干细胞再生因子Bmi-1及PCNA在恶性黑素瘤中的表达
发布时间：2018-07-14 21:55
【摘要】：背景 恶性黑素瘤(malignant melanoma, MM)是一种高度恶性的,起源于皮肤黑素细胞的肿瘤。它转移早、死亡率高、手术后易复发、对化疗药物不敏感,且病因及机理不明。经过数十年的研究,尽管恶黑病人的5年存活率及预后都有显著提高,但其发病危险系数及总体死亡率却逐年上升,尤其在成人原发肿瘤中增加速度更快。恶性黑素瘤可以分为皮肤原发性黑素瘤(primary melanoma)和转移性黑素瘤(metastatic melanoma),也称侵袭性黑素瘤。转移性黑素瘤与皮肤原发性黑素瘤相比具有病程更短,治疗效果更差的特点。 多梳基因家族(Polycomb group, PcG)由多种与细胞周期和增殖相关的转录抑制子组成,是一个染色质表观遗传修饰因子家族,最初是作为同源盒基因(Hox)的抑制因子被发现,其成员在参与造血、前后轴形成等生命过程中的主要功能是使其靶基因沉默。在维持干细胞的生理活性方面,PcG对部分分化和发育相关基因表达抑制；在基因的分化表达过程中,它们对不同靶基因进行阻遏或去阻遏化。 Bmi-1(B-cell specific moloney murine leukemia virus insertion site 1)是多梳基因家族(Polycomb group, PcG)中的一员,其具有很强的自我更新能力,作为一种干细胞再生因子已经证实在多种肿瘤干细胞(tumor stem cell, TSC)中表达异常增高。Bmi-1是1991年在荷兰癌症中心小鼠淋巴瘤细胞中作为一种原癌基因被发现的。新近研究显示Bmi-1做为干细胞再生因子在多种肿瘤的TSC中表达高于非TSC,而且将其敲除后,肿瘤的自我更新能力、肿瘤大小、细胞克隆能力,以及动物体内实验的移植成瘤性和传代能力等均明显下降。 增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)是一种分子量为36 KD的酸性蛋白,首先由Miyachi等[1]在系统性红斑狼疮患者血清中发现并命名。PCNA是真核细胞DNA合成所必需的辅酶,具有协调DNA复制、调控细胞周期等多种生物学功能,在调节DNA合成和细胞增殖方面起着重要作用。增殖旺盛的细胞中PCNA呈强阳性表达,而在静止期细胞中为阴性表达,且其合成量与细胞增殖状态成正相关,在细胞G1期开始升高,于S期达到峰值,M期降至最低点,与DNA的合成量曲线相一致。因此,可以通过监测PCNA的量来间接反映局部细胞的增殖状态,是细胞增殖的理想标记物。目前,有关Bmi-1在MM中的表达及作用机制尚不清楚。国内外有关Bmi-1在MM中表达及意义的报道也十分罕见。为此,本研究采用免疫组化方法,对36例MM(皮肤原发性黑素瘤26例,转移性黑素瘤10例),30例皮内痣,5例人黑素瘤A375细胞裸鼠移植瘤标本中干细胞再生因子Bmi-1、PCNA的表达进行检测,以期探讨Bmi-1、PCNA与MM的临床病理联系。 目的 探讨干细胞再生因子Bmi-1及PCNA在恶性黑素瘤中的表达及临床病理意义。 材料与方法 (1)收集从郑州大学第一附属医院病理中心2007年1月至2009年9月手术切除的标本,其中恶性黑素瘤36例(26例皮肤原发性黑素瘤及10例转移性黑素瘤)、皮内痣30例。5例人黑素瘤A375细胞裸鼠移植瘤标本来自河南省肿瘤病理重点实验室。 (2)所有病理标本均由两位临床经验丰富的病理医师确诊。 (3)免疫组化S-P法检测36例MM,30例皮内痣,5例A375裸鼠移植瘤中Bmi-1及PCNA的表达。 (4)数据采用SPSS 16.0统计软件对数据采用独立多组二分类资料的卡方检验、四格表Fisher精确概率法进行统计学分析处理,2×2配对资料的相关性分析。检验水准均为a=0.05。 结果 1.Bmi-1的表达 Bmi-1蛋白在恶性黑素瘤、皮内痣中的阳性率分别为61.11%、20.00%二者比较差异有统计学意义(X2=11.323,P--0.0010.05),A375细胞裸鼠移植瘤为100.00%；皮肤原发性黑素瘤和转移性黑素瘤中的阳性率分别为57.31%、70.00%,二者比较差异无统计学意义(X2=0.460,P0.05)。 2. PCNA的表达 PCNA蛋白在恶性黑素瘤、皮内痣中的阳性率分别为80.56%、10.00%,二者比较差异有统计学意义(X2=32.614,P0.001),A375细胞裸鼠移植瘤为100.00%；皮肤原发性黑素瘤和转移性黑素瘤中的阳性率分别为73.07%、100.00%,差异无统计学意义(X2=2.236,P0.05) 3.Bmi-1与PCNA相关性分析 恶性黑素瘤中,Bmi-1与PCNA蛋白阳性率呈显著相关(X2=6.62,P=0.01,r=0.40)。 4.Bmi-1阳性率与患者性别、年龄、病程的关系 在恶性黑素瘤组织中,Bmi-1表达阳性率的高低与患者性别、年龄、病程无显著相关性。((r1=0.115,r2=0.108,r3=0.205,P均0.05)。 结论 1.人恶性黑素瘤及A375细胞裸鼠移植瘤中Bmi-1均高表达,提示Bmi-1可能介入了本病的发病过程。 2.恶性黑素瘤中,Bmi-1与PCNA蛋白阳性率呈正相关,表明肿瘤组织中存在增生活跃细胞,可能与肿瘤干细胞的存在有关。 3.皮肤原发性黑素瘤和转移性黑素瘤中Bmi-1、PCNA的阳性率差异无统计学意义,提示二者在原发性和转移性黑素瘤中表达一致。
[Abstract]:background
Malignant melanoma (MM) is a highly malignant tumor derived from melanocytes of the skin. It has an early metastasis, high mortality, easy to relapse after operation, insensitive to chemotherapeutic drugs, and the etiology and mechanism is unknown. After decades of study, the 5 year survival rate and prognosis of people with malignant melanasis have been significantly improved, but they are in danger. The risk factor and overall mortality rate are increasing year by year, especially in adult primary tumors. Malignant melanoma can be divided into primary skin melanoma (primary melanoma) and metastatic melanoma (metastatic melanoma), also known as invasive melanoma. The effect of treatment is worse.
