目的：探讨溶酶体相关4次跨膜蛋白质B（lysosome-associated protein transmembrane 4 beta，LAPTM4B）基因多态性与肝癌易感性的关系．方法：应用病例对照研究方法，收集190例肝癌患者、190例慢性乙型肝炎患者、175例健康献血者全血，...目的：探讨溶酶体相关4次跨膜蛋白质B（lysosome-associated protein transmembrane 4 beta，LAPTM4B）基因多态性与肝癌易感性的关系．方法：应用病例对照研究方法，收集190例肝癌患者、190例慢性乙型肝炎患者、175例健康献血者全血，分离白细胞，提取基因组DNA，采用特异性引物PCR方法，扩增LAPTM4B第一外显子5′UTR内的部分序列，对三组人群进行分析研究．x^2检验分析肝癌组与对照组LAPTM4B的基因多态性和其他相关因素的相关性．结果：LAPTM4B等位基因在三组观察对象中的分布，^＊1和^＊2在健康对照组的频率分别是75．71％和24．29％，慢肝组73．16％和26、84％，肝癌组中66．84％和33.45％，三组比较等位基因分布频率有统计学意义，健康对照组与肝癌组比较有统计学意义（x^2=6．979，P=0．008）．LAPTM4B的基因型LAPTM4B1^＊1型、LAPTM4B^＊1／2混合型和LAPTM482^＊2型在肝癌组中的频率分别是37.9％、57.9％和4．2％、慢肝组50．5％、45．3％和4．2％、健康对照组56．6％、38.3％和5．1％，三组间三种基因型分布频率比较有统计学意义（x^2=14．854，P〈0．005）.结论：基因型^＊1／2和等位基因^＊2可能与肝癌的发生有关．展开更多
目的探讨溶酶体相关4次跨膜蛋白b(lysosomeassociated protein transmembran e-4b,L A P T M4B)基因-35及结缔组织生长因子(connective tissue growth factor,CTGF)蛋白在食管胃交界部腺癌组织中的表达及其意义.方法采用免疫组织化学染...目的探讨溶酶体相关4次跨膜蛋白b(lysosomeassociated protein transmembran e-4b,L A P T M4B)基因-35及结缔组织生长因子(connective tissue growth factor,CTGF)蛋白在食管胃交界部腺癌组织中的表达及其意义.方法采用免疫组织化学染色及蛋白印迹分析检测137例食管胃交界部腺癌组织中LAPTM4B-35及CTGF蛋白的表达情况,分析蛋白表达与患者临床病理参数的关系及对患者预后的影响.结果LAPTM4B-35蛋白在89.1%的食管胃交界部腺癌组织中呈高表达状态,LAPTM4B-35高表达组癌细胞发生远处转移(P=0.011)、肿瘤进展至TNMⅢ/Ⅳ期(P=0.026)显著多于低表达组.CTGF蛋白在51.1%的癌组织中呈高表达状态,CTGF高表达组发生远处转移(P=0.033)、肿瘤直径>5 cm(P=0.021)均多于低表达组.癌组织中LAPTM4B-35蛋白与CTGF蛋白表达之间呈正相关关系(r=0.218,P=0.010).单因素生存分析显示LAPTM4B-35、CTGF蛋白高表达的患者其术后生存时间分别显著低于LAPTM4B-35、CTGF蛋白低表达的患者(双侧log-rank检验,P<0.001).LAPTM4B-35蛋白表达(P<0.001)、发生远处转移(P<0.001)、淋巴结转移(P=0.007)、查见脉管癌栓(P=0.022)及患者性别(P<0.001)是影响食管胃交界部腺癌患者的独立预后因子.结论LAPTM4B-35及CTGF蛋白的高表达与肿瘤的侵袭转移及不良预后密切相关.展开更多
AIM: To produce high-quality polydonal antibody to lysosome associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellu...AIM: To produce high-quality polydonal antibody to lysosome associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC).METHODS: The 297 bp 5' end of LAPTM4BcDNA was obtained by PCR and inserted into prokaryotic expression vector pGEXKG. Then the recombinant pGEX-KG-N1-99 was transformedinto E. coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharoseTM 4B agarose. The purified GST-LAPTM4B-N1-99 was characterized by SDSPAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N1-99 was isolated from 1 L of bacteria. The GST-LAPTM4B-N1-99produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4B-N1-99-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely correlated with the differentiation of HCC.展开更多
