摘要
目的探讨透明质酸合成酶2(HAS2)在肾细胞癌(RCC)中的作用及其潜在分子机制。方法收集我院从2019年8月至2021年7月收治的RCC患者接受手术切除的30例RCC组织(RCC组)和30例癌旁组织(NAT组),RT-qPCR和Western blot实验分别检测HAS2 mRNA和蛋白在RCC组织和细胞中的表达。将NC-siRNA、HAS2-siRNA转染至细胞后,细胞计数试剂盒-8(CCK-8)检测RCC细胞的活力;流式细胞术检测RCC细胞凋亡情况;细胞划痕实验检测细胞迁移情况;Western blot检测RCC细胞中上皮-间充质转化(EMT)标志物E-cadherin、N-cadherin、Vimentin蛋白和凋亡蛋白标志物Bax、Caspase-3、Bcl-2蛋白以及TGF-β1、p-Smad4蛋白的表达。结果与NAT组比较,RCC组织中HAS2 mRNA和蛋白水平均显著升高(P<0.05);与正常肾近端小管上皮细胞(RPTEC)比较,RCC细胞系中HAS2 mRNA和蛋白水平均显著升高(P<0.05)。与Control组比较,HAS2-siRNA组细胞中HAS2 mRNA和蛋白水平显著降低,细胞增殖和迁移能力显著降低,细胞凋亡率显著升高,Bax和Caspase-3蛋白水平显著升高,Bcl-2蛋白水平显著降低,E-cadherin蛋白水平显著升高,N-cadherin和Vimentin蛋白水平显著降低,TGF-β1、p-Smad4蛋白水平均显著降低,差异均有统计学意义(P<0.05)。NC-siRNA组与Control组比较差异无统计学意义(P>0.05)。结论沉默HAS2能抑制肾细胞癌细胞增殖、迁移和EMT,促进肾细胞癌细胞凋亡,其机制可能与调控TGF-β/SMAD4信号通路有关。
Objective To investigate the role and potential molecular mechanism of hyaluronic acid synthase 2(HAS2)in renal cell carcinoma(RCC).Methods The cancer tissues(RCC group)and adjacent tissues(NAT group)of RCC patients undergoing surgical resection in our hospital were collected,and RT-qPCR and Western blot were used to detect the expression of HAS2 mRNA and protein in RCC tissues and cells,respectively.After the NC-siRNA and HAS2-siRNA plasmids were transfected into the cells,the cell counting kit-8(CCK-8)was used to detect the viability of RCC cells.The flow cytometry was used to detect the apoptosis of RCC cells.Cell scratch assay was used to detect cell migration.Western blot was used to detect the expressions of epithelial-mesenchymal transition(EMT)markers E-cadherin,N-cadherin,Vimentin protein and apoptosis protein markers Bax,Caspase-3,Bcl-2 and TGF-β1,p-SMAD4 proteins in renal carcinoma cells.Results Compared with the NAT group,HAS2 mRNA and protein levels were significantly increased in RCC tissues(P<0.05);compared with normal renal proximal tubular epithelial cells(RPTEC),HAS2 mRNA and protein levels were significantly increased in RCC cell lines(P<0.05).Compared with the control group,the HAS2 mRNA and protein levels were significantly decreased,the cell proliferation and migration ability were significantly reduced,the apoptosis rate was significantly increased,and the protein levels of Bax and Caspase-3 were significantly increased in the HAS2-siRNA group.The protein level of Bcl-2 was significantly decreased,the protein level of E-cadherin was significantly increased,the protein levels of N-cadherin and Vimentin were significantly decreased,and the protein levels of TGF-β1 and p-Smad4 were significantly decreased in the HAS2-siRNA group,and the differences were statistically significant(P<0.05).There was no significant difference between the NC-siRNA group and the Control group(P>0.05).Conclusion Silencing HAS2 could inhibit the proliferation,migration and EMT of renal cell carcinoma cells,and promote the apoptosis of renal cell carcinoma cells,which might be related to the regulation of TGF-β/SMAD4 signaling pathway.
作者
刘军
冉灿
曾定胜
奉友刚
Liu Jun;Ran Can;Zeng Dingsheng;Feng Yougang(Department of Urology,Suining Central Hospital,Suining Sichuan 629000,China)
出处
《遵义医科大学学报》
2023年第4期396-402,共7页
Journal of Zunyi Medical University
基金
四川省医学科研计划项目(NO:S16077)
超声医学工程国家重点实验室开放课题(NO:2021KFKT018)。
