METTL3介导的m^(6)A修饰调控miR-1224-5p在前列腺癌细胞增殖和迁移中的作用  

Role of METTL3-mediated m^(6)A modification in regulating miR-1224-5p in the proliferation and migration of prostate cancer cells

作　　者：胡东来 赵梓岐 楚元奎[1,2] HU Donglai;ZHAO Ziqi;CHU Yuankui(School of Laboratory Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia,China;Medical Experimental Center,General Hospital of Ningxia Medical University,Yinchuan 750003,Ningxia,China)

机构地区：[1]宁夏医科大学检验学院,宁夏银川750004 [2]宁夏医科大学总医院医学实验中心,宁夏银川750003

出　　处：《中国肿瘤生物治疗杂志》2025年第1期64-72,共9页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金(No.82160465)。

摘　　要：目的:探究miR-1224-5p在前列腺癌细胞增殖、迁移与凋亡中的生物学作用及其表达调控机制。方法:选用人前列腺癌细胞PC3、DU145、LNCaP、22Rv1和正常前列腺上皮细胞RWPE-1,利用qPCR法检测miR-1224-5p在前列腺癌细胞中的表达。使用脂质体转染技术分别将miR-1224-5p模拟物(mimic)、抑制剂(inhibitor)及相应的阴性对照(NC)质粒转染至PC3和DU145细胞中,qPCR法验证转染效率,采用CCK-8实验、平板克隆形成实验、划痕愈合实验和流式细胞术分别检测转染miR-1224-5p mimic与inhibitor对细胞增殖、迁移和凋亡的影响。借助SRAMP网站预测pri-miR-1224-5p序列中潜在的N6-甲基腺苷(m^(6)A)修饰位点,通过甲基化RNA免疫沉淀实验(MeRIP)验证预测结果,qPCR法检测m^(6)A修饰关键调控分子甲基转移酶3(METTL3)在前列腺癌细胞中的表达,CCK-8实验和Transwell实验分别检测转染METTL3 siRNA对PC3和DU145细胞增殖和迁移的影响,qPCR法和MeRIP检测METTL3介导的m^(6)A修饰对miR-1224-5p表达的调控作用。结果:miR-1224-5p在前列腺癌细胞中均呈高表达(均P<0.01)。转染miR-1224-5p mimic能够促进PC3和DU145细胞增殖、迁移并抑制细胞凋亡(P<0.05或P<0.01),转染miR-1224-5pinhibitor能够抑制PC3和DU145细胞增殖、迁移并诱导细胞凋亡(P<0.05或P<0.01)。pri-miR-1224-5p具有m^(6)A修饰位点;METTL3在前列腺癌细胞中均呈高表达(均P<0.01),转染METTL3 siRNA能够抑制PC3和DU145细胞的增殖和迁移(均P<0.01);METTL3介导的m^(6)A修饰能够调控miR-1224-5p的表达。结论:miR-1224-5p受METTL3介导的m^(6)A修饰调控,从而在前列腺癌细胞中呈高表达,下调miR-1224-5p能够抑制前列腺癌细胞的增殖、迁移并诱导细胞凋亡。Objective:To investigate the biological role of miR-1224-5p in the proliferation,migration and apoptosis of prostate cancer cells,as well as its regulatory mechanism of expression.Methods:Human prostate cancer cells(PC3,DU145,LNCaP,22Rv1)and normal prostate epithelial RWPE-1 cells were selected for this study.The expression of miR-1224-5p in prostate cancer cells was detected by qPCR.miR-1224-5p mimic,inhibitor and corresponding negative control(NC)plasmids were transfected into PC3 and DU145 cells by liposome transfection technology,respectively.The transfection efficiency was verified by qPCR.The effects of miR-1224-5p mimic and inhibitor on cell proliferation,migration and apoptosis were detected by CCK-8 assay,plate clone formation assay,scratch healing assay and flow cytometry.The potential N6-methyladenosine(m^(6)A)modification sites in the pri-miR-1224-5p sequence were predicted using SRAMP website,and the prediction results were verified using methylated RNA immunoprecipitation(MeRIP).The expression of methyltransferase 3(METTL3),a key methyltransferase involved in m^(6)A modification,in prostate cancer cells was detected by qPCR.CCK-8 assay and Transwell assay were used to detect the effect of METTL3 siRNA transfection on the proliferation and migration of PC3 and DU145 cells.The regulatory effect of METTL3-mediated m^(6)A modification on the expression of miR-1224-5p was detected by qPCR and MeRIP.Results:miR-1224-5p was upregulated in prostate cancer cells(all P<0.01).Transfection with miR-1224-5p mimic promoted the proliferation,migration and inhibited the apoptosis of PC3 and DU145 cells(P<0.05 or P<0.01).Conversely,miR-1224-5p inhibitor suppressed the proliferation,migration and induced apoptosis of PC3 and DU145 cells(P<0.05 or P<0.01).pri-miR-1224-5p contained m^(6)A modification sites.METTL3 was highly expressed in prostate cancer cells(all P<0.01).Transfection of METTL3 siRNA inhibited the proliferation and migration of PC3 and DU145 cells(all P<0.01).METTL3-mediated m^(6)A modification regulate

关 键 词：前列腺癌 miR-1224-5p N^(6)-甲基腺苷 甲基转移酶3 增殖 迁移 凋亡 

分 类 号：R737.25[医药卫生—肿瘤] R730.2[医药卫生—临床医学]