脊髓小脑共济失调症相关蛋白Ataxin-3抑制张力蛋白同源基因通过蛋白激酶B/雷帕霉素靶蛋白信号通路促进睾丸癌细胞增殖的机制  

Ataxin-3 promotes proliferation of testicular cancer cells by inhibiting anti-oncogene PTEN and activates the AKT/mTOR signaling pathway:a mechanistic study

作　　者：史建华[1] 刘瑞强[1] 尹茂轩[1] 杨帮东[1] Shi Jianhua;Liu Ruiqiang;Yin Maoxuan;Yang Bangdong(Department of Urology,Fuyang People’s Hospital,Puyang,Henan 457000,China)

机构地区：[1]河南省濮阳市人民医院泌尿外科,457000

出　　处：《中华生物医学工程杂志》2019年第5期530-535,共6页Chinese Journal of Biomedical Engineering

摘　　要：目的探究脊髓小脑共济失调症相关蛋白(Ataxin)-3在睾丸癌(TC)组织中的表达及促进TC细胞增殖的机制。方法选取2015年1月至2019年5月在濮阳市人民医院泌尿外科接受睾丸切除术患者的TC组织及癌旁组织标本共38对,以免疫组化染色、实时荧光定量PCR检测Ataxin-3表达,以CCK8实验检测TC细胞增殖情况,并以免疫印迹检测Ataxin-3过表达或敲除后张力蛋白同源基因(PTEN)及蛋白激酶B/雷帕霉素靶蛋白(AKT/mTOR)信号通路相关蛋白水平。结果Ataxin-3蛋白和mRNA在TC组织中的表达水平明显高于癌旁组织(P<0.05),Ataxin-3表达与TC患者的年龄、病理类型无关(P>0.05),而与临床分期及转移明显相关(P<0.05)。当Ataxin-3蛋白在TCam-2细胞中过表达时可促进细胞增殖,而在TCam-2细胞中转染siRNA,敲除Ataxin-3后可抑制细胞增殖和侵袭。Ataxin-3过表达时TCam-2细胞中PTEN蛋白表达明显抑制,而敲除后PTEN表达上调。进一步检测AKT/mTOR信号相关蛋白的表达及磷酸化,显示Ataxin-3过表达时AKT、mTOR无明显变化,但AKT、mTOR磷酸化明显上调,真核翻译起始因子4E结合蛋白1(4E-BP1)磷酸化明显下降;而Ataxin-3敲除后,p-AKT、p-mTOR表达明显抑制,4E-BP1磷酸化明显上调。结论Ataxin-3在TC组织中呈高表达,且与TC进展有关,其可能机制为Ataxin-3抑制抗癌基因PTEN通过激活AKT/mTOR信号通路而促进TC细胞增殖。Objective To investigate the expression of spinocerebellar ataxia associated protein(Ataxin)-3 in testicular cancer(TC)and the mechanism by which Ataxin-3 promotes proliferation of TC cells.Methods A total of 38 paired specimens of TC tissues and adjacent tissues from patients undergoing orchiectomy in the Department of Urology,Fuyang People’s Hospital between January 2015 and May 2019 were included in the study.Immunohistochemical staining and real-time PCR were used to detect the expression of Ataxin-3.The proliferation of TC cells was detected by CCK8 assay.Levels of phosphatase tensin homology deleted on chromosome ten(PTEN)and AKT/mTOR signaling pathway-related proteins after overexpression or knockout of Ataxin-3 were detected by Western blotting.Results The expression levels of Ataxin-3 protein and mRNA in TC tissues were significantly higher than those in adjacent tissues(P<0.05).The expression of Ataxin-3 was not related to the patient age and pathological type of TC(P>0.05),but was significantly correlated with clinical staging and metastasis(P<0.05).Overexpression of Ataxin-3 protein in TCam-2 cells could promote cell proliferation,while transfecting siRNA into TCam-2 cells and knockout of Ataxin-3 could inhibit cell proliferation and invasion.The expression of PTEN protein in TCam-2 cells was significantly inhibited when Ataxin-3 was overexpressed,and the expression of PTEN was up-regulated after Ataxin-3 knockout.Further detection of the expression and phosphorylation of AKT/mTOR signal-related proteins showed that there were no obvious changes in AKT or mTOR expression when Ataxin-3 was overexpressed,but the phosphorylation of AKT and mTOR was up-regulated,and the 4E-BP1 phosphorylation was significantly decreased.After knockout of Ataxin-3,the expression of p-AKT and p-mTOR was significantly inhibited,and 4E-BP1 phosphorylation was significantly up-regulated.Conclusion Ataxin-3 is highly expressed in TC tissues and is involved in the progression of TC.The mechanism may be related to Ataxin-3 inhi

关 键 词：睾丸癌 脊髓小脑共济失调症相关蛋白-3 张力蛋白同源基因 AKT/mTOR信号通路 

分 类 号：R74[医药卫生—神经病学与精神病学] R73[医药卫生—临床医学]