›› 2017, Vol. 37 ›› Issue (8): 1051-.doi: 10.3969/j.issn.1674-8115.2017.08.001

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Mechanism of hsa-miRNA124-3p regulating the proliferation and migration of human lung cancer cell line NCI-H460#br#

WANG Ye, XIE Wang, ZHANG Jie, SHENG Jie-ying, GUO Zhong-liang   

  1. Department of Respiration Medicine, Shanghai East Hospital, Tongji University, Shanghai 200120, China
  • Online:2017-08-28 Published:2017-09-28
  • Supported by:
     National Natural Science Foundation of China,81370174)

Abstract: Objective · To study the regulation of hsa-miRNA124-3p on the proliferation and migration of human lung cancer NCI-H460 cells and its mechanism.  Methods · Four pairs of lung cancer and para-carcinoma tissues were harvested in clinical and measured for hsa-miRNA124-3p and Krüppellike factor 4 (KLF4) levels. The theoretical binding site of hsa-miRNA124-3p in 3’-UTR of KLF4 was predicted by bioinformatics, and validated by luciferase report assay. NCI-H460 cells were transfected with pshRNA-Sponge-miRNA124 or pshRNA-KLF4, and 48 hours later, the proliferation of NCI-H460 cells after genetic intervention was assayed by the MTT method, and cell migration ability was observed by streak method.  Results · For all four pairs of samples tested, hsa-miRNA124-3p was higher in the cancer tissues than in the adjacent tissue (P<0.01), and KLF4 protein was lower in the cancer tissues than in the adjacent tissue (P<0.01). The bioinformatic analysis showed there is a theoretical binding site (5’-UGCCUUAA-3’) of hsa-miRNA124-3p in 3’-UTR of KLF4. Luciferase activity assay showed that hsa-miRNA124-3p could bind to the 3’-UTR region of KLF4 gene and negatively regulate the expression of protein. The proliferation of NC-H460 cells was suppressed by transfection with pshRNA-Sponge-miRNA124 72 h after transfection (P<0.05 ). Compared with the control group, the proliferation activity of pshRNA-KLF4 transfection group was further enhanced (P<0.05) There was no significant difference in the proliferation of pshRNA-Sponge-miRNA124 and pshRNA-KLF4 cotransfection group and the control group (P>0.05). The data of cell migration assay showed that the changes of cell migration ability were the same as proliferation activity of the cells in groups 72 h after transfection.  Conclusion · Hsa-miRNA124-3p increases the proliferation and migration in NCI-H460 cells via suppressing the expression of KLF4, and reducing the content of miRNA124-3p in NC-H460 cells can inhibit cell proliferation and migration via upregulating KLF4 expression.

Key words:  lung cancer, Krüppel-like factor 4, NCI-H460, hsa-miRNA124-3p, proliferation, migration