干細胞再生因子Bmi-1及PCNA在惡性黑素瘤中的表達
發(fā)布時間：2018-07-14 21:55
【摘要】：背景 惡性黑素瘤(malignant melanoma, MM)是一種高度惡性的,起源于皮膚黑素細胞的腫瘤。它轉(zhuǎn)移早、死亡率高、手術(shù)后易復(fù)發(fā)、對化療藥物不敏感,且病因及機理不明。經(jīng)過數(shù)十年的研究,盡管惡黑病人的5年存活率及預(yù)后都有顯著提高,但其發(fā)病危險系數(shù)及總體死亡率卻逐年上升,尤其在成人原發(fā)腫瘤中增加速度更快。惡性黑素瘤可以分為皮膚原發(fā)性黑素瘤(primary melanoma)和轉(zhuǎn)移性黑素瘤(metastatic melanoma),也稱侵襲性黑素瘤。轉(zhuǎn)移性黑素瘤與皮膚原發(fā)性黑素瘤相比具有病程更短,治療效果更差的特點。 多梳基因家族(Polycomb group, PcG)由多種與細胞周期和增殖相關(guān)的轉(zhuǎn)錄抑制子組成,是一個染色質(zhì)表觀遺傳修飾因子家族,最初是作為同源盒基因(Hox)的抑制因子被發(fā)現(xiàn),其成員在參與造血、前后軸形成等生命過程中的主要功能是使其靶基因沉默。在維持干細胞的生理活性方面,PcG對部分分化和發(fā)育相關(guān)基因表達抑制；在基因的分化表達過程中,它們對不同靶基因進行阻遏或去阻遏化。 Bmi-1(B-cell specific moloney murine leukemia virus insertion site 1)是多梳基因家族(Polycomb group, PcG)中的一員,其具有很強的自我更新能力,作為一種干細胞再生因子已經(jīng)證實在多種腫瘤干細胞(tumor stem cell, TSC)中表達異常增高。Bmi-1是1991年在荷蘭癌癥中心小鼠淋巴瘤細胞中作為一種原癌基因被發(fā)現(xiàn)的。新近研究顯示Bmi-1做為干細胞再生因子在多種腫瘤的TSC中表達高于非TSC,而且將其敲除后,腫瘤的自我更新能力、腫瘤大小、細胞克隆能力,以及動物體內(nèi)實驗的移植成瘤性和傳代能力等均明顯下降。 增殖細胞核抗原(proliferating cell nuclear antigen, PCNA)是一種分子量為36 KD的酸性蛋白,首先由Miyachi等[1]在系統(tǒng)性紅斑狼瘡患者血清中發(fā)現(xiàn)并命名。PCNA是真核細胞DNA合成所必需的輔酶,具有協(xié)調(diào)DNA復(fù)制、調(diào)控細胞周期等多種生物學(xué)功能,在調(diào)節(jié)DNA合成和細胞增殖方面起著重要作用。增殖旺盛的細胞中PCNA呈強陽性表達,而在靜止期細胞中為陰性表達,且其合成量與細胞增殖狀態(tài)成正相關(guān),在細胞G1期開始升高,于S期達到峰值,M期降至最低點,與DNA的合成量曲線相一致。因此,可以通過監(jiān)測PCNA的量來間接反映局部細胞的增殖狀態(tài),是細胞增殖的理想標記物。目前,有關(guān)Bmi-1在MM中的表達及作用機制尚不清楚。國內(nèi)外有關(guān)Bmi-1在MM中表達及意義的報道也十分罕見。為此,本研究采用免疫組化方法,對36例MM(皮膚原發(fā)性黑素瘤26例,轉(zhuǎn)移性黑素瘤10例),30例皮內(nèi)痣,5例人黑素瘤A375細胞裸鼠移植瘤標本中干細胞再生因子Bmi-1、PCNA的表達進行檢測,以期探討B(tài)mi-1、PCNA與MM的臨床病理聯(lián)系。 目的 探討干細胞再生因子Bmi-1及PCNA在惡性黑素瘤中的表達及臨床病理意義。 