Bmi-1在大肠癌组织中的表达及意义
摘要
目的
检测Bmi-1基因在大肠癌组织和正常组织中表达水平的差异;探讨Bmi-1基因的表达与大肠癌病理学特征的相关性及其在大肠癌发生发展中的可能作用机制。
材料与方法
收集我院50例大肠癌组织标本,50例正常组织作为对照,正常对照组织为距肿瘤边缘5cm以上的癌旁正常组织。肿瘤标本切取后迅速分成两份,一份用10%甲醛固定液进行固定,行病理学检查,另一份迅速放入液氮罐中冷冻备用。利用实时荧光定量PCR(FQ-PCR)法检测标本中Bmi-1mRNA的表达水平,同时利用免疫组化法检测Bmi-1和P16蛋白的表达,并结合大肠癌的临床病理学资料,分析Bmi-1基因的表达水平与年龄、性别、肿瘤的大小、Dukes分期、淋巴结转移、远处转移、血清CEA、血清CA19-9、P53及P16蛋白等临床病理特征的关系。
结果
Bmi-1mRNA及蛋白在大肠癌组织中的表达明显高于正常对照组织(P < 0.05),大肠癌组织Bmi-1蛋白表达与P16蛋白表达呈负相关关系。Bmi-1蛋白的表达水平与肿瘤体积大小、Dukes分期、淋巴结转移、远处转移和P16蛋白的表达水平密切相关,其阳性表达率分别为90%、40%(P < 0.05);96.2%、41.7% (P < 0.05) ;96.2%、41.7% (P < 0.05) ;92.3%、62.2% (P< 0.05) ;85.7%、33.3% (P < 0.05),即随着肿瘤体积增大、临床分期的增加Bmi-1蛋白在癌细胞内的含量逐渐增高;有淋巴结转移、远处转移者Bmi-1蛋白的阳表达率明显高于无淋巴结转移、远处转移者;而Bmi-1蛋白的表达水平与患者性别、年龄、血清CEA、CA19-9、P53等无相关性。
结论
1.Bmi-1mRNA及蛋白在大肠癌组织中的表达明显高于肿瘤周围正常组织。
2.Bmi-1基因可能通过抑制P16蛋白的表达而在大肠癌的发生发展中起重要作用。
3.Bmi-1基因在大肠癌组织中的表达水平与大肠癌的临床分期成正相关关系。
4.Bmi-1基因的表达水平可作为判断大肠癌预后的指标之一。
Objective
The purpose of this study was to investigate the relationship between the expression of Bmi-1 in colorectal cancer and the clinicopathologic features, and to investigate the possible mechanism in the development of the colorectal cancer.
Materrials and METHODS
We collected 50 tissue samples of colorectal cancer as the treatment group and another 50 normal tissue samples as the control group. The samples of normal tissue were obtained at least 5 cm from the edge of adjacent tumor tissue. The tumor tissue samples were dissected into two parts, one of which immediately put into the fixative resolution of neutral formalin for subsequent pathological examination, another researved into a tank full of liquid nitrogen. The expression level of Bmi-1 mRNA in the specimens were detected by Fluorescence Quantitative PCR. The expression of Bmi-1 and P16 protein were determined by immunohistochemistry. Based upon the pathological results, we analyzed whether there existed correlation between the expression level of Bmi-1 protein and clinicopathological characteristics such as age, gender, tumor size, Dukes stage, lymphnode metastasis, distant metastasis, serum CEA, serum CA19-9, P53 protein and P16 protein.
RESULTS
Compared with normal tissues, the carcinoma tissues are significantly higher in terms of the expression level of Bmi-1 gene and protein, and lower of P16 protein. Pathological parameters, including tumor size, DukesE stage, lymph node metastasis, distant metastasis and the the expression level of P16 protein, are all significantly correlated with the expression level of Bmi-1. The expression level of Bmi-1 protein is higher in cases of greater tumor size or more advanced clinical stage, higher in cases with lymph node metastasis than those without, higher in cases with metastatic colorectal carcinoma than those without. The age, gender, serum CEA, CA19-9 and P53 has no correlation with the expression of Bmi-1.
