Volume 43 Issue 12
Dec.  2022
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Wei ZHANG, Baoquan WANG, Xifeng LEI, Xu WANG, Liang ZHANG. miR-125b-5p Regulates HK2 to Inhibit Proliferation and Glycolysis of Gallbladder Cancer Cells[J]. Journal of Kunming Medical University, 2022, 43(12): 23-29. doi: 10.12259/j.issn.2095-610X.S20221206
Citation: Wei ZHANG, Baoquan WANG, Xifeng LEI, Xu WANG, Liang ZHANG. miR-125b-5p Regulates HK2 to Inhibit Proliferation and Glycolysis of Gallbladder Cancer Cells[J]. Journal of Kunming Medical University, 2022, 43(12): 23-29. doi: 10.12259/j.issn.2095-610X.S20221206

miR-125b-5p Regulates HK2 to Inhibit Proliferation and Glycolysis of Gallbladder Cancer Cells

doi: 10.12259/j.issn.2095-610X.S20221206
  • Received Date: 2022-09-15
    Available Online: 2022-12-05
  • Publish Date: 2022-12-25
  •   Objective  To investigate the mechanism of miR-125b-5p regulating the proliferation and glycolysis of gallbladder cancer cells by HK2.   Methods  NC mimic, miR-125b-5p mimic, miR-125b-5p mimic+pcDNA-NC and miR-125b-5p mimic+pcDNA-HK2 were transfected into human gallbladder carcinoma GBC-SD cells, respectively. Real Time Quantitative Polymerase Chain Reaction (RT-qPCR) was used to detect the expression of miR-125b-5p and HK2 mRNA. The protein expressions of HK2, LDHA and PDK1 were detected by Western blot. Cell counting Kit-8 (CCK-8) was used to measure cell proliferation activity, and the glycolysis kit was used to measure lactic acid production, glucose consumption, and ATP production. Dual luciferase reporter assay was used to verify the targeting relationship between miR-125b-5p and HK2.   Results  The expression of miR-125b-5p was reduced in gallbladder carcinoma cell lines GBC-SD, NOZ (P < 0.0001) and tissues (P < 0.01), and was lower in GBC-SD cells than in NOZ cells. Overexpression of miR-125b-5p significantly inhibited the proliferation of GBC-SD cells (P < 0.05), glucose intake (P < 0.05), the production of lactic acid (P < 0.001) and ATP production (P < 0.01), and the expression of LDHA (P < 0.01), PDK1 (P < 0.001), and HK2 (P < 0.0001). starBase database and dual luciferase experiments confirmed that miR-125b-5p targeted HK2 and negatively regulated its expression. Cell proliferation rate (P < 0.01), glucose consumption (P < 0.05), lactic acid (P < 0.01) and ATP production (P < 0.05), and the expression of LDHA (P < 0.05), PDK1 ( P < 0.05)、HK2 (P < 0.05)were significantly higher in the miR-125b-5p and HK2 overexpression groups.   Conclusion  Overexpression of miR-125b-5p inhibited the proliferation and aerobic glycolysis in human gallbladder carcinoma cell GBC-SD through targeting HK2.
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