【作者】 刘丽萍；

【导师】 雷冬梅；

【作者基本信息】 郑州大学 ， 妇产科学， 2021， 硕士

【摘要】 宫颈癌(Cervical cancer,CC)是全球女性最常见的妇科恶性肿瘤之一,其发病率和死亡率均排名第四,全球每年有近26.6万人死于该病,严重危害了人类健康和生命。在我国,宫颈癌的发病率主要集中在中西部地区,农村高于城市,并且呈现年轻化趋势,2001年FIGO流行病学调查报告显示,年轻妇女患宫颈癌的比例由9%上升到了24%。持续高危型人乳头瘤病毒(Human papillomavirus,HPV)感染是宫颈癌发生的主要因素,超过80%的女性会感染HPV,其中90%的感染患者体内病毒可被免疫系统所清除,10%发生持续感染,最终有1%进展为宫颈癌。目前有关宫颈肿瘤的具体发展机制尚不完全清楚,除了持续性HPV感染,有研究表明自噬失调及免疫因素异常也是导致恶性肿瘤发生发展的重要原因,酵母自噬相关基因6(Beclin 1)是参与自噬调控的重要基因之一,自噬在肿瘤的发展过程中起双重作用。程序性死亡受体配体-1(Programmed death ligand 1,PD-L1)是一种免疫共信号分子,可使肿瘤细胞发生免疫逃逸,二者均与肿瘤的发生发展密切相关,但有关二者在宫颈肿瘤中的具体作用机制尚不明确。本研究通过免疫组化SP法检测Beclin 1蛋白及PD-L1蛋白在正常宫颈组织、宫颈高级别鳞状上皮内病变(High grade squamous intraepithelial lesion,HSIL)及宫颈癌中的表达,结合临床数据分析在宫颈肿瘤发生发展过程中的作用及二者间的关系,为宫颈肿瘤发展机制的研究提供有价值的参考依据。目的通过免疫组化(Immunohistochemistry,IHC)SP法检测Beclin 1蛋白及PD-L1蛋白在宫颈组织中的表达,探究两者的相关性及临床意义。材料与方法1研究对象选取2018年10月至2019年12月在郑州大学第三附属医院因宫颈高级别鳞状上皮内病变行宫颈冷刀锥切术或因宫颈癌行广泛性子宫切除术的宫颈石蜡标本组织100例为实验组,同期因子宫肌瘤、子宫腺肌病、子宫脱垂等行子宫切除术的正常宫颈石蜡标本组织34例作为对照组。2实验方法应用免疫组化SP法分别检测各组宫颈石蜡标本组织中Beclin 1蛋白及PD-L1蛋白的表达。3统计学分析采用SPSS 22.0软件进行统计学分析,采用卡方检验或Fisher精确检验计算Beclin 1及PD-L1蛋白在不同组织中的表达及其与临床病理参数的关系,二者的相关性采用Spearman等级相关性分析,以P<0.05为差异有统计学意义。结果1三组宫颈组织中Beclin 1及PD-L1蛋白的表达情况Beclin1蛋白宫颈组织中的表达随病变的严重程度的增加而降低,即在正常宫颈组织中呈高表达,而在宫颈癌组织中低表达或不表达;PD-L1蛋白在宫颈组织中的表达随病变的严重程度的增加而升高,PD-L1在正常宫颈组织中不表达,在宫颈癌组织中呈高表达。2 Beclin 1及PD-L1蛋白的表达与宫颈癌临床病理参数的关系将宫颈癌组织中各种蛋白的表达情况与临床病理参数进行统计发现,Beclin1蛋白的表达与患者年龄、FIGO分期、组织浸润深度等无关,而与患者是否伴有HPV感染、肿瘤分化程度和淋巴结转移情况有关,差异具有统计学意义(P<0.05)。PD-L1蛋白的表达与患者年龄、肿瘤浸润深度及是否有淋巴结转移无关,而与患者是否伴有HPV感染、FIGO分期、肿瘤分化程度有关,差异有统计学意义(P<0.05)。3 Beclin 1与PD-L1蛋白表达的相关性分析通过Spearman相关性分析发现,Beclin 1与PD-L1蛋白表达呈负相关,差异具有统计学意义(r=-0.55,P<0.05)。结论Beclin1蛋白表达随宫颈病变严重程度的增加而降低,PD-L1蛋白表达随宫颈病变严重程度的增加而升高,二者呈负相关。更多还原