The Polycomb group (PcG) is composed of a variety of transcriptional suppressors associated with cell cycle and proliferation. It is a chromatin epigenetic modifier family. It was originally found as a inhibitory factor of the homologous box gene (Hox). The main function of its members in the life process, such as hematopoiesis, the formation of the anterior and posterior axis, is the target of the target. Gene silencing. In maintaining the physiological activity of stem cells, PcG inhibits the expression of partially differentiated and developmental related genes; in the process of gene differentiation, they repression or depressor of different target genes.
Bmi-1 (B-cell specific Moloney murine leukemia virus insertion site 1) is a member of the multi comb gene family (Polycomb group, PcG). It has a strong self-renewal capacity. As a stem cell regeneration factor, it has been proved to be highly expressed in a variety of tumor stem cells in 1991 in Holland. Cancer center mouse lymphoma cells are found as a proto oncogene. Recent studies have shown that Bmi-1 is expressed as a stem cell regeneration factor in the TSC of a variety of tumors that is higher than non TSC, and after it is knocked out, the self renewal capacity of the tumor, the size of the tumor, the cell clone ability, and the xenografts and passages in animal experiments The ability and so on decreased obviously.
Proliferating cell nuclear antigen (PCNA) is an acid protein with a molecular weight of 36 KD. First, Miyachi and other [1] are found in the serum of patients with systemic lupus erythematosus and named.PCNA as a coenzyme necessary for the synthesis of eukaryotic cell DNA. It has many biological functions, such as coordinating DNA replication, regulating cell cycle and other biological functions. It plays an important role in regulating the synthesis of DNA and cell proliferation. The strong positive expression of PCNA in the proliferating cells is negative in the stationary phase cells, and its synthesis is positively correlated with the cell proliferation state. It begins to rise in the G1 phase of the cell and reaches the peak in the S phase, and the M phase falls to the lowest point, which is consistent with the DNA synthesis curve. The proliferation state of local cells can be indirectly reflected by the quantity of PCNA, which is an ideal marker for cell proliferation. At present, the expression and mechanism of Bmi-1 in MM are not clear. The reports about the expression and significance of Bmi-1 in MM are also rare. Therefore, 36 cases of MM (skin origin) are used in this study. 26 cases of hair melanoma, 10 cases of metastatic melanoma, 30 cases of intradermal nevus, and 5 cases of human melanoma A375 cells in nude mice, the expression of stem cell regenerative factor Bmi-1 and PCNA were detected in order to explore the clinicopathological relationship between Bmi-1, PCNA and MM.
objective
Objective to investigate the expression of stem cell regenerating factor Bmi-1 and PCNA in malignant melanoma and its clinicopathological significance.
