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Highly expressed miRNA let-7i-5p in renal cell carcinoma tissues regulates the malignant biological behaviors of 769-P cells through hyaluronan-binding protein 4
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Abstract:
Objective: To investigate the expression level of miRNA let-7i-5p in renal cell carcinoma (RCC) tissues and to explore its effect on the proliferation, migration, invasion, and hyaluronan-binding protein 4 (HABP4) expression in human RCC 769-P cells.Methods: A meta-analysis of let-7i-5p expression in RCC tissues was performed using the TCGA RCC database and GEO database.The human RCC 769-P cells were routinely cultured for transfection in vitro, and were divided into three groups according to different transfection materials: overexpression group (transfected with let-7i-5p mimics), inhibition group (transfected with let-7i-5p inhibitors),and control group (transfected with NC sequence). CCK-8, cell scratch test, Transwell assay, and WB method were used to detect the effects of let-7i-5p on the proliferation, scratch healing rate, invaded cell numbers and HABP4 expression in 769-P cells. Dual[1]luciferase reporter gene assay was performed to demonstrate the targeting relationship between let-7i-5p and HABP4. Results: Data analysis of TCGA RCC database and 5 GEO data sets (GSE23085, GSE47582, GSE95385, GSE16441, and GSE71302) showed that the expression level of let-7i-5p was significantly higher in RCC tissues than in normal kidney tissues (all P <0.05). As shown by in vitro experiments, compared with the control group, the cell proliferation activity of the overexpression group was significantly increased at 24, 48, and 72 h, while that of the inhibition group was decreased (all P<0.01). The scratch healing rate [(37.276±2.058)% vs (15.663±2.949)%, P<0.01] and the invaded cell numbers [(377.000±34.044) vs (255.667±25.34), P<0.05] were significantly increased in the overexpression group, while the scratch healing rate [(8.791±2.568) % vs (15.663±2.949) %, P<0.05] and the invaded cell numbers [(170.333±14.978) vs (255.667±25.384), P<0.01] were significantly reduced in the inhibition group. The luciferase activity was significantly inhibited by the overexpression of let-7i-5p in the wild-type HABP4-3'URT plasmid group (P<0.01), while no statistical difference was observed in the mutant HABP4-3'URT plasmid group (P>0.05). WB results showed that compared with the control group, the expression of HABP4 and E-cadherin in the overexpression group was decreased (all P<0.01), and the expression of CDK2 was increased (P<0.01), while the inhibition group showed the opposite trend (all P<0.01). Conclusion: Let-7i-5p is highly expressed in RCC tissues and promotes the proliferation, migration, and invasion of 769-P cells possibly by targeting HABP4.
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Project Supported:
Project supported by the Youth Foundation of Southwest Medical University (No. 2018-ZRQN-142; No. 2018-ZRQN-041)
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