Effects of Fusobacterium nucleatum and miR-497 on proliferation,invasion and migration of colorectal cancer cells and their regulatory relationships
Objective To observe the effects of Fusobacterium nucleatum and microRNA-497(miR-497)on the prolif-eration,invasion and migration of colorectal cancer cells,and to analyze their regulatory relationships.Methods Colorec-tal cancer cell line HCT116 was transfected with miR-497 overexpression plasmid miR-497 mimic and was randomly divid-ed into the infection index(MOI)100 group,MOI 200 group,MOI 500 group,and MOI 1 000 group.They were co-cul-tured with Fusobacterium nucleatum with MOI of 100,200,500 and 1 000,respectively.Real-time fluorescent quantita-tive PCR was used to detect miR-497 in each group.The treatment concentration of the bacteria with the lowest expression of miR-497 was selected as the subsequent action concentration of Fusobacterium nucleatum.HCT116 cells were randomly divided into the control group,miR-497 over-expression group,miR-497 down-expression group,miR-497 over-expres-sion negative control group and miR-497 down-expression negative control group,respectively.Cells in the control group were cultured without treatment.Cells in the miR-497 over-expression group,miR-497 down-expression group,over-ex-pression negative control group,and down-expression negative control group were transfected with miR-497 mimics,miR-497 inhibitor,mimic NC,and inhibitor NC,respectively,by Lipofectamine 2000.Cell proliferation ability was observed by CCK-8 assay,cell invasion ability was observed by Transwell assay,and cell migration ability was observed by Scratch healing assay.HCT116 cells were randomly divided into the control group,miR-497 over-expression group,overe-xpres-sion negative control group,and over-expression+Fusobacterium nucleatum group,respectively.Cells in the control group were cultured without treatment,and cells in the miR-497 over-expression group were transfected with miR-497 mim-ic,while cells in the over-expression negative control group were transfected with mimic NC.After transfecting miR-497 mimic,cells in the overexpression+Fusobacterium nucleatum group were co-cultured with selected concentration of Fuso-bacterium nucleatum.The cell proliferation ability was observed by CCK-8 assay,the cell invasion ability was observed by Transwell assay,and the cell migration ability was observed by Scratch healing assay.Results The miR-497 expression was as follows:MOI 100 group>MOI 200 group>MOI 500 group>MOI 1 000 group(all P<0.05),and MOI 1 000 was se-lected as the subsequent action concentration of Fusobacterium nucleatum.Cell proliferation rate,the number of invasive cells and migration rate were as follows:miR-497 down-expression group>control group,overexpression negative control group,down-expression negative control group>miR-497 over-expression group(all P<0.05).Cell proliferation rate,the number of invasive cells,and migration rate were as follows:miR-497 overexpression group<overexpression+Fusobacte-rium nucleatum group<control group,overexpression negative control group(all P<0.05).Conclusion Fusobacterium nucleatum can promote the proliferation,invasion and migration of colorectal cancer cells through negative regulation of miR-497.
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