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Lycorine hydrochloride regulates the malignant biological behaviors of liver cancer HCCLM3 cells through the circASH2L/miR-124-3p axis
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Abstract:
Objective: To explore the effect of lycorine hydrochloride (LH) on the malignant biological behaviors of liver cancer HCCLM3 cells and its regulation on circASH2L/miR-124-3p axis. Methods: HCCLM3 cells treated with different concentrations of LH were divided into LH groups with different concentrations (LH-L group, LH-M group and LH-H group), Con group, si-NC group, si-circASH2L group, LH+pcDNA group, LH+pcDNA-circASH2L group. CCK-8 assay, plate clone formation test, flow cytometry, wound-healing assay, and Transwell assay were used to detect the proliferation, clone formation, apoptosis, migration and invasion of HCCLM3 cells. qPCR was used to detect the expression levels of circASH2L and miR-124-3p in HCCLM3 cells. Dual luciferase reporter gene assay was used to experiment detect the targeting relationship between circASH2L and miR-124-3p. WB was used to detect the expression of cleaved-caspase3, cleaved-caspase9, E-cadherin, and N-cadherin.Results: Compared with the Con group, the cell proliferation inhibition rates of LH groups with different concentrations were increased (P<0.05), the apoptosis rates and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the expression levels of miR-124-3p were increased (P<0.05), the number of clone formation and invasive cells was decreased (all P<0.05), the wound-healing rates and the expression levels of N-cadherin were decreased (all P<0.05), and the expression levels of circASH2L were decreased (P<0.05), and the difference among different concentration groups was statistically significant(all P<0.05). CircASH2L could negatively regulate miR-124-3p. Compared with the si-NC group, the cell proliferation inhibition rates of the si-circASH2L group was increased (P<0.05), and the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the wound-healing rate and the expression levels of N-cadherin were decreased (all P<0.05), and the number of clone formation and invasive cells was decreased (all P<0.05). Compared with the LH+pcDNA group, the expression level of miR-124-3p in the LH+ pcDNA-circASH2L group was decreased (P<0.05), the cell proliferation inhibition rate was decreased (P<0.05), the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were decreased (all P<0.05), the number of clone formation and invasive cells was increased (P<0.05), and the wound-healing rate and expression level of N-cadherin were increased (all P<0.05).
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Project Supported:
Project supported by the Science and Technology Breakthrough Plan of Henan Province（No.162102310331），and the Key Science and Technology Breakthrough Plan of Henan Province（No.142102310128）
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