HSPC111异常表达对前列腺癌细胞生物学行为的影响

发布时间：2019-03-15 07:29

【摘要】：目的：明确HSPC111在前列腺癌中的表达情况及其对前列腺癌细胞体外生物学行为的影响,探讨HSPC111在前列腺癌进展中的作用。方法：运用RT-qPCR方法检测HSPC111在前列腺癌和前列腺增生组织中的表达量；利用RT-qPCR及Western Blot检测HSPC111在正常前列腺细胞系RWPE-1以及前列腺癌细胞系LNCaP和PC3中的表达量；构建含HSPC111短发卡RNA(shRNA)的慢病毒载体,转染PC3细胞,经嘌呤霉素筛选及挑选单克隆后获得稳定低表达HSPC111的PC3细胞株,并利用RT-qPCR及Western Blot验证转染后细胞中HSPC111的表达量；运用细胞计数法和MTT法检测HSPC111低表达PC3细胞的增殖能力,利用Transwell法检测HSPC111低表达PC3细胞侵袭能力,磷脂酰丝氨酸外翻分析(Annexin V)检测HSPC111低表达PC3细胞的凋亡情况；通过测定细胞内蛋白含量检测HSPC111异常表达对细胞蛋白合成的影响。所有实验数据均采用SPSS18.0软件进行统计学分析。结果：与前列腺增生组织相比,HSPC111在前列腺癌组织中高表达；与正常前列腺细胞系RWPE-1相比,HSPC111在前列腺癌细胞系LNCaP及PC3中高表达,而LNCaP与PC3之间无明显差异；利用含HSPC111特异性shRNA的慢病毒载体转染PC3细胞可建立HSPC111稳定低表达的细胞；降低内源性HSPC111表达可抑制细胞体外增殖；降低内源性HSPC111表达可抑制细胞侵袭能力；降低内源性HSPC111表达可促进细胞凋亡；降低内源性HSPC111表达可抑制细胞蛋白合成。结论： HSPC111异常表达可改变前列腺癌细胞的体外生物学行为,且与前列腺癌进展密切相关。我们认为抑制HSPC111表达对控制前列腺癌增殖进展是一个可行的治疗策略。
[Abstract]:Aim: to investigate the expression of HSPC111 in prostate cancer and its effect on the biological behavior of prostate cancer cells in vitro, and to explore the role of HSPC111 in the progression of prostate cancer. Methods: RT-qPCR method was used to detect the expression of HSPC111 in prostate cancer and benign prostatic hyperplasia, RT-qPCR and Western Blot were used to detect the expression of HSPC111 in normal prostate cell line RWPE-1 and prostate cancer cell line LNCaP and PC3. The lentivirus vector containing HSPC111 short hairpin RNA (shRNA) was constructed and transfected into PC3 cells. PC3 cell lines with low expression of HSPC111 were obtained by puromycin selection and monoclonal selection. The expression of HSPC111 in transfected cells was verified by RT-qPCR and Western Blot. The proliferative ability of PC3 cells with low expression of HSPC111 was detected by cell counting method and MTT method, the invasive ability of PC3 cells with low expression of HSPC111 was detected by Transwell method, and the apoptosis of PC3 cells with low expression of HSPC111 by (Annexin V) was detected by phosphatidyl serine valgus analysis. The effect of abnormal expression of HSPC111 on cell protein synthesis was detected by measuring intracellular protein content. All the experimental data were analyzed by SPSS18.0 software. Results: compared with benign prostatic hyperplasia, HSPC111 was highly expressed in prostate cancer tissue, HSPC111 was higher in LNCaP and PC3 than that in normal prostate cell line RWPE-1, but there was no significant difference between LNCaP and PC3. PC3 cells were transfected with lentiviral vector containing HSPC111-specific shRNA to establish stable and low-expression cells of HSPC111, lower expression of endogenous HSPC111 could inhibit cell proliferation in vitro, lower expression of endogenous HSPC111 could inhibit invasion ability of cells, and lower expression of endogenous HSPC111 could inhibit invasion ability of cells. Down-regulation of endogenous HSPC111 expression can promote cell apoptosis and decrease endogenous HSPC111 expression can inhibit cell protein synthesis. Conclusion: the abnormal expression of HSPC111 can change the biological behavior of prostate cancer cells in vitro, and it is closely related to the progression of prostate cancer. We believe that the inhibition of HSPC111 expression is a feasible therapeutic strategy to control the progression of prostate cancer proliferation.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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