摘要
目的:探讨外泌体分泌miR-133a-3p调控TGFBR1从而促进前列腺癌进展的分子机制。方法:提取前列腺癌细胞中的外泌体。免疫印迹实验(Western blot,WB)实验检测外泌体相关标记蛋,如白细胞分化抗原81(CD81)和白细胞分化抗原63(CD63)的表达;miRNA矩阵测序检测miR-133a-3p的表达;细胞克隆实验、细胞增殖8(CCK-8)实验、EDU细胞增殖实验、细胞迁移实验检测过表达miR-133a-3p对前列腺癌增殖和转移能力的影响;双荧光素酶报告基因实验检测miR-133a-3p结合转化生长因子β受体1(transforming growth factorβreceptor 1,TGFBR1)的情况;实时荧光定量逆转录聚合酶链反应(qRT-PCR)和WB检测调控miR-133a-3p后对TGFBR1表达的影响。结果:相较于生理盐水(NC)组,共培养外泌体的CD81和CD63表达升高(P<0.05)。细胞克隆实验、CCK-8实验、EDU实验、划痕、细胞迁移实验提示,相较于NC组,转染miR-133a mimic模拟物组(miR-133a-3p组)的细胞增殖、转移能力下降(P<0.05)。双荧光素酶报告基因实验显示,miR-133a-3p结合TGFBR1序列,且过表达miR-133a-3p后,抑制了TGFBR1的表达(P<0.05)。WB结果显示,相较于NC组,si-TGFBR1组中的WNT、GSK3β、C-MYC和β-catenin的表达下降,oe-TGFBR1组中WNT、GSK3β、C-MYC和β-catenin的表达上升(P<0.05),而oe-TGFBR1+miR-133a-3p组中WNT、GSK3β、C-MYC和β-catenin的表达与NC组无差异。结论:前列腺癌细胞来源外泌体miR-133a-3p靶向调控TGFBR1从而促进前列腺癌细胞恶性表型。
Objective:To investigate the molecular mechanism by which exosomal secretion of miR-133a-3p regulates TGFBR1 and promotes the progression of prostate cancer.Methods:Exosomes were extracted from prostate cancer cells,and the expression of exosom-associated markers such as leukocyte differentiation antigen 81(CD81)and leukocyte differentiation antigen 63(CD63)was detected by Western blot assay(WB).The expression of miR-133a-3p was detected by miRNA matrix sequencing.Cell cloning experiment,cell proliferation 8(CCK-8)experiment,EdU cell proliferation experiment and cell migration experiment were used to detect the effects of co-cultured exosomes+miR-133a-3p on the proliferation and metastasis of prostate cancer.Dual luciferase reporter gene assay was used to detect the binding of miR-133a-3p to TGFBR1.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR)and WB were used to detect the effect of regulation of miR-133a-3p on transforming growth factorβreceptor 1(TGFBR1).Results:Compared with NC group,the expressions of CD81 and CD63 in co-cultured exosomes were increased(P<0.05).Cell cloning experiment,CCK-8 experiment,EDU experiment,scratch experiment,and cell migration experiment showed that compared with NC group,the cell proliferation and metastasis ability of Mir-133a-3P group(transfected with miR-133a mimic)decreased(P<0.05).Dual luciferase reporter gene experiment showed that miR-133a-3p binding TGFBR1 sequence and overexpression of miR-133a-3p inhibited the expression of TGFBR1(P<0.05).WB results showed that compared with NC group,the expressions of WNT,GSK3β,C-MYC andβ-catenin in si-TGFBR1 group were decreased,while the expressions of WNT,GSK3β,C-MYC andβ-catenin in oe-TGFBR1 group were increased(P<0.05).However,the expressions of WNT,GSK3β,C-MYC andβ-catenin in oe-TGFBR1+miR-133a-3p group were not different from those in NC group.Conclusion:Prostate cancer cell-derived exosome miR-133a-3p targets TGFBR1 to promote the malignant phenotype of prostate cancer cells.
作者
伍宏亮
汪盛
陈志军
杨帅
孙文衍
关翰
WU Hongliang;WANG Sheng;CHEN Zhijun;YANG Shuai;SUN Wenyan;GUAN Han(Department of Urology,The First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China)
出处
《包头医学院学报》
2025年第5期42-47,共6页
Journal of Baotou Medical College
基金
2023年度安徽省高校自然科学重点项目(2023AH051942)
2024年度安徽省高校自然科学重点项目(2024AH051278)
2019年度蚌埠医学院自然科学基金重点项目(BYKY2019088ZD)
2021年度蚌埠市级科技创新指导类项目(20210341)。
