黑色素瘤相关抗原-A3、-A9在宫颈病变的表达及临床意义

发布时间：2018-06-22 10:26

本文选题：宫颈上皮内瘤变 + 宫颈癌　；参考：《河北大学》2017年硕士论文

【摘要】：目的:通过对病理诊断异常的宫颈病变组织检测MAGE-A3、MAGE-A9蛋白及基因的表达情况,并与正常组织进行对比。研究MAGE-A3、MAGE-A9与宫颈上皮内瘤变及宫颈癌的发生和发展的关系,从而对宫颈病变的诊断和治疗提供一定的理论依据,进而研究出切实可行的治疗方案。方法:选取我院2009年-2015年收治的宫颈病变患者,其中宫颈上皮内瘤变(CINⅢ)40例,宫颈癌Ia~b期(FIGO分期)60例,宫颈癌IIa期(FIGO分期)24例,正常宫颈40例。所有患者均进行手术治疗,查阅患者病历资料,结合患者的临床病理资料进行分析。调取所有的患者病理组织蜡块,行免疫组织化学染色,光镜下观察MAGE-A3、MAGE-A9表达情况。同时收集2015年-2016年我院手术室切下新鲜手术组织,运用实时荧光PCR方法检测:15例正常宫颈,15例CINⅢ,20例宫颈癌组织中,MAGE-A3、MAGE-A9基因的表达情况。运用Western Blot方法检测15例正常宫颈,15例CINⅢ组织,20例宫颈癌组织中MAGE-A3、MAGE-A9蛋白的表达情况。结果:1.免疫组化法显示:MAGE-A3蛋白主要定位在细胞质内,且表达水平随着宫颈病变程度的加重而逐渐升高。MAGE-A3在正常宫颈、CINⅢ和宫颈癌组织中的阳性表达率分别为5.00%、15.00%、48.81%,三者阳性表达呈逐渐上升的趋势,统计学分析发现,宫颈癌组分别与CINⅢ组及正常对照组相比,MAGE-A3的表达明显升高,均有显著性差异(P0.01);但是,正常组与CINⅢ组相比较,差异无统计学意义(P0.05);同时研究显示MAGE-A3的表达水平与宫颈癌的临床分期、病理分级、淋巴结转移及高危型HPV感染之间差异均有统计学意义(P0.05)。MAGE-A9蛋白主要定位在细胞质内,MAGE-A9在正常宫颈、CINⅢ和宫颈癌组织中的阳性表达率分别为7.50%、17.50%、55.95%,三者阳性表达呈逐渐上升的趋势,统计学分析发现,宫颈癌组分别与CINⅢ组及正常对照组相比,MAGE-A9的表达明显升高,均有显著性差异(p0.01);但是,正常组与CINⅢ组相比较,差异无统计学意义(P0.05);同时研究显示MAGE-A9的表达水平与宫颈癌的临床分期、各病理分级以及有无淋巴结转移之间差异均有统计学意义(p0.05)。2.实时荧光定量PCR法显示:(1)以GAPDH mRNA为内参照标准,正常宫颈、CINⅢ及宫颈癌组织中MAGE-A3 mRNA的相对表达水平分别为0.184±0.069,0.221±0.086与0.702±0.145。MAGE-A3mRNA的相对表达量在宫颈癌中明显高于CINⅢ及正常组织,宫颈癌组与正常组及CINⅢ组间差异均有统计学意义(P0.05),而正常组与CINⅢ组之间差异无统计学意义(P0.05)(2)以GAPDH mRNA为内参照标准,正常宫颈、CINⅢ及宫颈癌组织中MAGE-A9 mRNA的相对表达水平分别为0.182±0.062,0.234±0.087与0.642±0.150,其中宫颈癌组织中的MAGE-A9表达量明显高于CIN及正常组织,宫颈癌组与正常组及CINⅢ组间差异均有统计学意义(P0.05),而正常组与CINⅢ组之间差异无统计学意义(P0.05)3.Western bloting蛋白印迹显示:(1)MAGE-A3的灰度随着宫颈组织病变程度的加重逐渐加深,正常宫颈、CINⅢ及宫颈癌组织中MAGE-A3蛋白的相对表达水平分别为0.943±0.178、1.035±0.139、1.977±0.105。宫颈癌组织中的MAGE-A9蛋白的相对表达量明显高于CINⅢ及正常宫颈组织,差异均有统计学意义(P0.05),而正常组与CINⅢ组之间差异无统计学意义(P0.05)。(2)MAGE-A9的灰度随着宫颈组织病变程度的加重逐渐加深,正常宫颈、CIN及宫颈癌组织中MAGE-A9蛋白的相对表达水平分别为0.857±0.315、0.909±0.526、1.882±0.179。宫颈癌组织中的MAGE-A9蛋白的相对表达量明显高于CINⅢ及正常宫颈组织,差异均有统计学意义(P0.05),而正常组与CINⅢ组之间差异无统计学意义(P0.05)。结论:1、MAGE-A3的过表达与宫颈癌的发生发展紧密相关,而且MAGE-A3的过量表达与宫颈癌的临床分期,病理分级及淋巴转移及高危HPV感染紧密相关,而与年龄及癌症类型没有明显关联,MAGE-A3有可能成为宫颈疾病早期诊断、宫颈癌治疗的重要肿瘤标记物。MAGE-A3可能成为宫颈癌生物治疗提供理想的靶点,对于进一步了解宫颈癌生物学行为判断预后有很重要的临床应用价值。2、MAGE-A9的过表达与宫颈癌发生发展密切相关,且MAGE-A9的过量表达与宫颈癌的临床分期、病理分级及有无淋巴转移有关,MAGE-A9有可能成为宫颈疾病早期诊断、宫颈癌治疗的重要肿瘤标记物。MAGE-A9可能成为宫颈癌生物治疗提供理想的靶点,MAGE-A9对于进一步了解宫颈癌生物学行为判断预后有很重要的临床应用价值。
[Abstract]:Objective: to detect the expression of MAGE-A3, MAGE-A9 protein and gene in pathological diagnosis of cervical lesions, and to compare with normal tissues and study the relationship between MAGE-A3, MAGE-A9 and cervical intraepithelial neoplasia and the occurrence and development of cervical cancer, so as to provide a theoretical basis for the diagnosis and treatment of cervical lesions. Methods: a practical treatment scheme was taken. Methods: selected patients with cervical lesions admitted in our hospital in -2015 2009, 40 cases of cervical intraepithelial neoplasia (CIN III), 60 cases of Ia~b stage (FIGO staging) of cervical cancer, 24 cases of IIa stage (FIGO stage) of cervical cancer and 40 cases of normal cervix. All patients were treated with surgical