长链非编码RNA ABHD11-AS1对结直肠癌细胞生长和转移的影响及调控机制

Effect of long noncoding RNA ABHD11-AS1 on colorectal cancer cell growth and metastasis and the regulation mechanism

目的 探讨长链非编码RNA(lncRNA)α/β水解酶域11反义RNA1(ABHD11-AS1)对结直肠癌细胞生长和转移的影响,并基于miR-133a/真核起始因子4A1(EIF4A1)轴分析其可能的调控机制。方法 (1)取人结直肠癌细胞HT-29、SW480、DLD-1、HCT116、LOVO、SW620,以及正常结直肠黏膜细胞FHC,检测细胞中lncRNA ABHD11-AS1的表达情况。(2)分别通过miRcode、TargetScan数据库预测ABHD11-AS1与miR-133a、miR-133a与EIF4A1的靶向关系,并进行双荧光素酶报告基因实验以验证。(3)将HT-29细胞分为NC组(转染Negative Control)、siABHD11-AS1组(转染lncRNA ABHD11-AS1 siRNA)、miR-133a inhibitor组(转染miR-133a inhibitor)、siABHD11-AS1+miR-133a inhibitor组(转染lncRNA ABHD11-AS1 siRNA和miR-133a inhibitor)和siABHD11-AS1+miR-133a inhibitor+siEIF4a1组(转染lncRNA ABHD11-AS1 siRNA、miR-133a inhibitor、EIF4A1 siRNA)。检测转染后除miR-133a inhibitor组以外的其他组细胞增殖、迁移和侵袭能力,NC组、siABHD11-AS1组、miR-133a inhibitor组、siABHD11-AS1+miR-133a inhibitor组细胞的EIF4A1蛋白表达量。结果 (1)ABHD11-AS1均高表达于各种结直肠癌细胞,在HT-29细胞中的表达最高。(2)miR-133a为ABHD11-AS1靶基因,EIF4A1为miR-133a靶基因。(3)siABHD11-AS1组HT-29细胞的增殖、迁移和侵袭能力均低于NC组(均P<0.05),siABHD11-AS1+miR-133a inhibitor组HT-29细胞的增殖、迁移和侵袭能力均高于siABHD11-AS1组(均P<0.05),siABHD11-AS1+miR-133a inhibitor+siEIF4A1组HT-29细胞的增殖、迁移和侵袭能力均低于siABHD11-AS1+miR-133a inhibitor组(均P<0.05)。(4)miR-133a inhibitor组HT-29细胞中EIF4A1蛋白表达量高于NC组(P<0.05);siABHD11-AS1组HT-29细胞中EIF4A1蛋白表达量低于NC组,而siABHD11-AS1+miR-133a inhibitor组HT-29细胞中EIF4A1蛋白表达量高于siABHD11-AS1组(均P<0.05)。结论 ABHD11-AS1在结直肠癌细胞中呈高表达,抑制其表达后结直肠癌细胞的生长和转移能力均下降,其可能通过调控miR-133a/EIF4A1信号轴发挥作用。

Objective To investigate the effect of long noncoding RNA(lncRNA)alpha/beta hydrolase domain-containing protein 11 antisense RNA 1(ABHD11-AS1)on the growth and metastasis of colorectal cancer cells,and to analyze its possibly regulation mechanism based on miR-133 a/eukaryotic initiation factor 4 A1(EIF4 A1)axis.Methods(1)The human colorectal cancer cells HT-29,SW480,DLD-1,HCT116,LOVO,SW620,and normal colorectal mucosal cells FHC were obtained,and the expression of lncRNA ABHD11-AS1 in the cells was detected.(2)By databases of miRcode and TargetScan,the targeted relations of ABHD11-AS1 with miR-133 a,and of miR-133 a with EIF4 A1 were predicted;moreover,the dual luciferase reporter gene assay was employed to verify.(3)HT-29 cells were assigned to the NC group(transfected with Negative Control),the siABHD11-AS1 group(transfected with lncRNA ABHD11-AS1 siRNA),the miR-133 a inhibitor group(transfected with miR-133 a inhibitor),the siABHD11-AS1+miR-133 a inhibitor group(transfected with lncRNA ABHD11-AS1 siRNA and miR-133 a inhibitor),or the siABHD11-AS1+miR-133 a inhibitor+siEIF4 A1 group(transfected with lncRNA ABHD11-AS1 siRNA,miR-133 a inhibitor,and EIF4 A1 siRNA).After transfection,except for the miR-133 a inhibitor group,the cell proliferation,metastasis and invasion in the remaining groups,and the EIF4 A1 protein expression of cells in the NC,siABHD11-AS1,miR-133 a inhibitor,and siABHD11-AS1+miR-133 a inhibitor groups were detected.Results(1)ABHD11-AS1 was highly expressed in various colorectal cancer cells,and exhibited the highest expression in HT-29 cells.(2)miR-133 a was the target gene of ABHD11-AS1,and EIF4 A1 was the target gene of miR-133 a.(3)The siABHD11-AS1 group depicted lower proliferation,metastasis and invasion of HT-29 cells as compared with the NC group(all P<0.05),whereas the siABHD11-AS1+miR-133 a inhibitor group yielded higher proliferation,metastasis and invasion of HT-29 cells in the comparison of the siABHD11-AS1 group(all P<0.05).The siABHD11-AS1+miR-133 a inhibitor+siEIF4 A1 group exhibited lower proliferation,metastasis and invasion of HT-29 cells compared to the siABHD11-AS1+miR-133 a inhibitor group(all P<0.05).(4)The miR-133 a inhibitor group interpreted a higher EIF4 A1 protein expression of HT-29 cells as compared with the NC group(P<0.05);the siABHD11-AS1 group had a lower EIF4 A1 protein expression of HT-29 cells in the comparison of the NC group,whereas the siABHD11-AS1+miR-133 a inhibitor group implied a higher EIF4 A1 protein expression of HT-29 cells compared to the siABHD11-AS1 group(all P<0.05).Conclusion ABHD11-AS1 is highly expressed in colorectal cancer cells,and the growth and metastasis of colorectal cancer cells decline after the inhibition of ABHD11-AS1 expressions,possibly exerting the effects by regulating miR-133 a/EIF4 A1 signaling axis.