肿瘤干细胞标志物usp22在人膀胱移行细胞癌中的表达及临床意义
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摘要
目的研究泛素特异肽酶22(ubiquitin specific peptidase 22,USP22)在膀胱移行细胞癌组织及细胞中的基因及蛋白表达,并分析在膀胱癌组织和细胞中表达的差异情况,结合临床资料,探讨其可能的因果关系,进而得出相关性意义。
方法分析收集自2008年12月至2009年5月华中科技大学附属协和医院泌尿外科膀胱癌手术切除的肿瘤标本43例,其恶性度及病理分期都经过病理报告证实无误,正常膀胱组织31例,取材于癌旁远端的正常粘膜组织,其中男性患者30人,女性患者13人,年龄约在41-77岁之间,平均57.6岁,取材均是肉眼明显可分辨的癌组织,并远离肿瘤的切缘,防止因电刀高温引起的周围组织变性,患者在接受手术前均未接受放疗或者化疗。通过免疫组织化学法检测病例标本的usp22蛋白表达；及该院中心实验室保存低分化的EJ膀胱癌细胞,用Trizol抽提其RNA,蛋白裂解液获得蛋白,并和膀胱癌组织一起做检测,经逆转录及实时定量聚合酶链反应检测不同类型标本和细胞中usp22的表达；；观察不同类别标本的数据差异并与临床资料相结合,进行相关性分析；设计特异性siRNA通过转染沉默EJ细胞usp22基因,观察对照组和实验组的细胞生长传代增值情况后做出分析。
结果泛素特异肽酶22 mRNA在人正常膀胱组织中表达量很低,在膀胱移行细胞癌中表达明显升高,具有显著的差异性(p<0.01),在低度恶性的组织中其表达要低于中高度恶性组织中的表达(p<0.05).免疫组织化学显示膀胱移行细胞癌组织泛素特异肽酶22阳性率为87.5%(36/43),高于正常膀胱黏膜组织35.4%(11/31),P<0.05,且随着恶性度的增加,阳性率升高。在但是与性别年龄等因素无关(p>0.05)；活的膀胱癌细胞株中有同样类似的表达,恶性度较高的EJ细胞中mRNA表达也和3期膀胱癌细胞相似.
结论泛素特异肽酶22作为一种潜在的肿瘤干细胞标记物,可能上调了膀胱组织中某些基因的表达,进而引起正常细胞的恶变,在膀胱癌的发生和发展过程中有可能起着重要的作用；它在膀胱组织中的表达同恶性度或分级成正相关,有望成为判断病情发展的有用指标,同时很有可能成为该肿瘤早期诊断的重要指标,并成为研制抗肿瘤新药中的一个可用靶点。但其作用机制还需要进一步的研究来证实。
Objective:To investigate ubiquitin specific peptide enzyme 22 in bladder transitional cell carcinoma of the gene and tissue and cell protein expression, and analysis of bladder cancer in different malignant degrees in tissues and cells, and in combination with the express differences clinical information, discuss the possible causality, and a conclusion that come from the correlation significance
Methods:Analysis collected from December 2008 to 2009 May huazhong university of science and technology affiliated xiehe hospital uropoiesis surgical resection of the tumor specimen bladder surgery, the 43 cases were malignant degrees and pathologic stage through pathologic reports prove correct, normal bladder tissue, based on cancer in 31 cases of distant normal mucosa tissue by 30 people, including male patients, women were 13 people, around the age between 41-77 years old, average age 57.6 years, draw materials are obviously the naked eye can identify the cancerous tissue, and stay away from the tumor, prevent the cut edge caused by high temperature electrotome surrounding tissue degeneration, patients undergoing surgery were not accepting radiotherapy before or chemotherapy. According to its malignant degrees grading divided into four groups; And central lab save nays EJ and bladder BIU eighty-seven lines, extraction its RNA and protein, press extraction source different classification, then after a retrovirus and real-time quantitatie polymerase chain reaction testing different type specimens and cells usp22 expression; Through the test specimen immunohistochemistry methods were used usp22 protein expression; Observe different categories of data with differences and specimens, combined with clinical data relevant analysis.