treatment, consult the patient's medical records, combined with patient's presence. The pathological data of the bed were analyzed. All the pathological tissue wax blocks were collected, the immunohistochemical staining was performed and the expression of MAGE-A3 and MAGE-A9 was observed under light microscope. At the same time, the fresh surgical tissues were cut down in the operation room of our hospital in 2015 -2016, and the real time fluorescence PCR method was used to detect 15 cases of normal cervix, 15 cases of CIN III, and 20 cases of cervical cancer tissues, MAGE- The expression of A3, MAGE-A9 gene. The expression of MAGE-A3 and MAGE-A9 in 15 cases of normal cervix, 15 cases of CIN III tissue and 20 cases of cervical cancer were detected by Western Blot. Results: 1. immunohistochemistry showed that MAGE-A3 protein was mainly located in the cytoplasm, and the level of expression gradually increased with the severity of cervical lesions. The positive expression rate of A3 in normal cervix, CIN III and cervical cancer tissues was 5%, 15%, 48.81%, and the positive expression of three was gradually rising. The statistical analysis showed that the expression of MAGE-A3 in the cervical cancer group was significantly higher than that of the CIN group and the normal control group (P0.01), but the normal group and the CIN III group were significantly different. The difference was not statistically significant (P0.05). At the same time, the study showed that the expression level of MAGE-A3 and the clinical stage of cervical cancer, pathological classification, lymph node metastasis and high risk HPV infection were statistically significant (P0.05).MAGE-A9 protein was mainly located in the cytoplasm, MAGE-A9 was in the normal cervix, CIN III and cervical cancer tissues. The positive expression rates were 7.50%, 17.50%, 55.95% and three, respectively. The statistical analysis showed that the expression of MAGE-A9 was significantly higher in the cervical cancer group than in the CIN III group and the normal control group (P0.01), but there was no significant difference between the normal group and the CIN group (P0.05). The results showed that the expression level of MAGE-A9 and the clinical stage of cervical cancer, the pathological grades and the lymph node metastasis were statistically significant (P0.05).2. real-time fluorescence quantitative PCR method showed: (1) the relative expression level of MAGE-A3 mRNA in normal cervix, CIN III and cervical cancer tissues was 0.184 +, respectively, with GAPDH mRNA as the internal reference standard. The relative expression of 0.069,0.221 + 0.086 and 0.702 + 0.145.MAGE-A3mRNA in cervical cancer was significantly higher than that of CIN III and normal tissue. The difference between the cervical cancer group and the normal group and the group CIN III was statistically significant (P0.05), while the difference between the normal group and the CIN III group was not statistically significant (P0.05) (2) with GAPDH mRNA as the internal