【作者】 杨晓；

【导师】 刘玉玲；

【作者基本信息】 郑州大学 ， 妇产科学， 2013， 硕士

【摘要】 背景与目的宫颈癌是仅次于乳腺癌之后,最常见的妇科恶性肿瘤之一,每年新发现的病例大多数集中在发展中国家,特别是贫困地区,近年来其发病有年轻化的趋势,以宫颈鳞状细胞浸润癌为主,总体预后不佳,对我国女性的健康和生命安全造成了严重威胁。高危型人乳头瘤病毒(HPV)持续感染是宫颈癌的主要病因,但并不是唯一因素,宫颈癌的发生发展是多因素、多阶段、多步骤共同作用的结果,因此需要进一步寻找和研究有临床意义的和宫颈癌相关的指标。近年研究发现转移相关基因1(metastasis-associated gene1,MTA1)是一个与肿瘤转移相关的基因；目前关于SLP-2(stomatin-like protein2)蛋白在多种肿瘤组织中的研究显示：与正常配对的多种肿瘤组织中,SLP-2蛋白的表达是上调的,而SLP-2在宫颈癌组织中的研究较少,在宫颈癌组织中SLP-2的表达是上调还是下调?与临床病理参数之间的关系如何?是否与MTA1具有相关性?就是本文研究的重点。材料与方法1、标本来源：收集2011年5月至2012年5月在郑州大学第二附属医院妇产科因子宫肌瘤行子宫全切术的正常宫颈组织48例(A组)、宫颈上皮内瘤变(cervical intraepithelial neoplasia CIN)患者的宫颈组织43例(B组)、术前未经放化疗且有明确病理诊断的宫颈鳞癌组织42例(C组),并采集病理科存档的蜡块标本。2、实验方法：采用免疫组化法检测不同宫颈组织中MTA1蛋白和SLP-2蛋白的表达及分布,RT-PCR法检测不同宫颈组织中MTA1mRNA和SLP-2mRNA的水平变化。3、统计方法：采用SPSS17.0统计软件包进行数据分析,定量资料的比较采用t检验和单因素方差分析,率的比较采用χ2检验,相关性分析采用2×2配对资料的关联性分析和pearson相关分析。检验水准a=0.05(双侧)。结果1、MTA1的表达情况：在正常宫颈组织MTA1蛋白主要定位于细胞核,CIN及宫颈鳞癌MTA1蛋白出现了细胞膜、细胞核和细胞浆多种形式的蛋白定位,MTA1蛋白在A组、B组及C组中的阳性表达呈上升趋势,MTA1mRNA在A组、B组及C组中的表达水平亦呈上升趋势,在A组中的表达显著低于B组(P<0.05)及C组(P<0.05),而B组中MTA1的表达低于C组,但无统计学意义(P>0.05)。MTA1蛋白和MTAlmRNA在宫颈鳞癌中的过表达与患者的年龄、临床分期、组织分化程度、肿瘤大小无关(P>0.05),而与患者的淋巴结转移、肿瘤浸润深度密切相关(P<0.05)。宫颈鳞癌患者有淋巴结转移和随着肿瘤浸润深度的加深,MTA1蛋白的过表达有升高的趋势。2、SLP-2的表达情况：在正常宫颈组织中SLP-2蛋白主要定位在细胞浆和细胞膜,CIN及宫颈鳞癌中SLP-2蛋白主要表现为细胞浆染色。SLP-2蛋白的阳性表达在A组、B组及C组中呈下降趋势,SLP-2mRNA的表达水平在A组、B组及C组中亦呈下降趋势,在C组中的表达明显低于A组(P<0.05)及B组(P<0.05),而B组中SLP-2的表达明显低于A组(P<0.05)。SLP-2的表达与宫颈鳞癌患者的年龄、临床分期、有无淋巴结转移、肿瘤浸润深度、肿瘤大小无相关性(P>0.05),而与组织分化程度相关,随着分化程度的增高,SLP-2的表达水平也有所增高(P=0.033)。3、宫颈鳞癌组中MTA1和SLP-2表达的相关性：宫颈鳞癌组中MTA1蛋白和SLP-2蛋白的表达呈负相关(r=-0.393,P=0.037); MTA1mRNA和SLP-2mRNA的表达水平亦成负相关(r=-0.327,P=0.025)。结论1、MTA1在宫颈鳞癌组织中高表达,其表达与肿瘤的淋巴结转移、肿瘤浸润深度密切相关,提示MTA1与宫颈鳞癌的发生发展有关,可作为判断宫颈鳞癌恶性程度的指标。2、SLP-2在宫颈鳞癌组织中低表达,与肿瘤分化程度相关,要明确SLP-2参与宫颈鳞癌发生发展的分子机制还有待进一步研究。3、MTA1和SLP-2在宫颈鳞癌中的表达呈负相关,MTA1高表达和SLP-2低表达可能与宫颈鳞癌发生发展有关。更多还原

