p16和p27表达对前列腺癌细胞系的作用  

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

作　　者：崔飞伦[1] 邱镇 陆洪兵[1] 姚震[1] 林大春[1] 

机构地区：[1]镇江市第一人民医院泌尿外科,江苏镇江212002 [2]镇江359医院泌尿外科,江苏镇江212000

出　　处：《解剖科学进展》2007年第1期18-20,共3页Progress of Anatomical Sciences

摘　　要：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115×108 pfuPml、Ad-p27为112×109 pfuPml,RT-PCR检测可见p16-mRNA(520bp)和p27Kip12 mRNA(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。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 ×10^8 pfuPml and 112 × 10^9 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-pl6 and Ad-p27 inhibited the growth and proliferation of PC-3 cells, the progress of cell cycle of PC-3 cell was arrested in GO -GI phase, meanwhile the apoptosis rate of PC-3 was also increased with significant difference between combined therapy group and single gene therapy group. Coildusion The recombinant Ad-pl6 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.

关 键 词：前列腺肿瘤 基因 腺病毒 治疗 

分 类 号：R737.25[医药卫生—肿瘤]