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| Effect of long non-coding RNA HOX transcript antisense inter genic RNA targeting miR-206 on migration and invasion of colorectal cancer cells |
| XUE Qiang1 WU Xueliang2 SUN Guangyuan3 YANG Dongdong3 LI Kun4 |
1.Department of General Surgery, the First Hospital of Zhangjiakou, Hebei Province, Zhangjiakou 075000, China;
2.Cancer Research Institute, the First Affiliated Hospital of Hebei North University, Hebei Province, Zhangjiakou 075000, China;
3.Department of General Surgery, the First Affiliated Hospital of Hebei North University, Hebei Province, Zhangjiakou 075000, China;
4.Department of Pathology, the First Affiliated Hospital of Hebei North University, Hebei Province, Zhangjiakou 075000, China |
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Abstract Objective To investigate the effect of long noncoding RNA (lncRNA) HOX transcript antisense inter genic RNA (HOTAIR) targeting miR-206 on invasion and migration of colorectal cancer cells. Methods HT-29 cells of human colorectal cancer were cultured regularly, and the synthetic HOTAIR siRNA and siRNA control were transfected into HT-29 cells as si-HOTAIR group and si-con group, and the non-transfected HT-29 cells were set as blank group. Real time-quantitative polymerase chain reaction was used to determine the transfection effect, and Transwell and scratch healing tests were used to detect the invasion and migration of HT-29 cells. Bioinformatics software analyzed the binding sites of HOTAIR and miR-206 region, and constructed mutant (HOTAIR 3’UTR mut) and wild type (HOTAIR 3’UTR wt) luciferase reporter vectors. HOTAIR 3’UTR wt and HOTAIR 3’UTR mut were transfected into HT-29 cells with miR-206 mimic or miR-NC vector. HOTAIR 3’UTR wt/miR-NC group, HOTAIR 3’UTR wt/miR-206 group, HOTAIR 3’UTR mut/miR-NC group and HOTAIR 3’UTR mut/miR-206 group were cultured for 48 h to detect luciferase activity. HOTAIR siRNA and miR-206 inhibitor were co-transfected into HT-29 cells, si-HOTAIR+anti-NC group and si-HOTAIR+anti-miR-206 group were set up, and the cell invasion and migration levels were analyzed by Transwell and scratch healing assay. Results There was no significant difference in the expression level of HOTAIR mRNA in HT-29 cells between blank group and si-con group (P＞0.05). Compared with si-con group, the expression level of HOTAIR mRNA in HT-29 cells in si-HOTAIR group was decreased (P＜0.01). There was no significant difference in HT-29 cell invasion number and cell scratch healing rate between blank group and si-con group (P＞0.05). Compared with the si-con group, the invasion number of HT-29 cells in the si-HOTAIR group was decreased, and the cell scratch healing rate was decreased (P＜0.01). Compared with HOTAIR 3’UTR wt/miR-NC group, the luciferase activity of HOTAIR wt in the HOTAIR 3’UTR wt/ miR-206 group decreased (P＜0.01). There was no significant difference in fluorescein activity between HOTAIR 3’UTR mut/miR-NC group and HOTAIR 3’UTR mut/miR-206 group (P＞0.05). Compared with si-HOTAIR+anti-NC group, the expression level of miR-206 in si-HOTAIR+anti-miR-206 group was decreased (P＜0.01). Compared with si-HOTAIR+anti-NC group, the invasion and mobility of HT-29 cells in si-HOTAIR+anti-miR-206 group were increased (P＜0.01). Conclusion HOTAIR promotes the invasion and migration of HT-29 cells by negatively regulating miR-206.
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