T细胞浸润在肺腺癌发生中的意义以及与MicroRNA21的关系

发布时间：2018-08-23 12:57

【摘要】：背景肺癌的发生机制迄今尚未明了,是导致临床上对肺癌的防治不力的原因之一,因此肺癌成为危害人们身体健康的重大危险疾病。信号传导与转录激活因子3(Signal transducers and activators of transcription,STAT)是生物体内许多生长因子和细胞因子的主要信号传导蛋白,它在细胞的分化、功能、免疫平衡以及肿瘤发生中起着重要的作用。MicroRNA21(miR-21)是一种非编码的小分子RNA,参与机体内生长代谢、发育与疾病,并且与多种肿瘤的发生、发展、侵袭、转移有关。近年来关于miRNAs与STAT家族成员之间的信号转导之间的相互关系研究越来越多,尤其miR-21和STAT3信号通路的相互作用机制得到大多数人的认同。那么Mi R-21/STAT3信号转导通路在肺癌发生发展中的意义如何?以及对肿瘤内环境中,T淋巴细胞浸润的影响如何?对临床上治疗肺腺癌有何意义?目前尚未见公开报道。本课题通过采用实时荧光定量PCR(quantificational real-time polymerase chain reaction,q RT-PCR)等方法,检测人肺腺癌组织中STAT3、miR-21的表达情况,通过免疫组化检测肺腺癌组织中T4、T8细胞的浸润情况,分析二者之间的关系,探讨miR-21/STAT3信号转导通路异常与肺腺癌发生发展以及T细胞浸润之间的影响以及临床意义,为临床上进一步防治肺癌提供新思路。目的通过检测STAT3和miR-21在肺腺癌发生发展过程中的动态表达,肿瘤内环境中T4、T8细胞的浸润情况,探讨miR-21/STAT3在肺腺癌发生和对肿瘤内环境中淋巴细胞的浸润之间的作用与意义。方法所有标本均来自于2013年6月到2015年6月间,解放军第91中心医院检验病理科、河南焦作市人民医院病理科,手术切除的46例肺腺癌组织和癌旁组织,其中有34例转移癌;12例正常肺组织做为阴性对照。所取的肺腺癌组织、癌旁组织(需癌组织边缘以外约2-5cm)及正常的肺组织均为新鲜的组织标本,并将其放置于-80℃冰箱或液氮中进行保存。肺腺癌组织、癌旁组织及正常肺组织均经过高级职称病理医师确诊。实验所收集的上述标本不包括手术前已经过化学药物治疗的肺腺癌患者或复发的肺腺癌患者。采取q RT-PCR方法检测miR-21及STAT3基因表达情况,免疫组化方法检测T4、T8细胞的表达情况。采用SPSS17.0软件对实验结果进行t检验及单因素方差分析等相关统计分析。结果1常规病理诊断在此次收集的肺腺癌组织共46例,其中发生淋巴结转移癌34例;12例正常肺组织作为对照。所有组织均经过病理诊断。2 STAT3mRNA表达通过q RT-PCR方法检测STAT3mRNA在上述组织中的表达结果如下:正常肺组织为0.319±0.015;癌旁组织为0.631±0.020;肺腺癌组织为1.046±0.042;转移癌组织为1.235±0.015。正常肺组织、肺癌旁组织、肺腺癌组织、转移癌组织,组内之间基因表达的差异具有显著性(P0.01),肺腺癌组织中STAT3基因表达高于正常及癌旁组织。其中,肺腺癌与和正常肺组织比较,两者之间具有极显著性差异(P0.01)。3 miR-21的表达情况我们通过运用qRT-PCR方法检测了,miR-21在上述各类组织中的表达情况,实验结果显示如下:正常组0.0018±0.001;癌旁组0.017±0.005;腺癌组0.098±0.036;转移癌组0.126±0.134,组间方差分析F=17.016,P0.05,正常肺组织和肺腺癌组织之间存在极显著性差异(P0.01),肺腺癌与转移癌之间存在显著性差异(P0.05)。正常肺组织中miR-21呈现低表达,肺腺癌组织中miR-21呈现高表达。4 CD3+、CD4+、CD8+淋巴细胞在肿瘤组织中的占比情况肿瘤组织中CD3+细胞数量占比在正常组织中为76.12%,癌旁组与正常组比较,没有明显改变,腺癌组则出现明显降低;CD4+细胞数量占比在正常组中为45.17%,癌旁和腺癌组均低于正常组,具有显著性差异P0.05;CD8+细胞数量在正常组中为28.78%,腺癌组高于正常组,具有显著性差异P0.05。结论肺腺癌的发生与演进可能与miR-21升高以及体内免疫功能紊乱有关。
[Abstract]:BACKGROUND The pathogenesis of lung cancer is still unknown, which is one of the causes of poor clinical prevention and treatment of lung cancer. Lung cancer has become a major dangerous disease endangering people's health. Signal transducers and activators of transcription (STAT) are many growth factors and thin cells in organisms. MicroRNA21 (microRNA21) is a non-coding small molecule RNA involved in the body's growth, metabolism, development and disease, and is associated with the occurrence, development, invasion and metastasis of a variety of tumors. The relationship between signal transduction among members of STAT family is becoming more and more interrelated, especially the interaction mechanism between microRNAs-21 and STAT3 signaling pathways is widely recognized. What is the significance of the Mi R-21/STAT3 signaling pathway in the development of lung cancer and the effect on T lymphocyte infiltration in the tumor environment? In this study, the expression of STAT3 and microRNA-21 in human lung adenocarcinoma was detected by quantitative real-time polymerase chain reaction (q RT-PCR), and the immersion of T4 and T8 cells in lung adenocarcinoma was detected by immunohistochemistry. Objective To investigate the relationship between the expression of STAT3 and