CCAT1 targeted inhibition of miR-375-3p on apoptosis of human oral cancer SAS cells via mitochondrial pathway
Objective:To explore the role of long chain non-coding RNA(lncRNA)colon cancer-related transcription factor 1(CCAT1)targeting miR-375-3p in inducing the apoptosis of human oral cancer SAS cells through mitochondrial pathway.Methods:Human oral cancer cell line SAS cells were randomly divided into blank group,CCAT1 down-regulated group,CCAT1 up-regulated group,CCAT1 down-regulated control group,CCAT1 up-regulated control group,miR-375-3p down-regulated group,miR-375-3p up-regulated group,miR-375-3p down-regulated control group,and miR-375-3p up-regulated control group.After 48 hours of transfection,the apoptosis was detected by flow cytometry.RT-qPCR was used to detect the expression of miR-375-3p and the mRNA expressions of genes related to mitochondrial apoptosis pathway,such as caspase-3,caspase-9,Bcl-2,and Bcl-2-related X protein(Bax).The protein expressions of caspase-3,caspase-9,Bcl-2 and Bax were detected by Western blotting.The targeting relationship between CCAT1 and miR-375-3p was analyzed by dual luciferase reporter assay.Results:Compared with the blank group and the CCAT1 down-regulated control group,the apoptosis rate,miR-375-3p expression,and mRNA and protein expressions of caspase-3,caspase-9,and Bax in the CCAT1 down-regulated group were increased,while the Bcl-2 mRNA and protein expressions were decreased.Compared with the blank group and CCAT1 down-regulated control group,the apoptosis rate,miR-375-3p expression,and mRNA and protein expressions of caspase-3,caspase-9,and Bax in the CCAT1 up-regulated group were decreased,while the Bcl-2 mRNA and protein expressions were increased.Compared with the blank group and the miR-375-3p down-regulated control group,the apoptosis rate,the expression of miR-375-3p,and mRNA and protein expressions of caspase-3,caspase-9,and Bax in the miR-375-3p down-regulated group were decreased,while Bcl-2 mRNA and protein expressions were increased.Compared with the blank group and the miR-375-3p up-regulated control group,the apoptosis rate,the expression of miR-375-3p,the mRNA and protein expressions of caspase-3,caspase-9,Bax were increased in the miR-375-3p up-regulated group,while the Bcl-2 mRNA and protein expressions were decreased.The dual luciferase reporter assay confirmed that CCAT1 could target miR-375-3p.Conclusion:CCAT1 can inhibit the apoptosis of human oral cancer SAS cells,which is speculated to be achieved by targeting the expression of miR-375-3p and regulating the expressions of genes and proteins related to the mitochondrial apoptosis pathway.
long chain non-coding RNAcolon cancer related transcription factor 1miR-375-3poral cancermitochondrial pathwayapoptosis
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