摘要
目的研究lncRNAFGD5-AS1对肾癌细胞增殖、侵袭的影响及分子机制。方法2020年7月至2021年2月,采用实时荧光定量聚合酶链式反应(qRT-PCR)检测FGD5-AS1在肾癌细胞中的表达。以表达最高的肾癌细胞为对象,分别转染小干扰siRNA-FGD5-AS1质粒(实验组)和阴性对照质粒(对照组)。qRT-PCR检测FGD5-AS1和细胞因子信号转导抑制因子6(suppressorof cytokinesignaling 6,SOCS6)mRNA的表达。Westernblotting检测SOCS6和NF-κB信号通路蛋白表达。二苯基溴化四氮唑蓝(MTT)实验和Transwell侵袭实验检测细胞增殖和侵袭。结果肾癌细胞株中FGD5-AS1表达均高于近端肾小管细胞(P<0.01),FGD5-AS1在ACHN细胞中表达最多(P<0.01)。对照组和实验组FGD5-AS1表达分别为(1.01±0.09)和(0.20±0.03),差异有统计学意义(t=8.46,P<0.01);SOCS6 mRNA的表达分别为(0.33±0.05)和(1.02±0.13),差异有统计学意义(t=5.05,P<0.01)。与对照组相比,实验组SOCS6蛋白表达增加,NF-κB信号通路蛋白表达均明显降低,实验组ACHN细胞增殖能力降低(均P<0.05)。对照组和实验组的侵袭细胞数分别为(80.49±10.50)个和(32.29±8.03)个,差异有统计学意义(t=3.65,P<0.05)。结论FGD5-AS1在肾癌细胞株中表达增加,下调FGD5-AS1表达可通过正向调控SOCS6基因表达抑制肾癌细胞的增殖和侵袭。
Objective To study the effect of lncRNA FGD5-AS1 on the proliferation and invasion of renal cancer cells and the molecular mechanism.Methods From July,2020 to February,2021,real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the FGD5-AS1 expression in the renal cancer cells.The highest expression renal cancer cells were used as the objects.The small interfering siRNA-FGD5-AS1 plasmid(an experimental group)and the negative control plasmid(a control group)were transfected.QRT-PCR was used to detect the mRNA expressions of FGD5-AS1 and the suppressor of cytokine signaling 6(SOCS6).Western blotting was used to detect the expressions of SOCS6 and NFκB signaling pathway proteins.Methylthiazolyldiphenyl-tetrazolium bromide(MTT)test and Transwell invasion test were used to detect the cell proliferation and invasion.Results The expression of FGD5-AS1 in the renal cancer cell lines was higher than that in the proximal renal tubular cells(P<0.01),and FGD5-AS1 was the most expressed in the ACHN cells(P<0.01).The expressions of FGD5-AS1 in the control group and the experimental group were(1.01±0.09)and(0.20±0.03)(t=8.46,P<0.01).The expressions of SOCS6 mRNA in the control group and the experimental group were(0.33±0.05)and(1.02±0.13)(t=5.05,P<0.01).Compared with those in the control group,the SOCS6 protein was significantly increased and the NFκB signaling pathway protein was significantly reduced in the experimental group,and the proliferation of the ACHN cells in the experimental group was significantly reduced(all P<0.05).The numbers of the invaded cells in the control group and the experimental group were(80.49±10.50)and(32.29±8.03)(t=3.65,P<0.05).Conclusions FGD5-AS1 is significantly increased in renal cancer cell lines.Down-regulating FGD5-AS1 expression can inhibit the proliferation and invasion of renal cancer cells by positively regulating SOCS6 gene expression.
作者
叶志华
付金伦
桂定文
罗帅
王静
王小英
Ye Zhihua;Fu Jinlun;Gui Dingwen;Luo Shuai;Wang Jing;Wang Xiaoying(Department of Urological Surgery,Huangshi Central Hospital,Edong Healthcare Group(Affiliated Hospital of Hubei Polytechnic University),Hubei Key Laboratory of Kidney Disease Pathogenesis and Intervention,Huangshi 435000,China;Department of Surgery,Huangshi Second Hospital,Edong Healthcare Group,Huangshi 435000,China)
出处
《国际医药卫生导报》
2021年第19期2969-2972,共4页
International Medicine and Health Guidance News
基金
湖北省卫生健康科研基金资助项目(WJ2021Q006)。