reference standard, normal cervix, CIN, and CIN. The relative expression level of MAGE-A9 mRNA in cervical cancer tissues was 0.182 + 0.062,0.234 + 0.087 and 0.642 + 0.150 respectively. The expression of MAGE-A9 in cervical cancer tissues was significantly higher than that of CIN and normal tissues. The difference between the cervical cancer group and the normal group and the CIN III group was statistically significant (P0.05), but there was no difference between the normal group and the CIN group group III group. The study significance (P0.05) 3.Western bloting Western blot showed: (1) the gray level of MAGE-A3 gradually deepened with the severity of cervical tissue lesion, and the relative expression level of MAGE-A3 protein in normal cervix, CIN III and cervical cancer tissues was relative expression of MAGE-A9 protein in 0.943 + 0.178,1.035 + 0.139,1.977 + 0.105. cervical cancer tissues The difference was significantly higher than that of CIN III and normal cervical tissue (P0.05), but there was no significant difference between the normal group and the CIN group (P0.05). (2) the gray level of MAGE-A9 gradually deepened with the severity of cervical tissue lesion, and the relative expression level of MAGE-A9 protein in normal cervix, CIN and cervical cancer tissues was 0.857 + 0, respectively. The relative expression of MAGE-A9 protein in.315,0.909 + 0.526,1.882 + 0.179. cervical cancer tissues was significantly higher than that of CIN III and normal cervical tissue (P0.05), but there was no significant difference between the normal group and the CIN group (P0.05). Conclusion: 1, the overexpression of MAGE-A3 is closely related to the development of cervical cancer, and the MAGE- is closely related to the development of cervical cancer, and MAGE- is closely related to the occurrence and development of cervical cancer, and MAGE-, and MAGE- The overexpression of A3 is closely related to the clinical stage of cervical cancer, pathological grading, lymphatic metastasis and high risk HPV infection, but not associated with age and cancer type. MAGE-A3 may be an early diagnosis of cervical disease. The important tumor marker for cervical cancer treatment can be an ideal target for the biological treatment of cervical cancer. To further understand the prognosis of cervical cancer biological behavior is of great clinical value.2, the overexpression of MAGE-A9 is closely related to the development of cervical cancer, and the overexpression of MAGE-A9 is related to the clinical stage of cervical cancer, pathological classification and lymphatic metastasis. MAGE-A9 may be the early diagnosis of cervical disease and cervical cancer. The important tumor marker,.MAGE-A9, may be an ideal target for the biological treatment of cervical cancer. MAGE-A9 is of great clinical value for the further understanding of the prognosis of cervical cancer biological behavior.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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