【Abstract】 Cervical cancer is one of the most common gynecological malignancies in women all over the world.The morbidity and mortality of the cervical cancer among all tumors is NO.4.It caused 266,000 deaths every year.In China,the incidence of cervical cancer is concentrated in the central and western regions mainly.The rate of the rural area is higher than the urban area,and the trend is younger.According to the2001 FIGO epidemiological survey report,it showed that the rate of cervical cancer of the young women that under 35 years old increased from 9% to 24%.As we all know,the type of the high-risk human papilloma virus(high-risk human papilloma virus,HPV)infection is the major reason which caused the happen of the cervical cancer.There are more than 80% of the women who may infect the HPV and 90% of the infected patients will be cleared by her immune system,10% persons may be infected persistently.Finally,there are only 1% of the patients will develope to the cervical cancer.At present,the specific development of the cervical cancer and the mechanisms involved is not fully clearly.In addition to the HPV infection,there are many studies have shown that the dysregulation of the autophagy and abnormal immune systems are also the important reasons for the occurrence and development of malignant tumors.Beclin 1 is one of the most important genes involved in the regulation of autophagy.It plays a dual role in tumor development.PD-L1 is an immune co-signaling molecule,which can cause the tumor immune escape of tumor cells.Both of them are closely to the occurrence and development of tumors,but the specific mechanisms of them in cervical tumors are not clear completely.From this research,we tested the expression of the Beclin 1 protein and the PD-L1 protein in the normal cervical tissue,the high level cervical squamous intraepithelial lesion(high level cervical squamous intraepithelial lesion,HSIL)and the cervical cancer respectively.Then we combined the results with the clinical data,and analyzed the role of both in cervical cancer and the relationship between them.From this experimence,we provide a valuable reference for the research on the developing mechanism of the cervical tumors.ObjectiveWe detected the expressions of Beclin 1 protein and PD-L1 protein in cervical tumor tissues through the immunohistochemical method,and then we analysed the correlation and clinical significance between them.Materials and methods1 Study subjectsWe choosed 100 cases of paraffin specimens of the cervical tumors as the experimental groups,which is come from the tissues of the cold knife cut cone because of the HSIL or do the hysterectomyfull because of the cervical cancer in The Third Affiliated Hospital of Zhengzhou University in October 2018 to December2019.At the same time,we choosed another 34 cases as the control group,which are come from the patient who did the operation of total hysterectomy because of the hysteromyoma,adenomyosis or the metroptosis.2 MethodsWe used the Immunohistochemical SP method(Immunohistochemistry,IHC)to test the expression of Beclin 1 and PD-L1 proteins in the paraffin tissue of each group.3 Statistical analysisWe used the SPSS 22.0 software to analyse the statistics.Chi-square test or Fisher’s exact test was used to calculate the expression of Beclin 1 and PD-L1 proteins and then we explored their relationship with clinicopathological parameters in different tissues.The p<0.05 indicates that the difference have statistically significant.Result1 Beclin1 and PD-L1 protein expression in different cervical lesions.The expression of Beclin1 protein is decreased along with the increase of the severity of pathological tissues,it is highly express in the normal cervical tissues,but is low or not express in cervical cancer tissues.The PD-L1 protein is not express in the normal cervical tissues,but it is highly express in the cervical cancer tissues,and it is increased along with the increase of the severity of pathological tissues.2 The relationship between the expression of the Beclin 1 and PD-L1 proteins and their clinicopathological parameters in the cervical cancer.The expression of the Beclin1 protein is not correlated with the age,clinicopathological stages,and the degree of the tissue invasion,but it is correlated with the HPV infection,the degrees of tumor differentiation,and the lymph node metastasis.The difference was statistically significant(p<0.05).And the expression of PD-L1 protein is not related to the age of patients,the depth of tumor invasion and the presence of lymph node metastasis.But it is related to the HPV infection,clinical stages and the degrees of tumor differentiation.The difference was statistically significant(p<0.05).3 The correlation analysis of Beclin 1 and PD-L1 proteins.The Spearman correlation analysis showed that Beclin 1 was negatively correlated with PD-L1 protein expression,and the difference was statistically significant(r=-0.55,p<0.05).ConclusionThe expression of Beclin1 protein decreased with the increase of the severity of cervical lesions,while the expression of PD-L1 protein increased with the increase of the severity of cervical lesions.Both of which were negatively correlated.更多还原