LncTUG1通过靶向miR-212-3p对口腔鳞状细胞癌细胞NK细胞杀伤敏感性的影响

Effect of LncTUG1 on NK cell killing sensitivity in oral squamous cell carcinoma cells by targeting miR-212-3p

目的:探讨长链非编码RNA(Lnc)牛磺酸调节基因1(taurine up-regulated gene 1,TUG1)对口腔鳞状细胞癌细胞自然杀伤细胞(nature killer cell,NK)杀伤敏感性的影响。方法:以口腔鳞状细胞癌细胞作为体外实验对象,转染TUG1 siRNA,利用qRT-PCR、MTT、流式细胞术、LDH方法,分别检测TUG1表达量、细胞增殖、细胞凋亡和NK细胞杀伤率。利用生物信息学软件预测TUG1与miR-212-3p靶向互补结合,构建荧光素酶报告载体,鉴定靶向关系。在口腔鳞状细胞癌细胞中共转染TUG1 siRNA和miR-212-3p抑制剂,评估miR-212-3p抑制剂对TUG1 siRNA影响口腔鳞状细胞癌细胞增殖凋亡和NK细胞杀伤敏感性的影响。采用SPSS 21.0软件包对实验数据进行统计学分析。结果:TUG1 siRNA可显著降低口腔鳞状细胞癌细胞中TUG1的表达水平(P=0.000),降低细胞增殖能力(P=0.001),促进细胞凋亡(P=0.000),增加NK细胞杀伤率(P<0.01)。TUG1 siRNA靶向提高miR-212-3p表达量。miR-212-3p抑制剂可逆转TUG1 siRNA对口腔鳞状细胞癌细胞增殖、凋亡和NK细胞杀伤率的影响。结论:下调TUG1可靶向负调控miR-212-3p,抑制口腔鳞状细胞癌细胞增殖并诱导凋亡,提高NK细胞杀伤敏感性。

PURPOSE:To investigate the effect of LncTUG1 on NK cell killing sensitivity in oral squamous cell carcinoma cells.METHODS:Oral squamous cell carcinoma cells were used as experimental objects in vitro.TUG1 siRNA was transfected,the expression of TUG1,cell proliferation,cell apoptosis and NK cell killing rate were detected by qRT-PCR,MTT,flow cytometry and LDH.Bioinformatics software was used to predict that TUG1 and miR-212-3p will target and complement each other,so luciferase reporter vector was constructed and the targeting relationship was identified.TUG1 siRNA and miR-212-3p inhibitor were co-transfected into oral squamous cell carcinoma cells,the effects of miR-212-3p inhibitor on TUG1 siRNA on proliferation,apoptosis and NK cell killing sensitivity of oral squamous cell carcinoma cells were evaluated.The data were analyzed with SPSS 21.0 software package.RESULTS:TUG1 siRNA could significantly reduce the expression of TUG1 in oral squamous cell carcinoma cells(P=0.000),decrease cell proliferation(P=0.001),promote cell apoptosis(P=0.000),increase the killing rate of NK cells(P<0.01).TUG1 siRNA targeted to increase the expression of miR-212-3p.miR-212-3p inhibitor could reverse the effects of TUG1 siRNA on proliferation,apoptosis and NK cell killing rate of oral squamous cell carcinoma cells.CONCLUSIONS:Down-regulation of TUG1 targeting and negative regulation of miR-212-3p inhibits proliferation,induces apoptosis and improves NK cell killing sensitivity of oral squamous cell carcinoma cells.