lncRNA SNHG1靶向miR-7对结直肠癌细胞放疗抵抗的影响

Influence of lncRNA SNHG1 on radioresistance of colorectal cancer cells by targeting miR-7

目的:探讨lncRNA SNHG1通过miR-7对结直肠癌(CRC)细胞放疗抵抗的影响。方法:将CRC细胞HT29分为ctrl组、si-NC组、si-SNHG1组、si-SNHG1+inhibitor-NC组、si-SNHG1+miR-7inhibitor组,qRT-PCR检测细胞中SNHG1、miR-7的表达;克隆形成实验检测细胞放射敏感性;MTT法检测细胞增殖;流式细胞仪检测细胞凋亡;Western blot检测细胞中PCNA、Bax、Caspase-3、GLUT1、HK2的表达;双荧光素酶报告基因实验验证SNHG1与miR-7的关系。结果:与ctrl组、si-NC组比较,si-SNHG1组的HT29细胞的PE、SF、A490值、PCNA、GLUT1、HK2表达明显降低(P<0.05),细胞凋亡率、Bax、Caspase-3表达明显上升(P<0.05);与si-SNHG1组、si-SNHG1+inhibitor-NC组比较,si-SNHG1+miR-7 inhibitor组HT29细胞的PE、SF、A490值、PCNA、GLUT1、HK2表达显著升高(P<0.05),细胞凋亡率、Bax、Caspase-3表达明显降低(P<0.05);双荧光素酶报告显示,SNHG1靶向调控miR-7表达。结论:敲低SNHG1可能通过靶向调控miR-7,进而改善CRC的放疗抵抗。

Objective:To investigate the influence of long non-coding RNA small nucleolar RNA host gene 1(lncRNA SNHG1)on the radioresistance of colorectal cancer(CRC)cells via miR-7.Methods:CRC cells HT29 were separated into:ctrl group(normally cultured HT29 cells),si-NC group(transfected with si-NC),si-SNHG1 group(transfected with si-SNHG1),si-SNHG1+inhibitor-NC Group(co-transfected with si-SNHG1 and inhibitor-NC),and si-SNHG1+miR-7 inhibitor group(co-transfected with si-SNHG1 and miR-7 inhibitor);qRT-PCR was applied to detect the expression of SNHG1 and miR-7 in cells;clonogenic assays were applied to detect cellular radiosensitivity;MTT assay was applied to detect cell proliferation;flow cytometry was applied to detect apoptosis;Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),cysteine protease 3(Caspase-3),glucose transporter 1(GLUT1),hexokinase 2(HK2);and dual-luciferase reporter assays were applied to verify the relationship between SNHG1 and miR-7.Results:Compared with ctrl group and si-NC group,the colony formation rate(PE),survival fraction(SF),A490 value,PCNA,GLUT1,HK2 expression of HT29 cells in si-SNHG1 group were obviously decreased(P<0.05),the apoptosis rate,and the Bax,Caspase-3 expression were obviously increased(P<0.05);compared with si-SNHG1 group and si-SNHG1+inhibitor-NC group,the PE,SF,A490 value,PCNA,GLUT1,HK2 expression of HT29 cells in si-SNHG1+miR-7 inhibitor group were obviously increased(P<0.05),the apoptosis rate,and the Bax and Caspase-3 expression were obviously decreased(P<0.05);dual-luciferase reporter revealed that SNHG1 targeted miR-7 expression.Conclusion:Knockdown of SNHG1 may improve the radioresistance of CRC by targeting and regulating miR-7.