Bmi-1、OPN在宫颈鳞癌中的表达及其与浸润转移的关系

发布时间：2018-04-28 06:32

本文选题：Bmi-1 + OPN　；参考：《郑州大学》2014年硕士论文

【摘要】：背景与目的宫颈癌是威胁女性健康最常见的第二大恶性肿瘤，且发病年龄逐渐年轻化，而影响其病死率及预后的主要因素是宫颈癌的浸润和转移。在宫颈癌的发展过程中，存在着各种与浸润转移相关的癌基因的激活和抑癌基因的失活。因此，临床中对宫颈癌的治疗和预后判断有赖于对宫颈癌的发生发展、浸润转移机制的分子生物学的研究。近年研究表明，上皮-间质转化(epithelial-mesenchymaltransformation，EMT)与上皮细胞来源的肿瘤的浸润、转移有关，在口腔癌、肺癌、乳腺癌、肝癌、结肠癌、卵巢癌、前列腺癌等许多恶性肿瘤的原位浸润和远处转移过程中发挥了重要作用[1]。E-钙粘蛋白（E-cadherin）的表达下调和波形蛋白（Vimentin）的表达上调是EMT发生的主要特征。原癌基因Bmi-1(B-cell-specific Moloneymurine leukemia virus insertion site l，，BMI-1)是多梳基因(polycomb group genes，PcG)家族中的成员之一，在细胞的生长、增殖、凋亡调节和干细胞的自我更新中起着非常重要的作用[2]。研究表明，Bmi-1在许多肿瘤中表达上调，与恶性肿瘤的恶性转化有明显相关性。骨桥蛋白（osteopontin，OPN）是一种钙结合磷酸蛋白。多项研究显示，骨桥蛋白与肿瘤的浸润、转移和恶性转化有密切相关性，所以OPN基因又被称为肿瘤转移相关基因。目前，在宫颈癌的浸润转移方面针对Bmi-1、OPN的研究文献较少。本课题通过检测Bmi-1、OPN和EMT分子标记蛋白E-cadherin、Vimentin在宫颈癌变过程中的表达，并分析其相关性，探讨Bmi-1、OPN在宫颈鳞癌浸润、转移过程中的作用。材料与方法 1、标本来源：收集2012年6月至2013年7月郑州大学第二附属医院病理科存档的宫颈蜡块标本126例，其中包括因子宫肌瘤行子宫全切术的正常宫颈组织20例、60例CIN组织（CINⅠ25例、CINⅡ19例和CINⅢ16例）和术前未经放化疗且术后病理确诊的宫颈鳞癌组织46例。 2、实验方法：采用免疫组化SP法检测20例正常宫颈组织、60例CIN组织（CINⅠ25例、CINⅡ19例和CINⅢ16例）和46例宫颈鳞癌组织中Bmi-1、OPN、 E-cadherin和Vimentin的表达，并分析其相关性。 3、统计方法：采用SPSS17.0统计软件包对数据进行分析处理，定性资料的比较采用卡方检验或Fisher确切概率法，利用2×2配对资料进行相关分析。以P0.05定义为差别有统计学意义。结果 1、Bmi-1的表达：正常宫颈组织、CIN和宫颈鳞状细胞癌组织中Bmi-1蛋白主要定位在细胞核中。Bmi-1在正常宫颈组、CIN组和宫颈鳞癌组中的表达逐渐升高，总体有差异（P＜0.05）；其中正常宫颈组和宫颈鳞癌组、CIN组和宫颈鳞癌组相比均有差异（P＜0.05）。Bmi-1在CINⅠ、CINⅡ和CINⅢ中的表达有差异（P＜0.05），其中CINⅠ和CINⅢ相比有差异（P＜0.05）。宫颈鳞癌组织中，Bmi-1蛋白的表达与患者的临床分期、组织学分级和淋巴结转移有关（P＜0.05）。 2、OPN的表达：正常宫颈组织、CIN和宫颈鳞状细胞癌组织中OPN蛋白主要定位在细胞质和细胞核中。OPN在正常宫颈组、CIN组和宫颈鳞癌组中的表达逐渐升高，总体有差异（P＜0.05）；其中正常宫颈组和宫颈鳞癌组、CIN组和宫颈鳞癌组比较均有差异（P＜0.05）。OPN在CINⅠ、CINⅡ和CINⅢ中的表达无差异（P＜0.05），其中两两比较也无差异（P＞0.05）。宫颈鳞癌组织中，OPN蛋白的表达与患者的临床分期、组织学分级、组织浸润深度和淋巴结转移有关（P＜0.05）。 3、E-cadherin的表达：在正常宫颈组织、CIN和宫颈鳞癌组织中E-cadherin蛋白主要定位于细胞膜。E-cadherin在正常宫颈组、CIN组和宫颈鳞癌组中的表达逐步降低，总体有差异（P＜0.05）；其中两两之间相比均有差异（P＜0.05）。E-cadherin在CINⅠ、CINⅡ和CINⅢ中的表达无差异（P＜0.05），其中两两比较差异也均无差异（P＞0.05）。宫颈鳞癌组织中，E-cadherin蛋白的表达与患者的临床分期、组织学分级、组织浸润深度和淋巴结转移有关（P＜0.05）。 4、Vimentin的表达：Vimentin在正常宫颈组织中不表达，在CIN和宫颈鳞状细胞癌组织中Vimentin蛋白主要定位在细胞质中。Vimentin在CIN组和宫颈鳞癌组中的表达逐步升高，总体有差异（P＜0.05）；其中正常宫颈组和宫颈鳞癌组、CIN组和宫颈鳞癌组相比均有差异（P＜0.05）。Vimentin在CINⅠ、CINⅡ和CINⅢ中的表达差异无统计学意义（P＜0.05），其中两两相比差别也均无统计学意义（P＞0.05）。宫颈鳞癌组织中，Vimentin蛋白的表达与患者的组织浸润深度、临床分期和淋巴结转移有关（P＜0.05）。 5、宫颈鳞癌组织中Bmi-1、OPN、E-cadherin以及Vimentin的相关性： ⑴Bmi-1与E-cadherin的表达是负相关（r=-0.531,P＜0.05），与Vimentin的表达是正相关（r=0.344,P＜0.05）； ⑵OPN与E-cadherin表达是负相关（r=-0.544,P＜0.05），与Vimentin表达是正相关（r=0.368,P＜0.05）； ⑶Bmi-1与OPN表达是正相关（r=0.507,P＜0.05）； ⑷E-cadherin与Vimentin表达是负相关（r=-0.341,P＜0.05）。结论 1、原癌基因Bmi-1、肿瘤转移相关基因OPN的高表达与宫颈鳞癌的组织学分级、临床分期和淋巴结转移密切相关，提示Bmi-1、OPN可能在宫颈鳞癌的发生发展和浸润转移过程当中发挥了重要作用。 2、Bmi-1、OPN可能是通过上皮-间质转化促进宫颈鳞癌的浸润和转移。 3、Bmi-1与OPN在宫颈鳞癌中的表达呈正相关，提示两者可能协同促进宫颈癌的浸润和转移。
[Abstract]:Background and purpose