材料與方法 (1)收集從鄭州大學(xué)第一附屬醫(yī)院病理中心2007年1月至2009年9月手術(shù)切除的標本,其中惡性黑素瘤36例(26例皮膚原發(fā)性黑素瘤及10例轉(zhuǎn)移性黑素瘤)、皮內(nèi)痣30例。5例人黑素瘤A375細胞裸鼠移植瘤標本來自河南省腫瘤病理重點實驗室。 (2)所有病理標本均由兩位臨床經(jīng)驗豐富的病理醫(yī)師確診。 (3)免疫組化S-P法檢測36例MM,30例皮內(nèi)痣,5例A375裸鼠移植瘤中Bmi-1及PCNA的表達。 (4)數(shù)據(jù)采用SPSS 16.0統(tǒng)計軟件對數(shù)據(jù)采用獨立多組二分類資料的卡方檢驗、四格表Fisher精確概率法進行統(tǒng)計學(xué)分析處理,2×2配對資料的相關(guān)性分析。檢驗水準均為a=0.05。 結(jié)果 1.Bmi-1的表達 Bmi-1蛋白在惡性黑素瘤、皮內(nèi)痣中的陽性率分別為61.11%、20.00%二者比較差異有統(tǒng)計學(xué)意義(X2=11.323,P--0.0010.05),A375細胞裸鼠移植瘤為100.00%；皮膚原發(fā)性黑素瘤和轉(zhuǎn)移性黑素瘤中的陽性率分別為57.31%、70.00%,二者比較差異無統(tǒng)計學(xué)意義(X2=0.460,P0.05)。 2. PCNA的表達 PCNA蛋白在惡性黑素瘤、皮內(nèi)痣中的陽性率分別為80.56%、10.00%,二者比較差異有統(tǒng)計學(xué)意義(X2=32.614,P0.001),A375細胞裸鼠移植瘤為100.00%；皮膚原發(fā)性黑素瘤和轉(zhuǎn)移性黑素瘤中的陽性率分別為73.07%、100.00%,差異無統(tǒng)計學(xué)意義(X2=2.236,P0.05) 3.Bmi-1與PCNA相關(guān)性分析 惡性黑素瘤中,Bmi-1與PCNA蛋白陽性率呈顯著相關(guān)(X2=6.62,P=0.01,r=0.40)。 4.Bmi-1陽性率與患者性別、年齡、病程的關(guān)系 在惡性黑素瘤組織中,Bmi-1表達陽性率的高低與患者性別、年齡、病程無顯著相關(guān)性。((r1=0.115,r2=0.108,r3=0.205,P均0.05)。 結(jié)論 1.人惡性黑素瘤及A375細胞裸鼠移植瘤中Bmi-1均高表達,提示Bmi-1可能介入了本病的發(fā)病過程。 2.惡性黑素瘤中,Bmi-1與PCNA蛋白陽性率呈正相關(guān),表明腫瘤組織中存在增生活躍細胞,可能與腫瘤干細胞的存在有關(guān)。 3.皮膚原發(fā)性黑素瘤和轉(zhuǎn)移性黑素瘤中Bmi-1、PCNA的陽性率差異無統(tǒng)計學(xué)意義,提示二者在原發(fā)性和轉(zhuǎn)移性黑素瘤中表達一致。
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.5
本文編號：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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.5
【參考文獻】
相關(guān)期刊論文 前4條
1 徐文博;張江安;于建斌;龐琳娜;曹瑞祥;甄希;;ABC轉(zhuǎn)運蛋白ABCG2在皮膚黑素瘤中的表達[J];中國皮膚性病學(xué)雜志;2010年05期
2 林妙霞;文卓夫;馮智英;;大腸癌中Bmi-1的表達及其臨床病理意義[J];中國病理生理雜志;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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