Effect of microRNA-212-5p on the biology of colorectal cancer cells by regulating ATP-binding box E1
Objective To explore the role and mechanism of microRNA(miR)-212-5p in the pro-gression of colorectal cancer(CRC).Methods CRC tissues and adjacent normal tissues of 32 patients were collected from the First Affiliated Hospital of Xinxiang Medical College,and normal colon mucosal ep-ithelial cells(NCM460)and CRC cell lines(SW480,SW1116,HT29)purchased from the Cell Bank of Shanghai Chinese Academy of Sciences were cultured in vitro.The expression of miR-212-5p and ATP-binding box E1(ABCE1)was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).SW480 cells were divided into miR-NC group,miR-212-5p group,si-NC group,si-ABCE1 group,miR-212-5p+pcDNA group,miR-212-5p+pcDNA-ABCE1 groups.Methyl thia-zolyl tetrazolium(MTT)method and Transwell analysis were used to detect the proliferation,migration and invasion ability of SW480 cells in vitro.Western blotting was used to detect the ABCE1 protein expression.The targeting relationship between miR-212-5p and ABCE1 was detected by double luciferase assay.The difference between two groups of data was tested by independent sample t test,and the difference between multiple groups of data was tested by one-way ANOVA and SNK-q test.Results MiR-212-5p expression in CRC tissue and cells was significantly decreased(tissue:t=100.411,P＜0.01;cell:F=1 192.235,P＜0.01),while ABCE1 mRNA(tissue:t=85.952,P＜0.01;cell:F=219.953,P＜0.01)and pro-tein(tissue:t=39.634,P＜0.01;cell:F=125.185,P＜0.01)expression was significantly increased.Compared with the miR-NC group,the cell viability(48 h:0.34±0.01 vs.0.53±0.03,t=18.025,P＜0.01;72 h:0.51±0.02 vs.0.97±0.08,t=16.735,P＜0.01),the number of migrating cells(40.31±2.12 vs.87.42±6.85,t=19.710,P＜0.01),the number of invasive cells(29.06±1.35 vs.72.19±5.91,t=21.344,P＜0.01)and the expression of ABCE1 protein(0.17±0.02 vs.0.56± 0.05,t=21.726,P＜0.01)of SW480 cells in the miR-212-5p group were significantly reduced.Com-pared with the si-NC group,the cell viability(48 h:t=10.436,P＜0.01;72 h:t=17.332,P＜0.01),the number of migrating cells(t=16.741,P＜0.01)and the number of invasive cells(t=17.164,P＜0.01)of SW480 cells in the si-ABCE1 group were significantly reduced.Compared with the miR-212-5p+pcDNA group,the cell viability(48 h:q=21.689,P＜0.01;72 h:q=16.709,P＜0.01),the number of migrating cells(q=20.273,P＜0.01),the number of invasive cells(q=29.325,P＜0.01)and the expression of ABCE1 protein(q=24.000,P＜0.01)of SW480 cells in the miR-212-5p+pcDNA-ABCE1 group were significantly increased.Conclusion MiR-212-5p inhibits the biological behavior of CRC cells by targeting ABCE1.
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