Cervical cancer is the second most common malignant tumor that threatens the health of women, and the age of the disease is gradually younger. The main factor affecting the mortality and prognosis is the invasion and metastasis of cervical cancer. In the development of cervical cancer, there are various activation and inactivation of tumor suppressor genes associated with infiltration and metastasis. The treatment and prognosis of cervical cancer depends on the development of cervical cancer and the molecular biology of the mechanism of invasion and metastasis. Recent studies have shown that epithelial mesenchymal transformation (epithelial-mesenchymaltransformation, EMT) is related to the invasion and metastasis of epithelial cells, and in oral cancer, lung cancer, breast cancer, liver cancer, colon, and colon In situ infiltration and distant metastasis of many malignant tumors such as cancer, ovarian cancer and prostate cancer have played an important role in the downregulation of [1].E- cadherin (E-cadherin) expression and the up-regulation of vimentin (Vimentin) expression is the main feature of EMT. The proto oncogene Bmi-1 (B-cell-specific Moloneymurine leukemia virus insertion site L) BMI-1) is one of the members of the Polycomb group genes (PcG) family, which plays a very important role in cell growth, proliferation, apoptosis regulation and the self-renewal of stem cells. [2]. studies show that Bmi-1 is up regulated in many tumors and has a significant correlation with malignant transformation of malignant tumor. Osteopontin (osteopontin, OPN) It is a calcium binding phosphate protein. A number of studies have shown that osteopontin is closely related to tumor invasion, metastasis and malignant transformation, so the OPN gene is also known as tumor metastasis related genes. At present, there are few studies on Bmi-1 and OPN in the invasion and metastasis of cervical cancer. This topic is based on the detection of Bmi-1, OPN and EMT molecular markers. The expression of protein E-cadherin and Vimentin in cervical carcinogenesis was analyzed, and the correlation between Bmi-1 and OPN in the process of invasion and metastasis of cervical squamous cell carcinoma was discussed.
Materials and methods
1, specimen sources: 126 cases of cervix paraffin specimens collected from the pathology department of the Second Affiliated Hospital of Zhengzhou University from June 2012 to July 2013 were collected, including 20 cases of normal cervical tissue with hysteromyoma undergoing hysterectomy, 60 cases of CIN tissue (CIN I 25 cases, CIN II 19 cases and CIN III 16 cases), and preoperative and postoperative pathologic diagnosis of the uterus. 46 cases of squamous cell carcinoma of the neck.