CONCLUSION
1. The expression level of Bmi-1 gene and protein in the carcinoma tissues is significantly higher than that of normal tissues.
2. Bmi-1 gene might involve with the tumorogenesis of human colorectal carcinoma by suspressing the P16 proteins.
3. The expression level of Bmi-1 protein in the carcinoma tissue is positively correlated with the grade of tumor staging.
4. The expression level of Bmi-1 could be a prognostic factor for patients with colorectal carcinoma.
检测Bmi-1基因在大肠癌组织和正常组织中表达水平的差异;探讨Bmi-1基因的表达与大肠癌病理学特征的相关性及其在大肠癌发生发展中的可能作用机制。
材料与方法
收集我院50例大肠癌组织标本,50例正常组织作为对照,正常对照组织为距肿瘤边缘5cm以上的癌旁正常组织。肿瘤标本切取后迅速分成两份,一份用10%甲醛固定液进行固定,行病理学检查,另一份迅速放入液氮罐中冷冻备用。利用实时荧光定量PCR(FQ-PCR)法检测标本中Bmi-1mRNA的表达水平,同时利用免疫组化法检测Bmi-1和P16蛋白的表达,并结合大肠癌的临床病理学资料,分析Bmi-1基因的表达水平与年龄、性别、肿瘤的大小、Dukes分期、淋巴结转移、远处转移、血清CEA、血清CA19-9、P53及P16蛋白等临床病理特征的关系。
结果
Bmi-1mRNA及蛋白在大肠癌组织中的表达明显高于正常对照组织(P < 0.05),大肠癌组织Bmi-1蛋白表达与P16蛋白表达呈负相关关系。Bmi-1蛋白的表达水平与肿瘤体积大小、Dukes分期、淋巴结转移、远处转移和P16蛋白的表达水平密切相关,其阳性表达率分别为90%、40%(P < 0.05);96.2%、41.7% (P < 0.05) ;96.2%、41.7% (P < 0.05) ;92.3%、62.2% (P< 0.05) ;85.7%、33.3% (P < 0.05),即随着肿瘤体积增大、临床分期的增加Bmi-1蛋白在癌细胞内的含量逐渐增高;有淋巴结转移、远处转移者Bmi-1蛋白的阳表达率明显高于无淋巴结转移、远处转移者;而Bmi-1蛋白的表达水平与患者性别、年龄、血清CEA、CA19-9、P53等无相关性。
结论
1.Bmi-1mRNA及蛋白在大肠癌组织中的表达明显高于肿瘤周围正常组织。
2.Bmi-1基因可能通过抑制P16蛋白的表达而在大肠癌的发生发展中起重要作用。
3.Bmi-1基因在大肠癌组织中的表达水平与大肠癌的临床分期成正相关关系。
4.Bmi-1基因的表达水平可作为判断大肠癌预后的指标之一。
Objective
The purpose of this study was to investigate the relationship between the expression of Bmi-1 in colorectal cancer and the clinicopathologic features, and to investigate the possible mechanism in the development of the colorectal cancer.
Materrials and METHODS
We collected 50 tissue samples of colorectal cancer as the treatment group and another 50 normal tissue samples as the control group. The samples of normal tissue were obtained at least 5 cm from the edge of adjacent tumor tissue. The tumor tissue samples were dissected into two parts, one of which immediately put into the fixative resolution of neutral formalin for subsequent pathological examination, another researved into a tank full of liquid nitrogen. The expression level of Bmi-1 mRNA in the specimens were detected by Fluorescence Quantitative PCR. The expression of Bmi-1 and P16 protein were determined by immunohistochemistry. Based upon the pathological results, we analyzed whether there existed correlation between the expression level of Bmi-1 protein and clinicopathological characteristics such as age, gender, tumor size, Dukes stage, lymphnode metastasis, distant metastasis, serum CEA, serum CA19-9, P53 protein and P16 protein.