Materials and methods
(1) collected specimens from the pathology center of the First Affiliated Hospital of Zhengzhou University from January 2007 to September 2009, including 36 cases of malignant melanoma (26 cases of skin primary melanoma and 10 cases of metastatic melanoma), 30 cases of intradermal nevus and.5 human melanoma A375 cell xenografts from the Key Laboratory of tumor pathology in Henan province.
(2) all pathological specimens were diagnosed by two clinically experienced pathologists.
(3) immunohistochemical S-P method was used to detect the expression of Bmi-1 and PCNA in 36 cases of MM, 30 cases of intradermal nevus and 5 cases of A375 nude mice transplanted tumor.
(4) the data used SPSS 16 statistical software on the data using independent multiple group of two classified data of chi square test, four lattice Fisher accurate probability method for statistical analysis, 2 x 2 paired data correlation analysis. The test level is a=0.05.
Result
Expression of 1.Bmi-1
The positive rates of Bmi-1 protein in malignant melanoma and intradermal nevus were 61.11%, 20% and two were statistically significant (X2=11.323, P--0.0010.05), A375 cells were 100% in nude mice, and 57.31% and 70% in skin primary melanoma and metastatic melanoma, respectively, and two were not statistically significant differences (X2=0 .460, P0.05).
The expression of 2. PCNA
The positive rates of PCNA protein in malignant melanoma and intradermal nevus were 80.56% and 10% respectively. The differences were statistically significant (X2=32.614, P0.001), and A375 cells in nude mice were 100%, and the positive rates of skin primary melanoma and metastatic melanoma were 73.07% and 100%, respectively, and there was no statistical difference (X2=2.236, P0.05).
Analysis of correlation between 3.Bmi-1 and PCNA
In malignant melanoma, the positive rate of Bmi-1 was significantly correlated with PCNA protein (X2=6.62, P=0.01, r=0.40).
Relationship between 4.Bmi-1 positive rate and sex, age and course of disease
In malignant melanoma tissues, there was no significant correlation between the positive rate of Bmi-1 expression and the sex, age and duration of the disease ((r1=0.115, r2=0.108, r3=0.205, P all 0.05).
conclusion
The high expression of Bmi-1 in 1. malignant melanoma and A375 cells transplanted in nude mice suggests that Bmi-1 may be involved in the pathogenesis of this disease.
2. in malignant melanoma, the positive rate of Bmi-1 is positively correlated with PCNA protein, indicating that there are proliferative and active cells in tumor tissue, which may be related to the existence of tumor stem cells.
There is no significant difference in the positive rate of Bmi-1 and PCNA in 3. primary and metastatic melanoma of the skin, suggesting that the two are in the same expression in primary and metastatic melanoma.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.5
本文编号：2123077
[Abstract]:background
Malignant melanoma (MM) is a highly malignant tumor derived from melanocytes of the skin. It has an early metastasis, high mortality, easy to relapse after operation, insensitive to chemotherapeutic drugs, and the etiology and mechanism is unknown. After decades of study, the 5 year survival rate and prognosis of people with malignant melanasis have been significantly improved, but they are in danger. The risk factor and overall mortality rate are increasing year by year, especially in adult primary tumors. Malignant melanoma can be divided into primary skin melanoma (primary melanoma) and metastatic melanoma (metastatic melanoma), also known as invasive melanoma. The effect of treatment is worse.
The Polycomb group (PcG) is composed of a variety of transcriptional suppressors associated with cell cycle and proliferation. It is a chromatin epigenetic modifier family. It was originally found as a inhibitory factor of the homologous box gene (Hox). The main function of its members in the life process, such as hematopoiesis, the formation of the anterior and posterior axis, is the target of the target. Gene silencing. In maintaining the physiological activity of stem cells, PcG inhibits the expression of partially differentiated and developmental related genes; in the process of gene differentiation, they repression or depressor of different target genes.
Bmi-1 (B-cell specific Moloney murine leukemia virus insertion site 1) is a member of the multi comb gene family (Polycomb group, PcG). It has a strong self-renewal capacity. As a stem cell regeneration factor, it has been proved to be highly expressed in a variety of tumor stem cells in 1991 in Holland. Cancer center mouse lymphoma cells are found as a proto oncogene. Recent studies have shown that Bmi-1 is expressed as a stem cell regeneration factor in the TSC of a variety of tumors that is higher than non TSC, and after it is knocked out, the self renewal capacity of the tumor, the size of the tumor, the cell clone ability, and the xenografts and passages in animal experiments The ability and so on decreased obviously.
