丝裂原活化蛋白激酶和哺乳动物雷帕霉素靶蛋白激活剂3在膀胱癌中的表达及其临床意义

Expression and Clinical Significance of Late Endosomal/Lysosomal Adaptor, Mitogen-activated Protein Kinase and Mammalian Target of Rapamycin Activator 3 in Bladder Carcinoma

目的探讨丝裂原活化蛋白激酶和哺乳动物雷帕霉素靶蛋白激活剂3(LAMTOR3)在膀胱癌中的表达及其临床意义。方法检索Oncomine和Expression Atlas数据库,获得LAMTOR3相关的基因芯片数据,分析LAMTOR3在膀胱癌细胞及组织中的表达及其与膀胱癌临床病理特征的关系。采用RT-PCR、Western blot和免疫组织化学法检测LAMTOR3在膀胱癌细胞、癌组织及其配对的正常组织中的表达,验证其表达和临床相关性。结果Expression Atlas数据库检索结果显示,LAMTOR3在20株膀胱癌细胞中的基础表达水平存在差异,其中,在Hs 172.T、HT-1376、RT4、JMSU-1和T24细胞株中呈较高表达。Oncomine数据库检索结果显示,LAMTOR3在浸润性(t=2.857,P=0.005)及非浸润性膀胱癌组织(t=3.105,P=0.003)中的表达均明显高于正常膀胱组织,病理分级越高的膀胱癌组织中表达越高(P<0.05)。实验验证结果显示,LAMTOR3 mRNA在膀胱癌细胞株UMUC3(t=10.84,P=0.0084)、J82(t=21.75,P=0.0021)、5637(t=45.88,P=0.0005)和T24(t=87.58,P=0.0001)中的表达明显高于正常人膀胱永生化细胞SV-HUC-1,在膀胱癌组织中的表达也明显高于正常组织(P<0.05);LAMTOR3蛋白水平在癌组织中的表达也明显高于正常组织(P<0.05)。免疫组织化学检测结果显示,LAMTOR3蛋白在癌组织中高表达,在浸润性癌组织中较非浸润性癌组织高表达,其表达水平与膀胱癌临床分期(χ^2=9.189,P=0.002)、病理分级(χ^2=4.746,P=0.029)和淋巴结转移(χ^2=6.210,P=0.013)密切相关,但与性别(χ^2=0.965,P=0.326)、年龄(χ^2=2.126,P=0.145)、远处转移(χ^2=1.261,P=0.261)等无明显相关性。结论LAMTOR3在膀胱癌细胞及组织中高表达,与膀胱癌发生发展密切相关。

Objective To investigate the expression and clinical significance of late endosomal/lysosomal adaptor,mitogen-activated protein kinase and mammalian target of rapamycin activator 3(LAMTOR3)in bladder carcinoma. Methods Oncomine and Expression Atlas were used to extract the useful mining gene chip database for analyzing the expression of LAMTOR3 in bladder carcinoma tissues and cell lines,and the correlation of LAMTOR3 with the clinicopathological features were analyzed.RT-PCR,Western blot,and immunohistochemistry were performed to detect the expression of LAMTOR3 in bladder carcinoma cell lines,specimens,and adjacent normal tissues for verifying the results exploited from the above databases. Results The Expression Atlas showed that LAMTOR3 had high expressions in Hs172.T,HT-1376,RT4,JMSU-1,and T24 cell lines among 20 bladder carcinoma cell lines,among which the LAMTOR3 expression was different.Oncomine reported that LAMTOR3 expression in bladder carcinoma,including invasive( t=2.857, P=0.005)and non-invasive carcinoma( t=3.105, P=0.003),was significantly higher than that in adjacent normal tissues.The expression of LAMTOR3 was positively correlated with pathological grade( P <0.05).The expressions of LAMTOR3 mRNA in bladder carcinoma cell lines,including UMUC3( t=10.84, P=0.0084),J82( t=21.75, P=0.0021),5637( t=45.88, P=0.0005),and T24( t=87.58, P=0.0001) were significantly higher than that in normal bladder cell line SV-HUC-1,while its expression in bladder carcinoma tissues was significantly higher than that in adjacent normal tissues( P <0.05),so was its protein level in tissues( P <0.05). Immunohistochemistry showed that LAMTOR3 protein was over-expressed in bladder carcinoma tissues;its level in invasive carcinoma tissues was higher than that in no-invasive carcinoma tissues and was related closely with the clinical stages(χ^ 2=9.189, P=0.002),pathological grades(χ^ 2=4.746, P=0.029),and lymphatic metastasis(χ^2=6.210, P= 0.013)but had no significant correlation to sex(χ^2=0.965, P=0.326),age(χ^2=2.126, P=0.145),and distant metastasis(χ^2=1.261, P=0.261). Conclusion LAMTOR3 is highly expressed in bladder carcinoma cell lines and tissues and plays a key role in the development and progression of bladder carcinoma.