LncRNA MEG8 mediates immune escape in colorectal cancer cells by regulating MICA/B via targeting miR-15a-5p
Objective:To explore mechanism of long non-coding RNA maternally expressed gene 8(LncRNA MEG8)mediates immune escape in colorectal cancer(CRC)cells by regulating MICA/B via targeting miR-15a-5p.Methods:RT-qPCR and Western blot were used to detect expressions of MEG8,miR-15a-5p,MICA,MICB,NKG2D protein in CRC tissue and cell lines;dual luciferase experiment was used to verify regulatory relationship of MEG8 on miR-15a-5p;NC,MEG8,miR-NC and miR-15a-5p were respectively or co-transfected into SW480 and SW620 cells by liposome transfection,and recorded as NC group,MEG8 group,MEG8+miR-NC group,MEG8+miR-15a-5p group;NK cells were cocultured with SW480 and SW620 cells respectively;ELISA was used to detect TNF-α and IFN-γ levels in coculture medium;CCK-8,EdU staining,Transwell assay were used to detect cell prolifera-tion,migration and invasion ability;RT-qPCR and Western blot were used to detect MEG8,miR-15a-5p,MICA,MICB,NKG2D protein levels in cells.Results:Compared with adjacent cancer tissues or normal colon epithelial cells,MEG8,MICA,MICB,NKG2D mRNA and protein levels in CRC tissues and cell lines were decreased,while miR-15a-5p level was increased(P<0.05).MEG8 targeting regulated miR-15a-5p.In cocultivation system,compared with NC group,MICA,MICB,NKG2D protein levels,TNF-α,IFN-γ levels in cocultured supernatant in MEG8 group were significantly increased,after 1 d,2 d,and 3 d of cell culture,OD value,EdU positive rate,number of migrating and invading cells were significantly decreased(P<0.05);overexpression of miR-15a-5p could partially reverse effects of overexpression of MEG8 on MICA,MICB,NKG2D,TNF-α,IFN-γ levels in cells,prolifera-tion,invasion and metastasis ability(P<0.05).Conclusion:MEG8 promotes NK cell activity and inhibits CRC cell immune escape by regulating MICA and MICB expressions via targeting miR-15a-5p.
Long non-coding RNA maternally expressed gene 8miR-15a-5pColorectal cancerImmune escapeMHC classⅠ chain-related protein A/B
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