Effects of circular RNA CDR1-AS on proliferation,apoptosis and immune factor expression of lung cancer cells by targeting miR-1277
Objective:To investigate the effect and possible mechanism of circular RNA(circRNA)cerebellar degeneration-related protein 1 antisense transcript(CDR1-AS)on proliferation,apoptosis and immune factors expressions of lung cancer cells.Methods:Using BEAS-2B in adjacent tissues and bronchial epithelial cells as controls,qRT-PCR was used to detect expressions of CDR1-AS and miR-1277 in lung cancer tissues and lung cancer cell lines(NCI-H23,H1299,NCI-H446).Taking lung cancer H1299 cells as research object,after they were transfected with CDR1-AS small interfering RNA(si-CDR1-AS)or miR-1277 mimic,or co-transfected with si-CDR1-AS and miR-1277 inhibitor,CCK-8 experiment,clone formation experiment and flow cytometry were used to detect cell viability,clone formation number and apoptosis rate,respectively.ELISA was used to detect expressions of immune factors(TNF-α,IFN-γ)in cell culture supernatant.Dual-luciferase reporter gene experiment was used to verify the regulatory relation-ship between CDR1-AS and miR-1277.Results:Compared with adjacent tissues or BEAS-2B cells,expression of CDR1-AS in lung cancer tissues or cell lines(NCI-H23,H1299,NCI-H446)was increased(P<0.05),while expression of miR-1277 was decreased(P<0.05).After silencing CDR1-AS or overexpressing miR-1277,activity of H1299 cells,the number of clones and the secretion of TNF-α were decreased(P<0.05),while apoptosis rate and secretion of IFN-γ were increased(P<0.05).CDR1-AS targeted and bound to miR-1277,and expression of miR-1277 was increased in H1299 cells that silenced CDR1-AS(P<0.05).Down-regulation of miR-1277 reversed effects of silencing CDR1-AS on proliferation,apoptosis and expressions of immune factors in H1299 cells.Conclu-sion:Expression of CDR1-AS is up-regulated in lung cancer tissues and cell lines,silencing its expression can inhibit proliferation and tumor cell immune escape of lung cancer H1299 cells,and induce cell apoptosis by targeting up-regulation of miR-1277.
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