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背景与目的:PAR(prostate androgen regulated)基因在雄激素非依赖性的前列腺癌中特异性高表达,可能与细胞的恶性转化有关。本研究采用RNA干扰技术下调前列腺癌PC3细胞中PAR基因的表达,并探讨其对PC3细胞恶性表型的影响及作用机制。方法:构建针对PAR基因的短发夹RNA(short hairpin RNA,shRNA)表达质粒psiRNA-PAR1、psiRNA-PAR2和psiRNA-PAR3,转染PC3细胞;采用RT-PCR法检测其对PAR基因的表达抑制作用;细胞计数、软琼脂克隆形成实验、流式细胞术检测PAR基因表达下调对PC3细胞生长的抑制作用及其作用机制。结果:3组shRNA表达质粒均可抑制PC3细胞中PAR基因的表达;PAR基因的下调可使细胞生长速率明显减慢,增殖受到抑制;克隆形成能力降低,其中以psiRNA-PAR1的抑制效应最明显,并在转染48h后对PAR基因表达的抑制率达最高峰,为(81.18±1.68)%;流式细胞术检测结果表明,细胞周期阻滞于G2/M期,凋亡增加。结论:PAR基因表达下调,明显地抑制PC3细胞的生长,其作用机制主要是通过诱导细胞G2/M期阻滞和凋亡实现的;PAR基因可能是一个新的与雄激素非依赖性前列腺癌细胞恶性表型相关的癌基因,有望作为基因及药物治疗雄激素非依赖性前列腺癌的一个新靶点。
BACKGROUND & AIM: PAR (prostate androgen regulated) genes are highly expressed in androgen-independent prostate cancer, which may be related to the malignant transformation of cells. In this study, RNAi was used to down-regulate the expression of PAR gene in prostate cancer PC3 cells and to explore its effect on the malignant phenotype of PC3 cells and its mechanism. Methods: The shRNA expression plasmids psiRNA-PAR1, psiRNA-PAR2 and psiRNA-PAR3 for PAR gene were constructed and transfected into PC3 cells. The expression of PAR gene was detected by RT-PCR The cell counting, soft agar colony formation assay and flow cytometry were used to detect the inhibitory effect of PAR gene expression on PC3 cell growth and its mechanism. Results: The shRNA expression plasmid of all three groups could inhibit the expression of PAR gene in PC3 cells. The down-regulation of PAR gene could significantly slow down the cell growth and inhibit the proliferation, and the ability of clonality decreased. The inhibitory effect of psiRNA-PAR1 was the most obvious (81.18 ± 1.68)%. The results of flow cytometry showed that cell cycle arrest in G2 / M phase and apoptosis increased. CONCLUSION: Down-regulation of PAR gene significantly inhibits the growth of PC3 cells, and its mechanism is mainly through induction of G2 / M arrest and apoptosis. PAR gene may be a new androgen independent prostate cancer The malignant phenotype related oncogene is expected to serve as a new target of gene and drug therapy for androgen independent prostate cancer.