目的 研究脂肪细胞对人正常前列腺上皮细胞RWPE-1及人正常前列腺基质永生化细胞WPMY-1的增殖作用及潜在机制.方法 首先成脂诱导后制备脂肪细胞条件培养基(Adi-cm),然后分别将RWPE-1与WPMY-1细胞各自分为对照组与Adi-cm组.油红O染色验证脂肪诱导结果,cck8试剂盒检测细胞增殖与细胞毒性分析,划痕实验检测细胞迁移能力,RT-PCR 与 Western blot 分别检测核因子-κB(NF-κB)、Bcl-2、Bax、IL-1 β、IL-6、TNF-α 的 RNA水平与蛋白质水平的表达差异.结果 Adi-cm组细胞增殖明显增加,细胞迁移未表现出明显的统计学差异,NF-κB及炎症因子IL-1β、IL-6、TNF-α表达明显增加,NF-κB抑制剂能明显抑制Adi-cm培养后的两种前列腺细胞的增殖.结论 脂肪细胞能通过激活NF-κB,释放炎症因子,促进RWPE-1及WPMY-1细胞的增殖.
The role of adipocytes in promoting prostatic cell proliferation through NF-κB signaling pathway
Objective To investigate the effect of adipocytes on the proliferation of human normal prostate epithelial cell(RWPE-1)and human normal prostate stromal immortalized cell(WPMY-1)and its potential mechanism.Methods Adipocyte conditioned medium(Adi-cm)was prepared and collected after lipogenesis induction.RWPE-1 and WPMY-1 cells were divided into the control group and the Adi-cm group,respectively.The results of fat induction were verified by oil red O staining,cell proliferation and cytotoxicity were detected by cck8 kit,cell migration ability was evaluated by scratching assay,RNA and protein levels of NF-κB,Bcl-2,Bax,IL-1β,IL-6 and TNF-α were detected by RT-PCR and western blot.Results Cell proliferation was significantly increased in the Adi-cm group,and no significant difference was found in cell migration.Expressions of NF-κB and inflammatory factors IL-1β,IL-6 and TNF-α were significantly increased in the Adi-cm group.NF-κB inhibitors could significantly inhibit the proliferation of the two types of prostate cells treated with Adi-cm.Conclusion Adipocytes can promote the proliferation of RWPE-1 and WPMY-1 cells by activating NF-κB and releasing inflammatory factors.
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