【摘要】 目的:研究卡波济肉瘤相关疱疹病毒(KSHV)是否能够感染前列腺非致瘤性上皮细胞系RWPE-1及前列腺癌细胞系PC-3,并初步探索其相关信号通路变化。方法:将12-O-十四烷酰佛波醇-13-乙酯(TPA)刺激人原发性渗出性淋巴瘤细胞系BC-3,收集细胞上清(含KSHV病毒颗粒),感染RWPE-1及PC-3,观察前列腺细胞的细胞病变效应(CPE)。采用RT-PCR检测KSHV即刻基因ORF50 mRNA转录状况;进一步运用Western blot检查病毒裂解期产物病毒白细胞介素6(vIL-6)的蛋白表达水平;最后采用Western blot法检测被感染细胞胞浆和胞核中β-catenin表达水平。结果:PC-3细胞感染KSHV后未见明显CPE,但RT-PCR和Western blot均在预期位置检测到KSHV基因ORF50和vIL-6的mRNA及其编码的蛋白条带。RWPE-1细胞感染KSHV后可出现明显的CPE,感染后24h可以检测到vIL-6的表达。Wnt通路蛋白β-catenin在感染KSHV的RWPE-1细胞胞浆和胞核均呈上升趋势。结论:KSHV可以感染前列腺细胞系并有可能激活Wnt通路。更多还原

【Abstract】 Objective:To certify that KSHV could infect prostate non-tumorigenic cells RWPE-1 and prostate cancer cells PC-3 effectively,and to investigate the alteration of the Wnt signalling pathway post-infection. Methods:Harvested from primary effusion lymphoma (PEL) cells BC-3,the KSHV virion- supernatant was used to infect RWPE-1 and PC-3 cells. The cytopathic effect (CPE) of prostate cells was observed post-infection and the replication of KSHV in prostate cells was detected by RT-PCR and Western blot. The expression of β-catenin in both cytoplasm and nuclei was detected by Western blot. Results:Post-infection,CPE was appeared in RWPE-1 but not in PC-3. KSHV ORF50 mRNA was detected in PC-3 cells by RT-PCR. And the specific band of vIL-6 appeared as expected in Western blot detection in both RWPE-1 and PC-3 cells extract. The expression of β-catenin in both cytoplasm and nuclei of RWPE-1 were up-regulated post-infection. Conclusion:KSHV could infect prostate non- tumorigenic cells RWPE-1 and prostate cancer cells PC-3 effectively and trigger the Wnt signaling pathway in RWPE-1 post-infection.更多还原