腺病毒介导的PTEN和p16基因对前列腺癌细胞系的作用  

Effect of PTEN and p16 Expression in Prostate Cancer PC-3 Cell

作　　者：崔飞伦[1] 邱镇 于洋[3] 姚震[1] 林大春[1] 

机构地区：[1]江苏省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 [3]辽宁医学院基础医学系,辽宁锦州121000

出　　处：《医学临床研究》2007年第3期373-376,共4页Journal of Clinical Research

摘　　要：【目的】探讨腺病毒介导的PTEN和p16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PTEN和p16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-PTEN为1.8×107pfu/ml、Ad-p16为1.2×1010pfu/ml,RT-PCR检测可见PTEN-mRNA(462bp)和p16-mRNA(520bp)表达,Western blot检测有PTEN蛋白(60KD)和p16蛋白(16KD)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。【结论】PTEN和p16可明显的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PTEN和p16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 effect of PTEN and p16 gene expression on the cell proliferation and apoptosis of prostate cancer PC-3 cell. [MethodslRecombinant adenovirus vector of human tumor suppressor gene PTEN and p16 was constructed and infected prostate cancer PC-3 cell line in vitro, the mRNA and protein expressions of PTEN and p16 in prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-p16 were determined by RT-PCR and Western blot respectively. MTT assay was used to determine the effect of PTEN and p16 on proliferation of PC-3 cell,the apoptosis rate of PC-3 cell was examined by flow cytometry. [Results]The viral titers of Ad-PTEN and Ad-p16 were 1.8 ×10^7 pfu/ml, 1.2 ×10^10pfu/ml respectively. After infected by adenovirus ,it had been verified that the mRNA and protein expression of PTEN and p16 were steady in human PC-3 cell, Ad-PTEN and Ad-p16 inhibited the growth and proliferation of PC-3 cells, The progression of cell cycle of PC-3 cell was arrested in G0-G1 phase ,meanwhile the apoptosis rate of PC-3 was also affected after Ad-PTEN or/and Ad-p16 infected. There was significant difference between combined therapy group and single gene therapy group, [Conclusion]The recombinant Ad-PTEN and Ad-pl6 vectors are constructed successfully and the expression of specific PTEN and p16 is high , steady in PC-3 cell line, These resuits suggest that combination of PTEN and p16 has application value in treatment of prostate cancer in future.

关 键 词：前列腺肿瘤 腺病毒科 基因 抑制 肿瘤 基因 p16 

分 类 号：R737.25[医药卫生—肿瘤]