摘要
目的探讨METTL14通过调控巨噬细胞分化抑制宫颈癌病理性发展及相关机制。方法检测宫颈癌病变样本METTL14 m RNA和蛋白,以及IL-6、iNOS、Arg-1和CD206表达变化。PMA诱导THP-1细胞转化为巨噬细胞,慢病毒过表达或抑制METTL14表达,检测IL-6、iNO、Arg-1和CD206表达变化以及PI3K/AKT/GSK3β/β-catenin信号通路相关蛋白表达情况。随后加入PI3K/AKT/GSK3β/β-catenin信号通路激动剂和抑制剂,检测过表达或抑制METTL14后,巨噬细胞IL-6、iNO、Arg-1和CD206表达变化,并取其上清制成条件培养基,孵育Hela细胞,检测细胞凋亡和增殖情况。结果1)宫颈癌病变组织中METTL14 mRNA和蛋白表达降低(P<0.05),巨噬细胞M1型标志物IL-6和iNOS表达明显降低(P<0.05),而M2型标志物Arg-1和CD206表达明显升高(P<0.05)。2)巨噬细胞过表达METTL14后,IL-6和iNOS表达明显升高(P<0.05),而Arg-1和CD206表达明显降低(P<0.05),M1/M2比例升高;抑制METTL14表达后,M1型标志物降低(P<0.05),M2型标志物升高(P<0.05),M1/M2比例降低。3)巨噬细胞中转染OE-METTL14慢病毒组PI3K/AKT/GSK3β/β-catenin信号通路被抑制(P<0.05);加入PI3K/AKT激动剂后,M1型标志物降低而M2型标记物升高(P<0.05),M1/M2比例降低;OE-METTL14可逆转此趋势。Sh-METTL14慢病毒组PI3K/AKT/GSK3β/β-catenin信号通路被激活(P<0.05),加入PI3K/AKT抑制剂后,M1型标志物升高而M2型标记物降低(P<0.05),M1/M2比例升高;Sh-METTL14可逆转此趋势。4)取转染OE-METTL14慢病毒后的巨噬细胞上清培养Hela细胞,可见细胞凋亡明显增多(P<0.05),增殖明显减少(P<0.05)。Sh-METTL14组的Hela细胞则表现出细胞凋亡减少(P<0.05),增殖增多(P<0.05)。结论METTL14通过PI3K/AKT/GSK3β/β-catenin信号通路调控巨噬细胞分化可能有促进宫颈癌细胞凋亡,抑制增殖的作用。
This study was designed to investigate the effect of METTL14 inhibiting cervical cancer pathological development by regulating macrophage differentiation and to explore the related mechanisms.The expression of METTL14 mRNA and protein,IL-6,iNOS,Arg-1 and CD206 in cervical cancer samples were detected.THP-1 cells were induced with PMA to transform into macrophages;lentivirus was used to overexpress or inhibit METTL14 expression,and the expression changes of IL-6,iNO,Arg-1 and CD206,as well as the expression of PI3K/AKT/GSK3β/β-catenin signal pathway related proteins were detected.Subsequently,PI3K/AKT/GSK3β/β-catenin signaling pathway agonists and inhibitors were added to detect the expression changes of IL-6,iNO,Arg-1and CD206 in macrophages after overexpression or inhibition of METTL14.The supernatant was used to prepare conditioned medium,which then applied to incubateHela cells.Then the apoptosis and proliferation of Helacells were detected.Data showed that the expression of METTL14 m RNA and protein in cervical cancer tissues decreased(P<0.05),and the expressions of IL-6 and i NOSas M1 markers of macrophages were decreased significantly(P<0.05),while the expressions of M2 markers Arg-1and CD206 increased significantly(P<0.05).After overexpression of METTL14 in macrophages,the expression ofIL-6 and i NOS increased significantly(P<0.05),while the expression of Arg-1 and CD206 decreased significantly(P<0.05),and the ratio of M1/M2 increased;After inhibition of METTL14 expression,M1 marker decreased(P<0.05),M2 marker increased(P<0.05),and M1/M2 ratio decreased.PI3K/AKT/GSK3β/β-catenin signal pathway wasinhibited in macrophages transfected with OE-METTL14 lentivirus(P<0.05).When PI3K/AKT agonist was added,M1 marker decreased while M2 marker increased(P<0.05),and the ratio of M1/M2 decreased.OE-METTL14reversed this trend.In Sh-METTL14 lentivirus group,PI3K/AKT/GSK3β/β-catenin signal pathway was activated(P<0.05).After PI3K/AKT inhibitor was added,M1 marker increased,while M2 marker decreased(P<0.05),and M1/M2 ratio increased.Sh-METTL14 reversed this trend.As for the Hela cells cultured in the supernatant ofmacrophages transfected with OE-METTL14 lentivirus,the apoptosis was significantly increased(P<0.05)andproliferation was significantly reduced(P<0.05).Hela cells in Sh-METTL14 group showed decreased apoptosis(P<0.05)and increased proliferation(P<0.05).In conclusion,METTL14 regulates macrophage differentiation throughPI3K/AKT/GSK3β/β-catenin signal pathway,which may promote apoptosis and inhibit proliferation of cervicalcancer cells.
作者
孟敬一
王娜
孙晓勤
MENG Jingyi;WANG Na;SUN Xiaoqin(Department of Gynecology,Zibo Maternal and Child Health Care Hospital,Zibo 255000,China;Department of Pathology,Zibo Maternal and Child Health Care Hospital,Zibo 255000,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第1期45-52,共8页
Immunological Journal