microRNAs-21 in lung adenocarcinoma and the development of lung adenocarcinoma and the infiltration of T cells. Methods From June 2013 to June 2015, 46 specimens were collected from the Department of Laboratory Pathology, 91 Central Hospital of PLA, Department of Pathology, Jiaozuo People's Hospital, Henan Province. Lung adenocarcinoma tissues and paracancerous tissues, including 34 cases of metastatic carcinoma and 12 cases of normal lung tissues as negative control. The lung adenocarcinoma tissues, paracancerous tissues (about 2-5 cm away from the margin of the tumor tissue) and normal lung tissues were fresh tissue specimens and stored in refrigerator or liquid nitrogen at - 80 C. Lung adenocarcinoma tissues, paracancerous tissues The specimens collected did not include patients with lung adenocarcinoma who had undergone chemotherapy before operation or patients with recurrent lung adenocarcinoma. Results 1 46 cases of lung adenocarcinoma tissues were collected by routine pathological diagnosis, including 34 cases of lymph node metastatic carcinoma, 12 cases of normal lung tissues as control. All tissues were pathologically diagnosed. 2 STAT3 mRNA expression was detected by Q RT-PCR. The results of AT3 mRNA expression in the above tissues were as follows: normal lung tissue was 0.319+0.015; paracancerous tissue was 0.631+0.020; lung adenocarcinoma tissue was 1.046+0.042; metastatic cancer tissue was 1.235+0.015. There were significant differences in gene expression between normal lung tissue, para-lung cancer tissue, lung adenocarcinoma tissue and metastatic cancer tissue (P 0.01). The expression of STAT3 gene in lung adenocarcinoma was significantly higher than that in normal and adjacent tissues (P 0.01). 3 The expression of microRNA-21 in lung adenocarcinoma and normal lung tissues was detected by qRT-PCR. The results showed that the expression of microRNA-21 in all kinds of tissues in normal group was 0.0018 (+0.001). There were significant differences between normal lung tissues and lung adenocarcinoma tissues (P 0.01). There were significant differences between lung adenocarcinoma and metastatic carcinoma tissues (P 0.05). The expression of microRNA-21 in normal lung tissues was low, and that in lung adenocarcinoma tissues was high (P 0.05). The proportion of CD3 +, CD4 +, CD8 + lymphocytes in tumor tissues was 76.12% in normal tissues. The number of CD3 + cells in tumor tissues was 76.12% in paracancerous group and normal group. There was no significant change in paracancerous group, but the number of CD4 + cells in adenocarcinoma group was significantly decreased. The proportion of CD4 + cells in normal group was 45.17%, and that in paracancerous and adenocarcinoma group was lower than normal group. The number of CD8 + cells in normal group was 28.78%, and that in adenocarcinoma group was higher than that in normal group, P 0.05. Conclusion The occurrence and evolution of lung adenocarcinoma may be related to the increase of microRNA-21 and the disturbance of immune function in vivo.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R734.2

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