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miRNA-26b-3p regulates proliferation and migration of esophageal squamous cell carcinoma cells by targeting STAT3
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Abstract:
Objective: To detect the expression level of miR-26b-3p in esophageal squamous cell carcinoma (ESCC) tissues, and to explore its effect on the proliferation, invasion and migration of ESCC cells and its molecular mechanism. Methods: 60 pairs of ESCC tissues and corresponding adjacent tissues that surgically resected from April 1, 2018 to December 25, 2018 were collected from the Fourth Hospital of Hebei Medical University. qPCR was used to detect the expression of miR-26b-3p in ESCC tissues, corresponding adjacent tissues and ESCC cells. ESCC cell lines TE1 and KYSE150 with low miR-26b-3p expression were selected and transfected with miR-26b-3p mimics. At the same time, the negative control was set up. The effects of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells were detected by cell proliferation assay, Wound healing assay and Transwell assay. Luciferase reporter gene analysis was used to detect the binding between miR-26b-3p and STAT3 3'UTR. Then, miR-26b-3p mimics and pcDNA3.0-STAT3 were co-transfected into the cells to verify whether STAT3 could reverse the effect of miR-26b-3p on cell proliferation, migration and invasion using cell proliferation assay, Wound healing assay and Transwell test. The effects of methylase inhibitor 5-Aza-DC on the methylation of ESCC cells as well as the expression of miR-26b-3p and STAT3 were detected by qPCR and WB. Results: The expression of miR-26b-3p in ESCC tissues was lower than that in the adjacent normal tissues (P<0.05), and the expression was also significantly lower in ESCC cell lines (TE1, KYSE150 and LYSE170) as compared with normal human esophageal epithelial HEEC cells (all P<0.01). Compared with miR-26b-3p NC group, transfection of miR-26b-3p mimics significantly up[1]regulated the expression of miR-26b-3p in TE1 and KYSE150 cells and obviously inhibited the proliferation, invasion and migration of the cells (P<0.05 or P<0.01). The luciferase reporter gene assay showed that miR-26b-3p significantly inhibited the luciferase activity of wild-type STAT3 vector in TE1 and KYSE150 cells (P<0.05 or P<0.01), while the luciferase activity of mutant-type was not affected. Co-transfection with miR-26b-3p mimics and pcDNA3.0-STAT3 partially reversed the inhibitory effect of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells (P<0.05 or P<0.01). After 5-aza-DC treatment, the expression of miR[1]26b-3p was up-regulated (P<0.01), mRNA and protein levels of STAT3 were decreased (P<0.05), and miR-26b-3p was demethylated in TE1 and KYSE150 cells. Conclusion: miR-26b-3p is down-regulated in ESCC tissues and cells due to the hypermethylation of its promoter. As a tumor suppressor, miR-26b-3p inhibits the proliferation, invasion and migration of ESCC cells via targeting STAT3.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81972722), and the Youth Science and Technology Project from Health Commission of Hebei Prorince (No. 20190720)
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