RESULTS
Compared with normal tissues, the carcinoma tissues are significantly higher in terms of the expression level of Bmi-1 gene and protein, and lower of P16 protein. Pathological parameters, including tumor size, DukesE stage, lymph node metastasis, distant metastasis and the the expression level of P16 protein, are all significantly correlated with the expression level of Bmi-1. The expression level of Bmi-1 protein is higher in cases of greater tumor size or more advanced clinical stage, higher in cases with lymph node metastasis than those without, higher in cases with metastatic colorectal carcinoma than those without. The age, gender, serum CEA, CA19-9 and P53 has no correlation with the expression of Bmi-1.
CONCLUSION
1. The expression level of Bmi-1 gene and protein in the carcinoma tissues is significantly higher than that of normal tissues.
2. Bmi-1 gene might involve with the tumorogenesis of human colorectal carcinoma by suspressing the P16 proteins.
3. The expression level of Bmi-1 protein in the carcinoma tissue is positively correlated with the grade of tumor staging.
4. The expression level of Bmi-1 could be a prognostic factor for patients with colorectal carcinoma.
引文
[1] Park IK, Qian D, Kiel M, et al. Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem cells[J]. Nature, 2003,423 (6937):302-305.
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[3] Kim JH, Yoon SY, Kim CN, et al. The Bmi-1 oncoprotein is overexpressed in human colorectal cancer and correlates with the reduced p16INK4a /p14ARF proteins [J]. Cancer Lett,2004,203(2):217-224.
[4] Vonlanthen S, Heighway J, Altermatt HJ, et al. The bmi-1 oncoprotein is differentially expressed in non-small cell lung cancer and correlates with INK4A-ARF locus expression[J]. Br J Cancer,2001,84(10):1372-1376.
[5]冯艳,宋立兵,郭宝红等.Bmi-1基因在乳腺癌组织中的表达及意义[J].癌症,2007,26(2);154-157.
[6] Bea S, Tort F, PinyolM, et al. Bmi-1 gene amp lification and overexpression in hematological malignancies occur mainly in mantle celllymphomas[J]. Cancer Res,2001,61 (6):2409-2412.
[7] Kemenade FJ, van Raaphorst FM, Blokzijl T, et al. Coexp ression of Bmi-1 and EZH2 polycomb group p rotein is associated with cycling cells and degree ofmalignancy in B2cell non2Hodgkin lymphoma[J]. Blood,2001,97(12):3896 -3901.
[8]马国光,郭坤元,尚振川等.Bmi-1基因及其编码蛋白在SKM-1细胞系中的表达[J].肿瘤防治研究,2006,33(2):83-85.
[9] Sorrentino Brian P, et al. Clinical strategies for expansion of haematopoietic stem cells[J]. Nat Rev Immunol,2004,4(11):878-888.
[10] Silva J, Garcia JM, Pe?a C, et al. Implication of polycomb members Bmi-1, Mel-18 and Hpc-2 in the regulation of p16INK4a, p14ARF, h-TERT, and c-Myc expression in primary breast carcinomas[J]. Clinical Cancer Research,2006,12(23):6929-6936.
[11] Dimri GP, Martinez JL, Jacobs JJ, et al. The Bmi-1 oncogene induces telomerase activity and immortalizes human mammary epi-thelial cells[J]. Cancer Res,2002,62 (16):4736-4745.
[12]蔡刚,李闻捷,沈茵等.实时逆转录聚合酶链反应绝对定量试验优化的研究[J].上海医学检验杂志,2003,18(6):343-346.
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[22]李建平,谢明等.抑癌基因p16突变与乳腺癌的相关研究[J].2007,9(4):123-126.
[23] Jares P, Fernanclez Pl, Campo E, et a1. PRAD-1/cyclin D1 gene amplification correlates with messenger RNA over expression and tumor progression in human laryngeal carcinomas[J]. Cancers,1994,549(17):4813-4817.