2, experimental methods: the expression of Bmi-1, OPN, E-cadherin and Vimentin in 20 cases of normal cervical tissue, 60 cases of CIN tissue (CIN I 25 cases, CIN II 19 cases and CIN III 16 cases) and 46 cases of cervical squamous cell carcinoma were detected by immunohistochemical SP method, and the correlation was analyzed.
3, statistical method: the data were analyzed with SPSS17.0 software package. Qualitative data were compared with chi square test or Fisher exact probability method, and 2 x 2 paired data were used for correlation analysis. The difference was statistically significant with the definition of P0.05.
Result
1, Bmi-1 expression: the expression of Bmi-1 protein in normal cervical tissue, CIN and cervical squamous cell carcinoma was mainly located in the nucleus.Bmi-1 in the normal cervix group, the CIN group and the cervical squamous cell carcinoma group gradually increased (P < 0.05), and the normal cervix group and the cervical squamous cell carcinoma group, the CIN group and the cervical squamous cell carcinoma group were different (P). The expression of.Bmi-1 in CIN I, CIN II and CIN III was different (P < 0.05), and there was a difference between CIN I and CIN III (P < 0.05). The expression of Bmi-1 protein in cervical squamous cell carcinoma was related to the clinical stage, histological grade and lymph node metastasis (P < 0.05).
2, OPN expression: the expression of OPN protein in normal cervical tissue, CIN and cervical squamous cell carcinoma is mainly located in the cytoplasm and nucleus of.OPN in the normal cervix group, in the CIN group and in the cervical squamous cell carcinoma group, the overall difference is (P < 0.05), and the normal cervix group and the cervical squamous cell carcinoma group, the CIN group and the cervical squamous cell carcinoma group have a poor comparison. The expression of different (P < 0.05).OPN in CIN I, CIN II and CIN III was not different (P < 0.05), and there was no difference between 22 (P > 0.05). The expression of OPN protein in cervical squamous cell carcinoma was related to the clinical stage, histological grade, depth of tissue infiltration and lymph node metastasis (P < 0.05).
3, E-cadherin expression: the expression of E-cadherin protein in normal cervical tissue, CIN and cervical squamous cell carcinoma was mainly located in the cell membrane.E-cadherin in the normal cervix group, the CIN group and the cervical squamous cell carcinoma group gradually decreased (P < 0.05), and the ratio of 22 (P < 0.05) was different (P < 0.05).E-cadherin in CIN I, CIN II and CIN. There was no difference in expression in III (P < 0.05), and there was no difference in 22 of the differences (P > 0.05). In cervical squamous cell carcinoma, the expression of E-cadherin protein was related to the clinical stage, histological grade, depth of tissue infiltration and lymph node metastasis (P < 0.05).
4, Vimentin expression: Vimentin was not expressed in normal cervical tissues. The expression of Vimentin protein in the cytoplasm of CIN and cervical squamous cell carcinoma increased gradually in the cytoplasm and in the group of CIN and cervical squamous cell carcinoma (P < 0.05); in the normal cervix group and the cervical squamous cell carcinoma group, the CIN group and the cervical squamous cell carcinoma group. There was no significant difference in the expression of.Vimentin (P < 0.05) in CIN I, CIN II and CIN III (P < 0.05), and there was no statistically significant difference between 22 of them (P > 0.05). The expression of Vimentin protein in cervical squamous cell carcinoma was related to the depth of tissue infiltration, clinical stage and lymph node metastasis (P < 0.05).
5, the correlation of Bmi-1, OPN, E-cadherin and Vimentin in cervical squamous cell carcinoma:
The expression of Bmi-1 was negatively correlated with E-cadherin expression (r=-0.531, P < 0.05), and was positively correlated with the expression of Vimentin (r=0.344, P < 0.05).
(2) OPN and E-cadherin expression were negatively correlated (r=-0.544, P < 0.05), and positively correlated with Vimentin expression (r=0.368, P < 0.05).
There was a positive correlation between Bmi-1 and OPN expression (r=0.507, P < 0.05).
There was a negative correlation between E-cadherin and Vimentin expression (r=-0.341, P < 0.05).
conclusion
1, the high expression of proto oncogene Bmi-1 and tumor metastasis related gene OPN is closely related to the histological grade of cervical squamous cell carcinoma, clinical staging and lymph node metastasis, suggesting that Bmi-1, OPN may play an important role in the development of cervical squamous cell carcinoma and the process of invasion and metastasis.
2, Bmi-1 and OPN may promote the invasion and metastasis of cervical squamous cell carcinoma through epithelial mesenchymal transition.
3, there is a positive correlation between Bmi-1 and OPN expression in cervical squamous cell carcinoma, suggesting that both may promote the invasion and metastasis of cervical cancer.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【参考文献】