摘要
垂体肿瘤转化基因1(PTTG1)具有促进肿瘤生长和转移的作用.通过上调或下调基因表达的策略,观察PTTG1基因对人前列腺癌细胞株LNCaP细胞生长增殖的影响.利用PCR技术分离出PTTG1全长cDNA,分别正向和反向插入真核表达载体pIRES2-EGFP,重组载体分别命名为正义PTTG1-S/pIRES2-EGFP(即pI-P-S)和反义PTTG1-AS/pIRES2-EGFP(即pI-P-AS),将这两种重组载体稳定转染LNCaP细胞,通过流式细胞仪和MTT法分别检测了细胞周期和细胞增殖的情况.转染正义PTTG1后处于S期和G2期的细胞明显增加,细胞生长增殖能力增强;相反,转染反义PTTG1后处于S期和G2期细胞明显减少,细胞生长增殖能力减弱(P<0.05).结果表明,PTTG1能明显改变人前列腺癌细胞株LNCaP的细胞周期和细胞生长增殖能力,它的异常表达可能参与前列腺癌细胞生长增殖过程.
Pituitary tumor transforming gene 1 （PTTG1） is a proto-oncogene and plays roles in promoting tumorigenesis and metastasis. Eukaryotic expression vector expressing full-length sense and anti-sense PTTG1 mRNA and their effect on the biological behavior of human prostate cancer cell llne LNCaP were observed. The full-length PTTG1 eDNA fragment was inserted into the eukaryotic vector pIRES2-EGFP in the sense and antisense direction, named PTTG 1-S/pIRES2- EGFP （pI-P-S） and PTTG 1-AS/pIRES2- EGFP （pI-P-AS） . The two recombinant vectors were transfected into LNCaP by LipofectAMINE2000 respectively. The LNCaP clones stably expressing full-length recombinant PTTG1-S/pIRES2-EGFP（sense） or PTTG1-AS/pIRES2-EGFP （antisense） were screened with G418.The changes of cell cycle progression and proliferation were analyzed by flow cytometry and MTT methods, respectively. The cell cycle progression was increased during S and peaked at G2 phase, and cell proliferation was accelerated in sense-transfected cells. In contrast the cell cycle progression was decreased during S and G2 phase, and cell proliferation was reduced in anti-sense-transfected cells as compared with control cells, （ P 〈 0.05）. The results indicated that PTTG1 may change the cell cycle progression and improve the potential of cell proliferation in LNCaP cell. These data also suggested that PTTG1 has potential to play an important role in the proliferation of prostate cancer cell.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2005年第5期679-684,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.30171027)资助~~
关键词
垂体瘤转化基因1
前列腺肿瘤
细胞增殖
pituitary tumor transforming genel （ PTTG1 ）, prostate cancer, cell proliferation
