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Archive > Volume 28 Issue 5 > 2021,28(5):489-494. DOI:10.3872/j.issn.1007-385X.2021.05.010 Prev Next
Clinical Research
Effect of miRNA-490-3p regulating Smad2/TGF- β on sensitivity of colorectal cancer SW480 cells to oxaliplatin
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Abstract:
[Abstract] Objective: To investigate the effect of miRNA-490-3p regulating Smad2/TGF-β on chemo-sensitivity of colorectal cancer to oxaliplatin. Methods: One hundred patients with colorectal cancer (CRC) who received oxaliplatin (OXA) chemotherapy after radical operation were selected as research subjects and divided into drug resistant group (n=40) and non-resistant group (n=60) according to the chemotherapy outcome. The level of miRNA-490-3p in peripheral blood of two groups of patients was detected by qPCR. Human CRC cell line SW480 and oxaliplatin resistant CRC cell line OXA-SW480 were selected for the study. The expression level of miRNA-490-3p in the two cell lines was detected by qPCR, and then the miRNA-490-3p over-expression vector was transfected into OXA-SW480 cells (over-expression group), in the meanwhile, the empty vector group (OXA-SW480 cells transfected with empty vector) and blank control group (OXA-SW480 cells without any treatment) were set up. CCK-8 was used to detect cell proliferation ability of each group, and different concentrations of OXA were used to treat cells of each group to calculate the half inhibitory concentration (IC50); Annexin Ⅴ-FITC/PI staining was used to detect cell apoptosis rate of each group; WB assay was used to detect the expression levels of Smad2 and TGF-β. Results: In CRC patients, the level of miR-490-3p in the peripheral blood of the drug-resistant group was significantly lower than that of the non-resistant group. In the CRC cells, the level of miR-490-3p in the OXA-SW480 cells was significantly lower than that of the normal SW480 cells (P<0.05). Compared with the empty vector group and the blank control group, the level of miR-490-3p in the over-expression group was significantly increased (all P<0.05), the proliferation inhibition rate and the apoptosis rate were significantly increased (all P<0.05), Smad2 protein level was significantly reduced (all P<0.05), and TGF-β protein level was significantly increased (all P<0.05). After OXA treatment, the IC50 of the over-expression group was significantly lower than that of the empty vector group and the blank control group (both P<0.05). According to the results of Pearson correlation analysis, the expression of miR-490-3p was negatively correlated with the expression of Smad2 (r=–0.943, P<0.01), but positively correlated with the expression of TGF-β (r=0.961, P<0.01). Conclusion: Over-expression of miRNA-490-3p can increase the OXA sensitivity of CRC SW480 cells, which is related to the Smad2/TGF-β signaling pathway.
Keywords:
Project Supported:
Project supported by the Medical Science and Technology Research Fund Project of Guangdong Province (No. A2016500), and the Self-Financing Science and Technology Project of Guangdong Province（No.2017ZC0250）
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