文摘目的：探讨溶酶体相关4次跨膜蛋白质B（lysosome-associated protein transmembrane 4 beta，LAPTM4B）基因多态性与肝癌易感性的关系．方法：应用病例对照研究方法，收集190例肝癌患者、190例慢性乙型肝炎患者、175例健康献血者全血，分离白细胞，提取基因组DNA，采用特异性引物PCR方法，扩增LAPTM4B第一外显子5′UTR内的部分序列，对三组人群进行分析研究．x^2检验分析肝癌组与对照组LAPTM4B的基因多态性和其他相关因素的相关性．结果：LAPTM4B等位基因在三组观察对象中的分布，^＊1和^＊2在健康对照组的频率分别是75．71％和24．29％，慢肝组73．16％和26、84％，肝癌组中66．84％和33.45％，三组比较等位基因分布频率有统计学意义，健康对照组与肝癌组比较有统计学意义（x^2=6．979，P=0．008）．LAPTM4B的基因型LAPTM4B1^＊1型、LAPTM4B^＊1／2混合型和LAPTM482^＊2型在肝癌组中的频率分别是37.9％、57.9％和4．2％、慢肝组50．5％、45．3％和4．2％、健康对照组56．6％、38.3％和5．1％，三组间三种基因型分布频率比较有统计学意义（x^2=14．854，P〈0．005）.结论：基因型^＊1／2和等位基因^＊2可能与肝癌的发生有关．
文摘目的探讨溶酶体相关4次跨膜蛋白b(lysosomeassociated protein transmembran e-4b,L A P T M4B)基因-35及结缔组织生长因子(connective tissue growth factor,CTGF)蛋白在食管胃交界部腺癌组织中的表达及其意义.方法采用免疫组织化学染色及蛋白印迹分析检测137例食管胃交界部腺癌组织中LAPTM4B-35及CTGF蛋白的表达情况,分析蛋白表达与患者临床病理参数的关系及对患者预后的影响.结果LAPTM4B-35蛋白在89.1%的食管胃交界部腺癌组织中呈高表达状态,LAPTM4B-35高表达组癌细胞发生远处转移(P=0.011)、肿瘤进展至TNMⅢ/Ⅳ期(P=0.026)显著多于低表达组.CTGF蛋白在51.1%的癌组织中呈高表达状态,CTGF高表达组发生远处转移(P=0.033)、肿瘤直径>5 cm(P=0.021)均多于低表达组.癌组织中LAPTM4B-35蛋白与CTGF蛋白表达之间呈正相关关系(r=0.218,P=0.010).单因素生存分析显示LAPTM4B-35、CTGF蛋白高表达的患者其术后生存时间分别显著低于LAPTM4B-35、CTGF蛋白低表达的患者(双侧log-rank检验,P<0.001).LAPTM4B-35蛋白表达(P<0.001)、发生远处转移(P<0.001)、淋巴结转移(P=0.007)、查见脉管癌栓(P=0.022)及患者性别(P<0.001)是影响食管胃交界部腺癌患者的独立预后因子.结论LAPTM4B-35及CTGF蛋白的高表达与肿瘤的侵袭转移及不良预后密切相关.
基金Supported by the 248 Major R&D Program of Beijing, No. H020220020310, and Special Fund for Promotion of Education, Ministry of Education, China
文摘AIM: To produce high-quality polydonal antibody to lysosome associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC).METHODS: The 297 bp 5' end of LAPTM4BcDNA was obtained by PCR and inserted into prokaryotic expression vector pGEXKG. Then the recombinant pGEX-KG-N1-99 was transformedinto E. coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharoseTM 4B agarose. The purified GST-LAPTM4B-N1-99 was characterized by SDSPAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N1-99 was isolated from 1 L of bacteria. The GST-LAPTM4B-N1-99produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4B-N1-99-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely correlated with the differentiation of HCC.
