lncRNANORADpromotestheproliferationandmigrationofesophagealsquamous cell carcinoma EC9706 cells / 中国肿瘤生物治疗杂志

ABSTRACT

@#[Abstract] Objective: To investigate the effects and mechanisms of long non-coding RNA (lncRNA) non-coding RNA-activated by DNA damage (NORAD) on the proliferation and migration of esophageal squamous cell carcinoma (ESCC) EC9706 cells. Methods: RT-PCR was used to detect the mRNA expression level of NORAD in different ESCC cells (EC9706, TE1, YES-2, KYSE150). Small interfering RNA (siRNA) targeting NORAD gene was transfected into EC9706 cells (as si-NORAD group) with RNA interference technique to knockdown NORAD expression; in addition, blank control group (as Ctrl group, without any transfection) as well as negative control group (as NC group, transfected with siRNAnegative control sequence)werealsoestablished. qPCR was used to verify the transfection efficiency. MTT, Colony formation assay and Wound-healing test were used to detect the abilities of proliferation and migration of EC9706 cells before and after NORAD knockdown. Western blotting was used to detect the expressions of E-cadherin, N-cadherin and Snail in EC9706 cells before and after NORAD knockdown. Results: NORAD mRNAwas highly expressed in 4 ESCC cell lines. Comparing with TE1, YES-2 and KYSE150 cells, the expression of NORAD mRNA was significantly higher in EC9706 cells (P<0.01). After transfection of NORAD-siRNA into EC9706 cells, the expression of NORAD was down-regulated significantly as comparing with Ctrl group and NC group (all P<0.01), in the meanwhile, the proliferation and migration abilities of EC9706 cells were also significantly suppressed (P<0.05).After NORAD knockdown, the expression of E-cadherin was up-regulated while the expressions of N-cadherin and Snail were down-regulated in EC9706 cells (all P<0.05). Conclusion: NORAD is highly expressed in EC9706 cells;knockdown of NORAD expression can inhibit the proliferation and migration ability of EC9706 probably through up-regulating E-cadherin and down-regulating N-cadherin and Snail.