miR-485-5p在卡波西肉瘤和相关疱疹病毒感染患者血清及病毒感染细胞中的表达及生物学功能

The Expression of miR-485-5p in Serum of KS and KSHV Infected Patients, and in Cells Infected with KSHV and its Biological Function

目的 研究miR-485-5p在卡波西肉瘤(Kaposi′s sarcoma, KS)和卡波西肉瘤相关疱疹病毒(KSHV)感染患者血清及KSHV感染细胞中的表达及其对细胞生物学功能的影响。方法 dbDEMC数据库分析miR-485-5p在肉瘤组织中的表达。qRT-PCR分别检测miR-485-5p在KS和KSHV感染患者血清及KSHV感染细胞中的表达水平。miR-485-5p模拟物(miR-485-5p mimic)、miR-485-5p抑制剂(miR-485-5p inhibitor)及对照分别转染到iSLK-219细胞中,CCK-8法和平板克隆检测细胞增殖能力,划痕愈合及transwell实验检测细胞迁移侵袭能力。同时通过Western blot检测EMT(epithelial-mesenchymal transition, EMT)相关蛋白的表达。结果 miR-485-5p在KS和KSHV感染患者血清及KSHV感染细胞中均高表达(P<0.05)。iSLK-219细胞中转染miR-485-5p模拟物后细胞的增殖及迁移侵袭能力增强,而转染miR-485-5p抑制剂后细胞的增殖及迁移侵袭能力减弱(P<0.05)。iSLK-219细胞中转染miR-485-5p模拟物后可促进EMT标志物分子的表达,而转染miR-485-5p抑制剂后可抑制EMT标志物分子的表达(P<0.05)。结论 miR-485-5p在KS和KSHV感染患者血清及KSHV感染细胞中均高表达,且可促进细胞的增殖、迁移侵袭及EMT进程,其有可能作为KSHV感染相关疾病的潜在临床诊治靶点。

Objective To investigate the expression of miR-485-5 p in serum of Kaposi′s sarcoma(KS) and KSHV infected patients, and in cells infected with KSHV and its effects on cell biological function.Methods The expression of miR-485-5 p in sarcoma tissues was analyzed by the dbDEMC database. qRT-PCR was used to detect the expression of miR-485-5 p in serum of KS and KSHV infected patients, and in cells infected with KSHV. The miR-485-5 p mimic, miR-485-5 p inhibitor and controls were transfected into iSLK-219 cell respectively, and the proliferation viability of cell was detected by CCK-8 and colony formation assay. Scratch healing and transwell assay were performed to calculate the migration and invasion abilities of cell. Moreover, the expression of EMT-related molecules was detected by Western blot.Results There were high expression of miR-485-5 p in serum of KS and KSHV infected patients, and in cells infected with KSHV(P<0.05). Transfection of miR-485-5 p mimic in iSLK-219 cell could enhance cell proliferation, migration and invasion abilities, while transfection of miR-485-5 p inhibitor could decrease cell proliferation, migration and invasion abilities(P<0.05). In addition, the expression of EMT molecules could be promoted by transfection of miR-485-5 p mimic in iSLK-219 cell, while the expression of EMT molecules could be inhibited by transfection of miR-485-5 p inhibitor(P<0.05).Conclusion miR-485-5 p is highly expressed in serum of KS and KSHV infected patients, and in cells infected with KSHV, also may promote the process of cell proliferation, migration, invasion and EMT, suggesting that miR-485-5 p has the potential to serve as a potential target for clinical diagnosis and treatment of KSHV infection-related diseases.