长链非编码RNA HNF1A-AS1靶向has-miR-1对膀胱癌细胞生长和侵袭的调节作用及机制研究

目的：探究长链非编码RNA HNF1A-AS1通过靶向miR-1对膀胱癌细胞增殖和侵袭的作用。方法：荧光定量检测膀胱癌细胞系和正常泌尿道上皮细胞HNF1A-AS1和miR-1的表达情况；分别过表达或沉默HNF1A-AS1和miR-1，荧光定量检测miR-1的表达情况；荧光素酶报告实验确定HNF1A-AS1与miR-1的靶向关系；分别沉默HNF1A-AS1和miR-1，MTT和侵袭实验检测细胞活性和侵袭能力，免疫印迹检测细胞增殖及侵袭相关蛋白的表达。结果：根据预实验结果选择253J细胞系进行后续实验。HNF1A-AS1过表达可明显抑制miR-1表达(P<0.05)，减弱miR-1 mimic对miR-1表达的促进作用(P<0.05)。沉默HNF1A-AS1可显著促进miR-1表达(P<0.05)，减弱miR-1 inhibitor对miR-1表达的抑制作用(P<0.05)；同时，HNF1A-AS1还可明显减弱带有miR-1片段野生型质粒的荧光素酶活性(P<0.05)；此外，沉默HNF1A-AS1可显著抑制膀胱癌细胞增殖及侵袭(P<0.05)，并且还能显著抑制细胞增殖相关蛋白Ki67、PCNA和侵袭相关蛋白MMP-9、VEGF的表达(P<0.05)；抑制miR-1表达可明显减弱shRNA对癌细胞增殖和侵袭的抑制作用(P<0.05)。结论：沉默HNF1A-AS1可通过靶向上调miR-1表达减弱膀胱癌细胞的增殖和侵袭能力。

Effects and mechanisms of HNF1A-AS1 on cell proliferation and invasion of bladder cancer cells via targeting miR-1

Objective:To investigate the effects of HNF1A-AS1 on bladder cancer cells via target miR-1.Methods: The miR-1 level of bladder cancer cell lines and normal urinary bladder epithelial cell line was measured by RT-PCR.After silencing or upregulating the expressions of HNF1A-AS1 and miR-1,the level of miR-1 was detected again.Luciferase reporter assay was performed to determine the relationship of HNF1A-AS1 and miR-1.And the cell proliferation,invasion and related protein levels were detected by MTT,Transwell and Western blot assay respectively.Results: The 253J cell line was selected for further study according to the results of pre-experiment.Overexpression of HNF1A-AS1 inhibited the expression of miR-1 significantly and alleviated the promotive effect of miR-1 mimic on miR-1(P<0.05).And downregulated HNF1A-AS1 induced the expression of miR-1 and attenuated the inhibitory effect of miR-1 inhibitor makedly(P<0.05).Meanwhile,the targeted relationship of HNF1A-AS1 and miR-1 was demonstrated by luciferase reporter assay.In addition,silencing of HNF1A-AS1 inhibited proliferation and invasion coupled with decreased Ki67,PCNA,MMP-9 and VEGF(P<0.05).Furthermore,knockdown of miR-1 reversed the inhibitory effects of HNF1A-AS1 on proliferation and invasion(P<0.05).Conclusion: Downregulation of HNF1A-AS1 attenuates the cell proliferation and migration abilities of bladder cancer cells through inhibiting miR-1.