检测PC-3转染前后细胞增殖和凋亡的变化.结果 病毒滴度Ad-p16为115×10^8 pfuPml 、Ad-p27为112×10^9 pfuPml ,RT-PCR 检测可见p16-mRNA（520bp）和p27Kip12mRNA （320bp）表达,Western blot检测有p16 蛋白（65KD）和P27蛋白（27KD） 特异表达,并可明显抑制PC-3细胞的增殖,诱导其凋亡,联合基因治疗组与单基因组相比差异显著（P＜0.01）.结论 p16和p27可明显抑制前列腺癌细胞株PC-3的增殖,增加细胞的凋亡,转染携带p16和p27的重组腺病毒载体有望成为治疗前列腺癌的有效方法.

Effects of p16 and p27 Expressions on Prostate Cancer PC-3 Cells

Objective To investigate the effects of p16 and p27 gene expressions on the cell proliferation and apoptosis of prostate cancer PC-3 cells.Methods Recombinant adenovirus vectors of human tumor suppressor gene p16 and p27 were constructed and infected into prostate cancer PC-3 cells line in vivo,the expressions of p16 and p27 in prostate cancer PC-3 cell line were determined by RT-PCR and Western blot respectively.MTT assay was used to determine the effects of p16 and p27 on proliferation of PC-3 cells,the apoptosis rate of PC-3 cell was examined by flow cytometry.Results The viral titers of Ad-p16 and Ad-p27 were 115 ×108 pfuPml and 112 ×109 pfuPml respectively.After transfection by adenovirus,it was verified that the mRNA and protein of p16 and p27 were expressed steady in human PC-3 cells.It was also found that Ad-p16 and Ad-p27 inhibited the growth and proliferation of PC-3 cells,the progress of cell cycle of PC-3 cell was arrested in G0-G1 phase,meanwhile the apoptosis rate of PC-3 was also increased with significant difference between combined therapy group and single gene therapy group.Conclusion The recombinant Ad-p16 and Ad-p27 vectors were constructed successfully with the expression of specific p16 and p27 highly and steadily in PC-3 cell line,which suggests that combination of p16 and p27 has an application value in treatment of prostate cancer in future.