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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。
Objective: To investigate whether Kaposi’s sarcoma-associated herpes virus (KSHV) can infect prostate epithelial non-tumorigenic epithelial cell line RWPE-1 and prostate cancer cell line PC-3, and to explore its related signaling pathways. Methods: Human primary exudative lymphoma cell line BC-3 was stimulated with 12-O-tetradecanoylphorbol-13-ethyl ester (TPA) and cell supernatants (including KSHV virus particles) RWPE-1 and PC-3 were used to observe the cytopathic effect (CPE) of prostate cells. RT-PCR was used to detect the transcriptional status of ORF50 mRNA in KSHV-immediate gene. Western blot was used to detect the protein expression level of virus IL-6 in virus lysates. Finally, Nuclear β-catenin expression levels. RESULTS: No significant CPE was detected in PC-3 cells infected with KSHV. However, both mRNA and protein coding regions of ORF50 and vIL-6 of KSHV gene were detected at expected positions by RT-PCR and Western blot. Significant CPE occurred in RWPE-1 cells infected with KSHV, and the expression of vIL-6 was detected 24h after infection. The Wnt pathway protein β-catenin showed an upward trend in the cytoplasm and nucleus of RWPE-1 cells infected with KSHV. Conclusion: KSHV can infect prostate cell lines and possibly activate Wnt pathway.