【Abstract】 Background and ObjectiveCervical carcinoma is the most common malignant tumors in female bodies, and the incidence rate of which is slightly higher than breast cancer. Every year83%of new cases are in developing countries, with the tendency that the age is getting younger. Of all the cases, cervical squamous cell carcinoma is the most common type, this situation being severe, thus threatening the health and life of the female in our country. The continuous infection of High-risk Human papilla virus (HPV) makes up the major cause of cervical carcinoma; however, it is not the only reason. The incidence and development of it is the result of various reasons, with several phases and steps, on this account, indicators concerning clinical significance should be consulted. Recently researches have found that metastasis-associated gene1(MTA1) is a sort of gene related to tumor metastasis; also, researches on the SLP-2(stomatin-like protein2) protein to various tumor have found that to various tumors of proper pairing, the expression of SLP-2in tumor tissue is Overexpressed, while there exists not much research of it in cervical carcinoma tissue. As a result, this paper focuses on the following issues:whether the expression in tumor tissue is upexpressed or downexpressed; the relationship between SLP-2and clinic-pathological parameters, and whether SLP-2is relative to MTA1.Materials and methods: 1.Specimen origin:All samples are collected from the surgical excision tissue from May of2011to May of2012in the Second Affiliated Hospital of Zhengzhou University. The samples are divided as following:48cases of normal cervical tissue of total hysterectomy of uterine leiomyoma type(Group A);43cases of cervical intraepithelial neoplasia (CIN)(Group B);42cases of confirming pathological diagnosis of squamous carcinoma of the cervix without radiotherapy and chemotherapy before surgery(Group C). Samples of corresponding archival paraffin blocks in pathology department are also collected.2.Experimental methods:immunohistochemistry techniques is adapted to detect the expression and distribution of MTA1and SLP-2in different cervical tissues. The method of RT-PCR to detect the changes in MTAlmRNA and SLP-2mRNA levels.3.Statistical methods:SPSS17.0software package is adapted to analysis the data: both t test and one factor analysis of variance are adapted to examine the comparison of quantitative data,χ2to the comparison of rate, and2×2Correlation Analysis of paired data to relevance analysis. Inspection level:α=0.05(bilateral).Results:1.the expression of MTA1:in normal cervical tissues MTA1protein is mainly located in cell nucleuses while for CIN and MTA1in cervical carcinoma various protein location are found in the cell membranes, nucleuses and cytoplasm. The positive expression in Group A,B and C is overexpressed, and to same goes to the expression of MTAlmRNA. However the expression of latter in Group A is remarkably lower than Group B (P<0.05) and Group C (P<0.05). In Group B, the expression of MTA1is lower than Group C, but it has no statistical signification (P>0.05). the expression of MTA1in squamous carcinoma of the cervix has no relation to the age of the patient, clinical stages, degree of tissue differentiation, and the size of tumor (P>0.05), while it is highly related to lymphatic metastasis and the depth of myometrial invasion (P<0.05). the expression of MTA1will be upexpressed with the lymphatic metastasis of the patient and the deepening of the depth of myometrial invasion.2.The expression of SLP-2:in normal cervical tissues SLP-2is mainly located in cell membranes and cytoplasm, while for CIN and SLP-2protein in cervical carcinoma is mainly expressed in the form of cytoplasm staining. The positive expression of SLP-2protein is downexpressed in Group A, B and C, and the same goes to the expression of SLP-2mRNA. The expression of the latter in Group C is obviously lower than that of Group A(P<0.05) and B (P<0.05). In Group B, the expression of SLP-2is obviously lower than Group A(P<0.05). The expression of SLP-2bears no relations with the age of the patient, clinical stages, the existence of lymphatic metastasis, the depth of myometrial invasion, and the size of tumor (P>0.05), while it is relative to the degree of tissue differentiation. With the increase of the degree of differentiation, the espression level of SLP-2is also increase (P=0.033).3.The relativity of expression of MTA1and SLP-2in cervical carcinoma tissues: the expression of expression of MTA1and SLP-2in cervical carcinoma tissues is of inverse correlation(r=-0.393，P=0.037).Conclusion:1.The MTA1in cervical squamous cell carcinoma is of high expression, which is highly related to the lymphatic metastasis of tumors and the depth of myometrial invasion. The fact that MTA1is related to the occurrence and development of cervical carcinoma could be used as the index of the degree of malignancy of cervical squamous cell carcinoma.2.The SLP-2in cervical squamous cell carcinoma is of low expression, which is probably a new cancer suppressor gene. It is relative to the degree of tissue differentiation, however, further research is required to confirm the molecular mechanism of SLP-2in the occurrence and development of cervical squamous cell carcinoma.3.The expression of MTA1and SLP-2in cervical squamous cell carcinoma is of negative correlation, the high expression of MTAl and low expression of SLP-2is probably highly related to the occurrence and development of cervical squamous cell carcinoma as well as the biological behaviour.更多还原