Effects of IKCa1 channels on migration,invasion,and miRNA expression profile of human cervical cancer HeLa cells

CHENG hui; GAO Lanyang; ZHAN Ping; MAO Xiguang

doi:10.3969/j.issn.1002-266X.2017.41.007

Abstract：Objective To investigate the effects of IKCa1 channels on the migration,invasion,and miRNA expres-sion profile of cervical cancer HeLa cells. Methods HeLa cells in the logarithmic phase were randomly divided into the observation groups 1-5 and the DMSO group,which were then treated with 0,10. 0,20. 0,30. 0,and 40. 0 umol/ L TRAM-34 (a characteristic inhibitor of IKCa1 channel),and equal volume of DMSO (TRAM-34 solvent),respectively, for 24,48 and 72 h. The migration characteristics of HeLa cells were detected by Scratch test. Transwell invasion assay was used to detect invasive properties. The miRNA expression profile of IKCa1 channel was detected by high-throughput se-quencing,and some differentially expressed miRNAs were verified by qRT-PCR. The miRanda software was used to predict the target genes that could differentiate miRNAs and underwent signal pathway enrichment analysis. Results Scratch test suggested that TRAM-34 inhibited the migration of HeLa cells,with a concentration- and time-dependent manner (P <0. 001). Transwell invasion showed that the invasion rate of HeLa cells decreased with the increase of TRAM-34 concentra-tions (F = 384. 050,P < 0. 001). High-throughput sequencing revealed 15 differentially expressed and up-regulated miR-NAs and 5 down-regulated miRNAs. The results of qRT-PCR showed that the differential expression of miRNAs was con-sistent with the sequencing results. Bioinformatics analysis showed that the target gene of hsa-miR-143-5p aggregated in the cancer-related pathways,p53 gene pathway,and MAPK signal pathway,while the target gene of hsa-miR-421 aggregated in the tumor proteoglycans,Rap1 signal pathway,and Wnt signaling pathway. Conclusion Blocking IKCa1 channel can in-hibit the migration and invasion of HeLa cells and influence the miRNAs expression profile.