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Archive > Volume 30 Issue 12 > 2023,30(12):1066-1073. DOI:10.3872/j.issn.1007-385X.2023.12.005 Prev Next
Basic Research
miR-4465 inhibits the malignant biological behaviors of hepatocellular carcinoma Hep3B cells by targeting and downregulating the expression of HMGA1
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Abstract:
Objective: To investigate the effect of miR-4465 targeting high mobility group protein A1 (HMGA1) on the proliferation, migration and invasion of hepatocellular carcinoma Hep3B cells. Methods: Sixteen pairs of cancer tissues and adjacent tissue samples from patients diagnosed with hepatocellular carcinoma in the First Affiliated Hospital of Wannan Medical College from May 2020 to September 2021 were collected. The expression of miR-4465 in hepatocellular carcinoma tissues and Hep3B and Huh7 cells was analyzed by qPCR, and the regulatory relationship between miR-4465 and HMGA1 was verified by dual luciferase reporter gene assay. According to the different transfections, Hep3B cells were group, si-NC group, and si-HMGA1 group for transfection; in addition, mimics-NC+pcDNA-NC, miR-4465 mimics+pcDNA-NC, and miR-4465 mimics+pcDNA-HMGA1 were transfected for rescue experiments. The changes of mRNA and protein levels of HMGA1 in each group were detected by qPCR and WB assay; the changes in cell proliferation activity in each group were detected by CCK-8 method; the changes in cell migration ability in each group were detected by scratch assay, and the changes in cell invasion ability in each group were detected by Transwell assay. Results: The expression of miR-4465 in hepatocellular carcinoma tissues and cells was significantly lower than that in adjacent tissues and normal liver cells (P<0.05 or P<0.001). After transfection for 48 h, the proliferation, migration and invasion abilities of Hep3B cells overexpressing miR-4465 were significantly decreased (P<0.05, P<0.01 or P<0.001); the proliferation, migration and invasion abilities of Hep3B cells with miR-4465 knockdown were increased (P<0.05, P<0.01 or P<0.001). The binding relationship between HMGA1-3'UTR and miR-4465 was verified by dual luciferase report gene experiment. miR-4465 could target and down-regulate the mRNA and protein expression of HMGA1 (all P<0.01). Over-expression of HMGA1 partially restored the inhibitory effects of miR-4465 overexpression on cell proliferation, migration and invasion as well as HMGA1 expression (all P<0.05). Conclusion: miR-4465 inhibits the malignant biological behaviors of hepatocellular carcinoma Hep3B cells by down-regulating the expression of HMGA1 in the cells.
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