Results:ubiquitin specific peptide enzyme22 in normal bladder tissue mRNA expressions is very low, quantity expressed in transitional cell carcinoma increased significantly, has the remarkable difference (p<0.01), in the organization of low-grade malignant lower than its expression in the organization highly malignant express (p<0.05). Immunohistochemistry display the positive rate in BTCC samples was 87.5%for 22 (36/43), higher than normal bladder mucous membrane tissues 35.4%(11/31), p<0.05, and with the increase of malignant degrees, positive elevated. But has nothing to do with factors such as age and gender (p>0.05). In the live bladder cancer cell lines, it have similar expression, EJ cells mRNA has higher expression biu-87 cells
Conclusion:ubiquitin specific peptide enzyme 22 as a potential cancer stem cells marker, may raise the expression of some bladder tissue gene, resulting in the malignant transformation, normal cells in the bladder cancer occurrence and development process may play an important role; The expression in bladder tissue with malignant degrees or classification, is expected to be a positive correlation judgment illness development, while useful indexes are most likely to become the early diagnosis of tumors, and become an important index of a developed antitumor drugs available targets. But further studies are needed to explain its mechanism
方法分析收集自2008年12月至2009年5月华中科技大学附属协和医院泌尿外科膀胱癌手术切除的肿瘤标本43例,其恶性度及病理分期都经过病理报告证实无误,正常膀胱组织31例,取材于癌旁远端的正常粘膜组织,其中男性患者30人,女性患者13人,年龄约在41-77岁之间,平均57.6岁,取材均是肉眼明显可分辨的癌组织,并远离肿瘤的切缘,防止因电刀高温引起的周围组织变性,患者在接受手术前均未接受放疗或者化疗。通过免疫组织化学法检测病例标本的usp22蛋白表达；及该院中心实验室保存低分化的EJ膀胱癌细胞,用Trizol抽提其RNA,蛋白裂解液获得蛋白,并和膀胱癌组织一起做检测,经逆转录及实时定量聚合酶链反应检测不同类型标本和细胞中usp22的表达；；观察不同类别标本的数据差异并与临床资料相结合,进行相关性分析；设计特异性siRNA通过转染沉默EJ细胞usp22基因,观察对照组和实验组的细胞生长传代增值情况后做出分析。
结果泛素特异肽酶22 mRNA在人正常膀胱组织中表达量很低,在膀胱移行细胞癌中表达明显升高,具有显著的差异性(p<0.01),在低度恶性的组织中其表达要低于中高度恶性组织中的表达(p<0.05).免疫组织化学显示膀胱移行细胞癌组织泛素特异肽酶22阳性率为87.5%(36/43),高于正常膀胱黏膜组织35.4%(11/31),P<0.05,且随着恶性度的增加,阳性率升高。在但是与性别年龄等因素无关(p>0.05)；活的膀胱癌细胞株中有同样类似的表达,恶性度较高的EJ细胞中mRNA表达也和3期膀胱癌细胞相似.
结论泛素特异肽酶22作为一种潜在的肿瘤干细胞标记物,可能上调了膀胱组织中某些基因的表达,进而引起正常细胞的恶变,在膀胱癌的发生和发展过程中有可能起着重要的作用；它在膀胱组织中的表达同恶性度或分级成正相关,有望成为判断病情发展的有用指标,同时很有可能成为该肿瘤早期诊断的重要指标,并成为研制抗肿瘤新药中的一个可用靶点。但其作用机制还需要进一步的研究来证实。
Objective:To investigate ubiquitin specific peptide enzyme 22 in bladder transitional cell carcinoma of the gene and tissue and cell protein expression, and analysis of bladder cancer in different malignant degrees in tissues and cells, and in combination with the express differences clinical information, discuss the possible causality, and a conclusion that come from the correlation significance
Methods:Analysis collected from December 2008 to 2009 May huazhong university of science and technology affiliated xiehe hospital uropoiesis surgical resection of the tumor specimen bladder surgery, the 43 cases were malignant degrees and pathologic stage through pathologic reports prove correct, normal bladder tissue, based on cancer in 31 cases of distant normal mucosa tissue by 30 people, including male patients, women were 13 people, around the age between 41-77 years old, average age 57.6 years, draw materials are obviously the naked eye can identify the cancerous tissue, and stay away from the tumor, prevent the cut edge caused by high temperature electrotome surrounding tissue degeneration, patients undergoing surgery were not accepting radiotherapy before or chemotherapy. According to its malignant degrees grading divided into four groups; And central lab save nays EJ and bladder BIU eighty-seven lines, extraction its RNA and protein, press extraction source different classification, then after a retrovirus and real-time quantitatie polymerase chain reaction testing different type specimens and cells usp22 expression; Through the test specimen immunohistochemistry methods were used usp22 protein expression; Observe different categories of data with differences and specimens, combined with clinical data relevant analysis.
Results:ubiquitin specific peptide enzyme22 in normal bladder tissue mRNA expressions is very low, quantity expressed in transitional cell carcinoma increased significantly, has the remarkable difference (p<0.01), in the organization of low-grade malignant lower than its expression in the organization highly malignant express (p<0.05). Immunohistochemistry display the positive rate in BTCC samples was 87.5%for 22 (36/43), higher than normal bladder mucous membrane tissues 35.4%(11/31), p<0.05, and with the increase of malignant degrees, positive elevated. But has nothing to do with factors such as age and gender (p>0.05). In the live bladder cancer cell lines, it have similar expression, EJ cells mRNA has higher expression biu-87 cells
Conclusion:ubiquitin specific peptide enzyme 22 as a potential cancer stem cells marker, may raise the expression of some bladder tissue gene, resulting in the malignant transformation, normal cells in the bladder cancer occurrence and development process may play an important role; The expression in bladder tissue with malignant degrees or classification, is expected to be a positive correlation judgment illness development, while useful indexes are most likely to become the early diagnosis of tumors, and become an important index of a developed antitumor drugs available targets. But further studies are needed to explain its mechanism
引文
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