Proliferating cell nuclear antigen (PCNA) is an acid protein with a molecular weight of 36 KD. First, Miyachi and other [1] are found in the serum of patients with systemic lupus erythematosus and named.PCNA as a coenzyme necessary for the synthesis of eukaryotic cell DNA. It has many biological functions, such as coordinating DNA replication, regulating cell cycle and other biological functions. It plays an important role in regulating the synthesis of DNA and cell proliferation. The strong positive expression of PCNA in the proliferating cells is negative in the stationary phase cells, and its synthesis is positively correlated with the cell proliferation state. It begins to rise in the G1 phase of the cell and reaches the peak in the S phase, and the M phase falls to the lowest point, which is consistent with the DNA synthesis curve. The proliferation state of local cells can be indirectly reflected by the quantity of PCNA, which is an ideal marker for cell proliferation. At present, the expression and mechanism of Bmi-1 in MM are not clear. The reports about the expression and significance of Bmi-1 in MM are also rare. Therefore, 36 cases of MM (skin origin) are used in this study. 26 cases of hair melanoma, 10 cases of metastatic melanoma, 30 cases of intradermal nevus, and 5 cases of human melanoma A375 cells in nude mice, the expression of stem cell regenerative factor Bmi-1 and PCNA were detected in order to explore the clinicopathological relationship between Bmi-1, PCNA and MM.
objective
Objective to investigate the expression of stem cell regenerating factor Bmi-1 and PCNA in malignant melanoma and its clinicopathological significance.
Materials and methods
(1) collected specimens from the pathology center of the First Affiliated Hospital of Zhengzhou University from January 2007 to September 2009, including 36 cases of malignant melanoma (26 cases of skin primary melanoma and 10 cases of metastatic melanoma), 30 cases of intradermal nevus and.5 human melanoma A375 cell xenografts from the Key Laboratory of tumor pathology in Henan province.
(2) all pathological specimens were diagnosed by two clinically experienced pathologists.
(3) immunohistochemical S-P method was used to detect the expression of Bmi-1 and PCNA in 36 cases of MM, 30 cases of intradermal nevus and 5 cases of A375 nude mice transplanted tumor.
(4) the data used SPSS 16 statistical software on the data using independent multiple group of two classified data of chi square test, four lattice Fisher accurate probability method for statistical analysis, 2 x 2 paired data correlation analysis. The test level is a=0.05.
Result
Expression of 1.Bmi-1
The positive rates of Bmi-1 protein in malignant melanoma and intradermal nevus were 61.11%, 20% and two were statistically significant (X2=11.323, P--0.0010.05), A375 cells were 100% in nude mice, and 57.31% and 70% in skin primary melanoma and metastatic melanoma, respectively, and two were not statistically significant differences (X2=0 .460, P0.05).
The expression of 2. PCNA
The positive rates of PCNA protein in malignant melanoma and intradermal nevus were 80.56% and 10% respectively. The differences were statistically significant (X2=32.614, P0.001), and A375 cells in nude mice were 100%, and the positive rates of skin primary melanoma and metastatic melanoma were 73.07% and 100%, respectively, and there was no statistical difference (X2=2.236, P0.05).
Analysis of correlation between 3.Bmi-1 and PCNA
In malignant melanoma, the positive rate of Bmi-1 was significantly correlated with PCNA protein (X2=6.62, P=0.01, r=0.40).
Relationship between 4.Bmi-1 positive rate and sex, age and course of disease
In malignant melanoma tissues, there was no significant correlation between the positive rate of Bmi-1 expression and the sex, age and duration of the disease ((r1=0.115, r2=0.108, r3=0.205, P all 0.05).
conclusion
The high expression of Bmi-1 in 1. malignant melanoma and A375 cells transplanted in nude mice suggests that Bmi-1 may be involved in the pathogenesis of this disease.
2. in malignant melanoma, the positive rate of Bmi-1 is positively correlated with PCNA protein, indicating that there are proliferative and active cells in tumor tissue, which may be related to the existence of tumor stem cells.
There is no significant difference in the positive rate of Bmi-1 and PCNA in 3. primary and metastatic melanoma of the skin, suggesting that the two are in the same expression in primary and metastatic melanoma.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.5
【参考文献】
相关期刊论文 前4条
1 徐文博;张江安;于建斌;庞琳娜;曹瑞祥;甄希;;ABC转运蛋白ABCG2在皮肤黑素瘤中的表达[J];中国皮肤性病学杂志;2010年05期
2 林妙霞;文卓夫;冯智英;;大肠癌中Bmi-1的表达及其临床病理意义[J];中国病理生理杂志;2009年03期
3 ;Laminin induces the expression of cytokeratin 19 in hepatocellular carcinoma cells growing in culture[J];World Journal of Gastroenterology;2003年05期
4 ;Expression of nitric oxide synthase in human gastric carcinoma and its relation to p53, PCNA[J];World Journal of Gastroenterology;2005年01期
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