[1]冯艳,宋立兵等.Bmi-1基因在乳腺癌组织中的表达及意[J]. 2007,26(2): 154-157.
[2] Kim RH, Kang MK, Shin KH, et al. Bmi-1 cooperates with human papillomavirus type 16 E6 to immortalize normal human oral keratinocytes[J]. Exp Cell Res,2007, 313 (3):462-472.
[3] Dimri GP, Martinez JL, Jacobs J J, et al. The BMI-1 oncogene induces telomerase activity and immortalizes human mammary epi-thelial cells[J]. Cancer Res,2002,62 (16):4736-4745.
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[6] Kim JH, Yoon SY, Kim CN, et al. The Bmi-1 oncoprotein is overexpressed in human colorectal cancer andcorrelates with the reduced p16INK4a /p14ARF proteins [J]. Cancer Lett,2004,203(2):217-224.
[7] Tateishik, Ohta M, Kanai F, et al. Dysregulated expression of stem cell factor Bmi-1 in precancerous lesions of the gastrointestinal tract[J]. Clinical Cancer Reseach, 2006, 12(23):6960-6966.
[8] Vonlanthen, S Heighway, J Altermatt HJ, etal. The Bmi-1 oncoprotein is differentially expressed in non-small cell lung cancer and correlates with INK4A-ARF locus expression[J]. British Journal Cancer 2001,84(10),1372-1376.
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[21] Jares P, Fernanclez Pl, Campo E, et a1. PRAD-1/cyclin D1 gene amplification correlates with messenger RNA over expression and tumor progression in humanlaryngeal carcinomas[J]. Cancers,1994,549(17):4813-4817.
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[25] T Wolfel, M Hauer, J Schneider, et a1. A p16INK4a-insensitive CDK mutant targeted by cytolytic T lymphocytes in a human melanoma[J]. Science, 1995, 269 (5228):1281-1284.
[26]王丽琼等.周期素D1与p16蛋白及Rb基因在大肠癌中的表达及意义[J].云南医药,2007,5(28):434-443.
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[28] Kim J H, Yoon SY, Kim CN, et al. The Bmi-1 oncoprotein is overexpressed in human colorectal cancer and correlates with the reduced p16IN K4a/ p14ARF proteins[J]. Cancer Lett,2004,203(2):217-224.
[2] Huang KH, Liu JH, Li XX, et al. Association of Bmi-1 mRNA expression with differentiation, metastasis and prognosis of gastric carcinoma[J]. Nan Fang Yi Ke Da Xue Xue Bao,2007,27(7):973-975.
[3] Kim JH, Yoon SY, Kim CN, et al. The Bmi-1 oncoprotein is overexpressed in human colorectal cancer and correlates with the reduced p16INK4a /p14ARF proteins [J]. Cancer Lett,2004,203(2):217-224.
[4] Vonlanthen S, Heighway J, Altermatt HJ, et al. The bmi-1 oncoprotein is differentially expressed in non-small cell lung cancer and correlates with INK4A-ARF locus expression[J]. Br J Cancer,2001,84(10):1372-1376.
[5]冯艳,宋立兵,郭宝红等.Bmi-1基因在乳腺癌组织中的表达及意义[J].癌症,2007,26(2);154-157.
[6] Bea S, Tort F, PinyolM, et al. Bmi-1 gene amp lification and overexpression in hematological malignancies occur mainly in mantle celllymphomas[J]. Cancer Res,2001,61 (6):2409-2412.
[7] Kemenade FJ, van Raaphorst FM, Blokzijl T, et al. Coexp ression of Bmi-1 and EZH2 polycomb group p rotein is associated with cycling cells and degree ofmalignancy in B2cell non2Hodgkin lymphoma[J]. Blood,2001,97(12):3896 -3901.
