miR-155诱导FSTL1基因对鼻咽癌细胞增殖、凋亡和免疫逃避的作用

Effects of miR-155 targeting FSTL1 gene on proliferation,apoptosis and immune evasion of nasopharyngeal carcinoma cells

目的探讨miR-155诱导FSTL1基因对鼻咽癌细胞增殖、凋亡和免疫逃避的作用。方法通过qRT-PCR检测人正常鼻咽上皮细胞NP69、鼻咽癌上皮细胞CNE1、CNE2、HONE1、HK-1和CNE2Z中miR-155和FSTL1的表达,取对数生长期的CNE2Z细胞进行实验,转染时共计分3组,分为miR-155 minics组、miR-155 inhibitor组和NC组,CCK-8检测细胞增殖,流式细胞术实验检测细胞凋亡,双荧光素酶报告实验检测靶向关系,Western blot检测FSTL1、C-caspase3、Bax、Bcl-2、ITCH和MAVS蛋白水平,酶联免疫吸附试验检测IFN-γ水平。结果与miR-155 inhibitor组相比,miR-155 minics组细胞增殖能力显著上升(P<0.05);miR-155 inhibitor组细胞凋亡率显著高于与NC组(P<0.05),miR-155 minics组细胞凋亡率明显低于miR-155 inhibitor组(P<0.05)。miR-155 inhibitor组FSTL1蛋白水平明显高于NC组(P<0.05),miR-155 minics组FSTL1蛋白水平显著低于miR-155 inhibitor组(P<0.05);miR-155 inhibitor组C-caspase3、Bax、Bcl-2蛋白水平明显低于NC组(P<0.05),miR-155 minics组C-caspase3、Bax、Bcl-2蛋白水平显著高于miR-155 inhibitor组(P<0.05)。FSTL1组中ITCH蛋白表达显著低于对照组(P<0.05);FSTL1组中MAVS蛋白表达显著高于对照组(P<0.05);FSTL1组中IFNγ水平明显高于对照组(P<0.05)。结论抑制miR-155表达能够降低鼻咽癌细胞的增殖能力,促进鼻咽癌细胞凋亡,miR-155可能通过诱导FSTL1基因参与鼻咽癌细胞的调控,同时通过诱导FSTL1基因,降低鼻咽癌细胞中ITCH表达,增加MAVS和IFNγ表达,达到机体内动态的免疫平衡。

Objective To investigate the effect of miR-155 targeting FSTL1 gene on proliferation,apoptosis and immune evasion of nasopharyngeal carcinoma cells.Methods The expression of miR-155 and FSTL1 in NP69,CNE1,CNE2,HONE1,HK-1 and CNE2Z cells were detected by qRT-PCR.CNE2Z cells in logarithmic growth phase were selected for experiment.The transfection was divided into three groups:miR-155 Minics,miR-155 inhibitor and NC groups.CCK-8 was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis,dual luciferase reporter assay was used to detect the targeting relationship.Western blot was used to detect the protein levels of FSTL1,c-caspase3,Bax,Bcl-2,itch and Mavs,and enzyme-linked immunosorbent assay was used to detect IFNγ.Results Compared with miR-155 inhibitor group,the proliferation ability of miR-155 Minics group was significantly increased(P<0.05);the apoptosis rate of miR-155 inhibitor group was significantly higher than that of NC group(P<0.05),and the apoptosis rate of miR-155 Minics group was significantly lower than that of miR-155 inhibitor group(P<0.05).The FSTL1 protein level in miR-155 inhibitor group was significantly higher than that in NC group(P<0.05),and the FSTL1 protein level in miR-155 Minics group was significantly lower than that in miR-155 Minics group.The protein levels of c-caspase3,Bax and Bcl-2 in miR-155 inhibitor group were significantly lower than those in NC group(P<0.05),and the protein levels of c-caspase3,Bax and Bcl-2 in miR-155 Minics group were significantly higher than those in miR-155 inhibitor group(P<0.05).The expression of ITCH protein in FSTL1 group was significantly lower than that in control group(P<0.05);the expression of Mavs protein in FSTL1 group was significantly higher than that in control group(P<0.05);the level of IFNγin FSTL1 group was significantly higher than that in control group(P<0.05).Conclusion Inhibiting the expression of miR-155 can reduce the proliferation of nasopharyngeal carcinoma cells and promote the apoptosis of nasopharyngeal carcinoma cells.MiR-155 may participate in the regulation of nasopharyngeal carcinoma cells by targeting FSTL1 gene.At the same time,by targeting FSTL1 gene,it can reduce the expression of itch in nasopharyngeal carcinoma cells and increase the expression of Mavs and IFNγ,so as to achieve the dynamic immune balance in the body.