miR-145-5p抑制造釉细胞型颅咽管瘤细胞增殖、侵袭和SOX9/β-catenin表达  

miR-145-5p inhibits proliferation,invasion and SOX9/β-catenin expression of adamantinomatous craniopharyngioma cells

作　　者：夏可顺 黄耀辉 王茂 刘志勇[2] 谭博 宋鹏 印晓鸿 Keshun Xia;Yaohui Huang;Mao Wang;Zhiyong Liu;Bo Tan;Peng Song;Xiaohong Yin(Department of Neurosurgery,Guangyuan Central Hospital Affiliated to North Sichuan Medical College,Guangyuan 628099,China;Department of Neurosurgery,West China Hospital,Sichuan University,Chengdu 610044,China)

机构地区：[1]川北医学院附属广元市中心医院神经外科,四川广元628099 [2]四川大学华西医院神经外科,成都610044

出　　处：《中华神经创伤外科电子杂志》2024年第1期6-15,共10页Chinese Journal Of Neurotraumatic Surgery：Electronic Edition

基　　金：四川省临床重点专科建设项目(2022)。

摘　　要：目的探究miR-145-5p对造釉细胞型颅咽管瘤(aCP)细胞增殖、侵袭和凋亡的影响及作用机制。方法选取2019年9月至2022年9月在四川大学华西医院和广元市中心医院神经外科术中切除的aCP和乳头状颅咽管瘤(pCP)各5例。采用实时荧光定量PCR(qRT-PCR)检测aCP和pCP中miR-145-5p、SOX9、CTNNB1(β-catenin基因)mRNA的表达差异,采用免疫荧光及Western blot检测SOX9、β-catenin蛋白的表达;通过CCK8实验、Transwell小室及流式细胞术分别检测颅咽管瘤细胞的增殖、侵袭和凋亡水平。将aCP细胞分为aCP组(空白对照,未转染)、aCP+miR-145-5p control组(阴性对照)、aCP+miR-145-5p mimic组(过表达),研究miR-145-5p对aCP细胞的增殖、侵袭和凋亡及SOX9、β-catenin表达的影响。结果与pCP相比,aCP细胞中miR-145-5p mRNA表达明显降低,SOX9、CTNNB1 mRNA、SOX9及胞核β-catenin蛋白表达明显增高,胞质β-catenin蛋白表达略降低,细胞增殖和侵袭显著增强,差异均有统计学意义(P<0.05)。aCP组、aCP+miR-145-5p control组和aCP+miR-145-5p mimic组在miR-145-5p、SOX9、CTNNB1 mRNA,SOX9、胞核β-catenin、胞质β-catenin蛋白表达以及细胞增殖和侵袭方面比较,差异均有统计学意义(P<0.05)。两两比较显示,aCP+miR-145-5p mimic组较其他2组miR-145-5p mRNA表达显著升高,细胞增殖和侵袭减弱,凋亡增加,SOX9、CTNNB1 mRNA表达明显降低;aCP+miR-145-5p mimic组较aCP+miR-145-5p control组SOX9、胞核β-catenin蛋白表达明显降低,胞质β-catenin蛋白表达略升高,差异均有统计学意义(P<0.05)。结论miR-145-5p可能通过下调SOX9、β-catenin抑制aCP的增殖、侵袭能力,促进细胞凋亡。Objective To investigate the effects of miR-145-5p on the proliferation,invasion,and apoptosis of adamantinomatous craniopharyngioma(aCP)cells and its mechanism of action.Methods Five cases of aCP and five cases of papillary craniopharyngioma(pCP)resected at Neurosurgery Department of West China Hospital of Sichuan University and Guangyuan Central Hospital from September 2019 to September 2022 were selected.The expression differences of miR-145-5p,SOX9,and CTNNB1(β-catenin gene)mRNA in aCP and pCP were detected by real-time fluorescence quantitative PCR(qRT-PCR).The expression of SOX9 and β-catenin proteins were detected by immunofluorescence and Western blot.The proliferation,invasion,and apoptosis levels of craniopharyngioma cells were measured through CCK8 assay,Transwell chamber,and flow cytometry,respectively.aCP cells were divided into aCP group(blank control,untransfected),aCP+miR-145-5p control group(negative control),and aCP+miR-145-5p mimic group(overexpression)to study the effects of miR-145-5p on the proliferation,invasion,and apoptosis of aCP cells and on the expression of SOX9 and β-catenin.Results Compared with pCP,aCP cells showed significantly reduced expression of miR-145-5p mRNA and increased expression of SOX9、CTNNB1 mRNA,SOX9、nuclearβ-catenin proteins,and a decreased in cytoplasmic β-catenin protein expression,concurrently,cell proliferation and invasion were significantly enhanced,with all differences being statistically significant(P<0.05).There were differences among the aCP group,aCP+miR-145-5p control group,and aCP+miR-145-5p mimic group in terms of miR-145-5p,SOX9,and CTNNB1 mRNA expression,as well as SOX9,nuclear β-catenin,and cytoplasmic β-catenin protein expression,and in cell proliferation and invasion(P<0.05).Pairwise comparisons between groups showed that the aCP+miR-145-5p mimic group had significantly increased expression of miR-145-5p mRNA,weakened proliferation and invasion,and increased apoptosis compared to the other two groups.The expression of SOX9 and CTNNB1 mR

关 键 词：造釉细胞型颅咽管瘤 乳头状颅咽管瘤 miR-145-5p SOX9 Β-CATENIN 增殖 侵袭 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]