[8]马国光,郭坤元,尚振川等.Bmi-1基因及其编码蛋白在SKM-1细胞系中的表达[J].肿瘤防治研究,2006,33(2):83-85.
[9] Sorrentino Brian P, et al. Clinical strategies for expansion of haematopoietic stem cells[J]. Nat Rev Immunol,2004,4(11):878-888.
[10] Silva J, Garcia JM, Pe?a C, et al. Implication of polycomb members Bmi-1, Mel-18 and Hpc-2 in the regulation of p16INK4a, p14ARF, h-TERT, and c-Myc expression in primary breast carcinomas[J]. Clinical Cancer Research,2006,12(23):6929-6936.
[11] Dimri GP, Martinez JL, Jacobs JJ, et al. The Bmi-1 oncogene induces telomerase activity and immortalizes human mammary epi-thelial cells[J]. Cancer Res,2002,62 (16):4736-4745.
[12]蔡刚,李闻捷,沈茵等.实时逆转录聚合酶链反应绝对定量试验优化的研究[J].上海医学检验杂志,2003,18(6):343-346.
[13] Kamb A, Gruis NA, W eavevFeldhaus J, et a1. A cell cycle regulator potentially involved in genesis of many tumor types[J]. Science,l994, 264(5167):436-440.
[14] Shapiro GI, Edwards CD,Ewen ME, et a1. pl6INK4A participates in a G1 arrest checkpoint in response to DNA damage [J]. Mol Cell biod 1998,18(1):378-387.
[15]李波,邹扬等.循环核酸Bmi-1 mRNA在大肠癌中的检测及意义[J].苏州大学毕业论文.
[16] Tateishik, Ohta M, Kanai F, et al. Dysregulated expression of stem cell factor Bmi-1 in precancerous lesions of the gastrointestinal tract[J]. Clinical Cancer Reseach, 2006, 12(23):6960-6966.
[17] D Beach,T Wolfel, M Hauer, et a1. A p16INK4a-insensitive CDK4 mutant targeted by cytolytic T lymphocytes in a human melanoma[J]. Science , 1995, 269(5228): 1281-1284.
[18]王丽琼,张曦,许瑞吉等.周期素D1与p16蛋白及Rb基因在大肠癌中的表达及意义[J].云南医药2007,5(28):434-437.
[19] Song LB, Zeng MS, Liao WT, et al. Bmi-1 is a novel molecular marker of nasopharyngeal carcinoma progression and immortalizes primary human nasophary -ngeal epithelial cells[J]. Cancer Res,2006,66(12):6225-6232.
[20] Wang H, Pan K, Zhang HK, et al. Increased poly-comb-group oncongene Bmi-1 expression correlates with poor prognosis in hepatocellular carcinoma[J]. Cancer Res Clin Oncol,2008,134(5):535-541.
[21] Herman JG, Abouezzi Z, Borgen P1, et al. Inactivation of the CDKN2/Pl6/MTSl gene is frequently associated with abetrant DNA methytation all common human cancers[J]. Cancers,1995,55(1 8):4525-4530.
[22]李建平,谢明等.抑癌基因p16突变与乳腺癌的相关研究[J].2007,9(4):123-126.
[23] Jares P, Fernanclez Pl, Campo E, et a1. PRAD-1/cyclin D1 gene amplification correlates with messenger RNA over expression and tumor progression in human laryngeal carcinomas[J]. Cancers,1994,549(17):4813-4817.
[1]冯艳,宋立兵等.Bmi-1基因在乳腺癌组织中的表达及意[J]. 2007,26(2): 154-157.
[2] Kim RH, Kang MK, Shin KH, et al. Bmi-1 cooperates with human papillomavirus type 16 E6 to immortalize normal human oral keratinocytes[J]. Exp Cell Res,2007, 313 (3):462-472.
[3] Dimri GP, Martinez JL, Jacobs J J, et al. The BMI-1 oncogene induces telomerase activity and immortalizes human mammary epi-thelial cells[J]. Cancer Res,2002,62 (16):4736-4745.
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