Effects of Butorphanol on the Proliferation and Apoptosis of Gastric Cancer Cells by Regulating miR-212-3p/KDM5B Axis
ZHONG Weirong*, ZHOU Yang
This study aims to discuss the impacts of butorphanol on the proliferation and apoptosis of gas tric cancer cells by regulating miR-212-3p (microRNA-212-3p)/KDM5B (lysine-specific demethylase 5B) axis. MKN-45 cells were grouped into Control group, butorphanol low concentration group (B-L, 1 μmol/L), medium concentration group (B-M, 2 μmol/L), and high concentration group (B-H, 4 μmol/L), B-H+anti-miR-NC group, B-H+anti-miR-212-3p group, miR-NC group, and miR-212-3p group. RT-qPCR was performed to measure miR-212-3p, KDM5B mRNA. Dual luciferase reporter assay was performed to measure the targeting relationship between miR-212-3p and KDM5B. Cell proliferation was detected by Edu method; cell invasion was detected Trawesll method; the scratch healing test was used to detect cell migration. Cell apoptosis was detected by flow cytometry. Western blot was performed to detect the protein levels of Ki-67, MMP-2, MMP-9, Bax, Bcl-2, KDM5B, Caspase 3, Caspase 7, Cleaved Caspase 3 and Cleaved Caspase 7. Dual luciferase assay indicated that miR-212-3p had a target relationship with KDM5B. Compared with the Control group, number of Edu positive cells, number of inva sion cells, and scratch healing rate of MKN-45 cells in the B-L, B-M, B-H groups decreased, the protein levels of Ki-67, MMP-2, MMP-9, and KDM5B decreased, and the apoptosis rate, Bax/Bcl-2, Cleaved Caspase 3/Caspase 3, Cleaved Caspase 7/Caspase 7, miR-212-3p and KDM5B mRNA increased (P<0.05). The effect of transfecting miR-212-3p mimics on MKN-45 cells was the same as butorphanol. Compared with the B-H+anti-miR-NC group, the B-H+anti-miR-212-3p group had higher number of Edu positive cells, invasion rate, and scratch healing rate, higher Ki-67, MMP-2, MMP-9, and KDM5B protein levels, and lower apoptosis rate, Bax/Bcl-2, Cleaved Caspase 3/Caspase 3, Cleaved Caspase 7/Caspase 7, miR-212-3p and KDM5B mRNA (P<0.05). Butorphanol can inhibit the proliferation, migration, and invasion of gastric cancer cells and promote apoptosis by upregulating miR-212-3p and suppressing KDM5B.
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