YTH结构域家族蛋白2对砷诱导皮肤角质细胞恶性表型的影响  

Effects of YTH domain family protein 2 on malignant phenotype of arsenic-induced skin keratinocytes

作　　者：张乾 满瑾 孙东雷 赵田禾 张遵真 ZHANG Qian;MAN Jin;SUN Dong-lei;ZHAO Tian-he;ZHANG Zun-zhen(Department of Occupational and Environmental Health,West China School of Public Health and West China Fourth Hospital,Sichuan University,Chengdu,Sichuan,610041,China)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院劳动卫生与环境卫生学系,四川成都610041

出　　处：《现代预防医学》2023年第9期1694-1699,1728,共7页Modern Preventive Medicine

基　　金：国家自然科学基金(82073510)。

摘　　要：目的探究YTH结构域家族蛋白2(YTH domain family protein 2,YTHDF2)对砷诱导皮肤角质细胞恶性表型的作用,为砷致皮肤癌的机制提供新视角。方法以正常皮肤角质HaCat细胞和1μM亚砷酸钠连续染毒50代的恶性转化HaCat细胞(HaCat-T)为研究对象,采用免疫印迹实验检测YTHDF2蛋白在HaCat-T和HaCat细胞的表达水平。siRNA沉默HaCat-T细胞YTHDF2基因表达,CCKc-8实验、平板划痕愈合实验、Transwell实验以及流式细胞术检测YTHDF2敲低后HaCat-T细胞增殖、迁移、侵袭能力以及凋亡水平的变化,采用t检验分析组间差异。结果相比于HaCat细胞,HaCat-T细胞YTHDF2蛋白表达水平增高1.37倍(t=12.238,P<0.001)。siRNA干扰有效降低了HaCat-T细胞中44.40%(t=6.423,P=0.003)YTHDF2蛋白表达;敲低YTHDF2后,HaCat-T细胞增殖能力相比于未敲低的HaCat-T细胞在24、48和72 h分别降低32.17%(t=7.980,P=0.001)、35.18%(t=6.928,P=0.002)和41.61%(t=10.910,P<0.001);划痕实验表明YTHDF2敲低组的HaCat-T细胞相对迁移能力降低为未敲低组细胞的47.9%(t=6.288,P=0.003);在Transwell实验中,YTHDF2敲低组细胞的HaCat-T细胞穿膜数量为(114.7±9.96)个,显著低于对照组细胞[(213.3±16.51)个,t=5.119,P=0.007]。此外,YTHDF2敲低组HaCat-T细胞凋亡率为(39.66±0.41)%,明显高于未敲低组[(19.21±1.23)%,t=15.760,P<0.001]。结论YTHDF2可以促进砷诱导的皮肤角质细胞的增殖、迁移、侵袭能力和抑制细胞凋亡水平,可能是砷诱导皮肤角质细胞发生恶性转化的重要调控因子。Objective To investigate the role and mechanism of YTH domain family protein 2(YTHDF2)in malignant phenotype of arsenic-induced keratinocytes and provide a new perspective on the mechanism of arsenic-induced skin cancer.Methods Human-derived normal keratinous HaCat cells and malignant transformed HaCat cells(HaCat-T)stained with 1μM sodium arsenite for 50 consecutive generations were used for the study.Western blot assay was performed to detect the expression levels of YTHDF2 protein in HaCat-T and HaCat cells.Small interfering RNA(siRNA)technology silenced YTHDF2 gene expression in HaCat-T cells,and CCK-8 assay,wound healing assay,Transwell assay,and flow cytometry were used to detect changes in proliferation,migration,invasion,and apoptosis levels of malignant transformed cells after YTHDF2 knockdown.The differences between groups were analyzed using the t-test.Results YTHDF2 protein expression in HaCat-T cells was increased 1.37-fold(t=12.238,P<0.001)compared with HaCat cells.The siRNA intervention effectively reduced YTHDF2 protein expression in HaCat-T cells by 44.40%,with significant difference(t=6.423,P=0.003).The proliferation of HaCat-T cells after knockdown of YTHDF2 was reduced by 32.17%(t=7.980,P=0.001),35.18%(t=6.928,P=0.002),and 41.61%(t=10.910,P<0.001)at 24 h,48 h,and 72 h,respectively,compared with sicontrol HaCat-T cells,with statistically significant differences.The wound healing assay showed that the relative migration ability of HaCat-T cells in the YTHDF2 knockdown cells were reduced by 47.9%(t=6.288,P=0.003).The transwell assay showed that the number of membrane penetration in the YTHDF2 knockdown group was(114.7±9.96),which was lower than that in the non-knockdown group(213.3±16.51,t=5.119,P=0.007).Flow cytometry results showed that the apoptosis rate of YTHDF2 knockdown cells was(39.66±0.41)%,which was significantly higher than that of the non-knockdown group[(19.21±1.23)%,t=15.760,P<0.001].Conclusion YTHDF2 can promote proliferation,migration,and invasion and inhibit apoptosis levels

关 键 词：YTHDF2 砷 人源性皮肤角质细胞 细胞恶性转化 

分 类 号：R114[医药卫生—卫生毒理学]