肿瘤相关成纤维细胞影响子宫内膜癌发展的研究

发布时间：2018-06-29 08:11

本文选题：肿瘤微环境 + 子宫内膜癌　；参考：《天津医科大学》2014年博士论文

【摘要】：子宫内膜癌是西方发达国家最常见的女性生殖道肿瘤,近年来随着饮食结构的西化,生育观念的变化,激素的使用,我国EC的发生率明显上升,且趋于年轻化。长期以来EC发病机制研究多以肿瘤细胞本身为靶点,认为肿瘤生成是大量肿瘤细胞自主生长,不受机体调控的过程,这一理论强调了肿瘤细胞本身的作用,而忽略了微环境与肿瘤细胞之间的相互作用。肿瘤微环境是一个由肿瘤细胞、基质细胞、细胞因子、细胞外基质等共同构成的肿瘤发生、发展和转移的局部病理环境。间质细胞包括癌相关成纤维细胞(cancer-associated fibroblasts,CAFs)、肿瘤相关巨噬细胞,内皮细胞、免疫细胞和炎症细胞等,其中最重要的是CAFs,其数量多,分布广,过去认为其仅扮演着支持、营养上皮细胞的"被动"角色,而近来研究发现,CAFs在调节肿瘤微环境的动态平衡中扮演着"主动"角色,甚至决定了肿瘤上皮的命运。CAFs可产生丰富的细胞因子,其中SDF-1及其受体CXCR4在肿瘤微环境中的作用逐渐引起人们重视。目前CAFs促进肿瘤生长的研究多集中在乳腺癌、肺癌、前列腺癌,而子宫内膜癌方面的研究甚少。故本研究拟从肿瘤微环境的角度,探讨其在子宫内膜癌中的发生发展及其机制。目的:体内外实验探讨子宫内膜癌CAFs对子宫内膜癌细胞生物学行为的影响,检测子宫内膜癌CAFs分泌SDF-1的情况,探讨SDF-1能否促进子宫内膜癌细胞生长、迁移和侵袭。方法:体外实验采用MTT法和Transwell小室法检测子宫内膜癌CAFs/正常子宫内膜NFs对子宫内膜癌ECC-1和HEC-1B细胞增殖、迁移和侵袭的影响,体内实验按照子宫内膜癌CAFs/正常子宫内膜NFs与子宫内膜癌细胞为1:3的混合比例(细胞总数为8×106个)接种裸鼠皮下移植瘤,观察成瘤时间、成瘤率、淋巴结转移率、肿瘤质量和肿瘤生长体积;体外实验采用Elisa法检测子宫内膜癌CAFs和正常子宫内膜NFs培养上清液中SDF-lα的水平;体外实验采用MTT法和Transwell小室法检测SDF-1α及CXCR4的拮抗剂AMD3100对子宫内膜癌ECC-1和HEC-1B细胞增殖、迁移和侵袭的影响;采用明胶酶谱法检测SDF-1α及AMD3100对子宫内膜癌ECC-1和HEC-1B细胞表达MMPs的影响。结果:(1)子宫内膜癌CAFs与子宫内膜癌混合组较正常子宫内膜NFs与子宫内膜癌混合组比较在混合比例为1:3时的72小时达促增殖高峰(P0.05),且子宫内膜癌CAFs与子宫内膜癌混合组较正常子宫内膜NFs与子宫内膜癌混合组比较促增殖作用强(P0.05)。(2)子宫内膜癌CAFs较正常子宫内膜NFs促进子宫内膜癌细胞的迁移和侵袭(P0.05)。(3)子宫内膜癌CAFs组、NFs组和空白组裸鼠移植瘤均未见淋巴结转移,成瘤率均为100%。(4)CAFs组裸鼠成瘤时间、肿瘤质量和肿瘤生长速率较NFs组及空白对照组均明显缩短(P0.05)。(5)子宫内膜癌CAFs细胞较正常子宫内膜NFs细胞分泌SDF-1α明显增多(P0.05)。(6)SDF-1α浓度大于200ng/ml时可增强子宫内膜癌细胞的增殖、迁移和侵袭能力(P0.05)。AMD3100浓度大于500ng/ml时可阻断SDF-1α对子宫内膜癌细胞的增殖、迁移和侵袭作用(P0.05)。(7)200ng/ml的SDF-1α可促进子宫内膜癌细胞表达 MMP-2 和 MMP-9(P0.05),500ng/ml 的 AMD3100 可阻断 SDF-1α 对子宫内膜癌细胞高表达MMP-2和MMP-9的作用(P0.05)。结论:子宫内膜癌CAFs分泌SDF-1α增多,SDF-1α可通过SDF-1/CXCR4生物轴促进子内膜癌细胞的增殖、迁移、侵袭及MMP-2和MMP-9的表达。
[Abstract]:Endometrial carcinoma is the most common female reproductive tract tumor in western developed countries. In recent years, with the Westernization of the diet structure, the change of the concept of fertility and the use of hormone, the incidence of EC in our country has risen obviously and tends to be young. For a long time, the pathogenesis of EC is mostly targeted by the tumor cell itself, and it is considered that tumor formation is a large number of tumors. This theory emphasizes the role of the tumor cell itself and neglects the interaction between the microenvironment and the tumor cells. The tumor microenvironment is a local pathological environment, which consists of tumor cells, matrix cells, cytokines, and extracellular matrix. Interstitial cells include cancer related fibroblasts (cancer-associated fibroblasts, CAFs), tumor related macrophages, endothelial cells, immune cells and inflammatory cells. The most important of which is CAFs, which has a large number and wide distribution. In the past, it was thought to play the role of the "passive" role of nutritional epithelial cells, and recent studies have found that CAFs is in the form of CAFs. The dynamic balance of the tumor microenvironment plays a "active" role, and even determines the fate of the tumor epithelium.CAFs can produce abundant cytokines, in which the role of SDF-1 and its receptor CXCR4 in the tumor microenvironment is gradually aroused people's attention. At present, the research of CAFs for promoting tumor growth is mainly concentrated in breast, lung, and prostate cancer. The study of endometrial carcinoma is very rare. Therefore, this study intends to explore its development and mechanism in endometrial carcinoma from the perspective of tumor microenvironment. Objective: To investigate the effect of endometrial carcinoma CAFs on the biological behavior of endometrial carcinoma cells in vivo and in vitro and to detect the secretion of SDF-1 in endometrial carcinoma CAFs, and to explore the ability of SDF-1. Methods: MTT method and Transwell chamber method were used to detect the effect of NFs on the proliferation, migration and invasion of endometrial carcinoma ECC-1 and HEC-1B cells in endometrial carcinoma with MTT and Transwell compartment method in vitro. In vivo experiment, endometrial carcinoma CAFs/ normal endometrium NFs and endometrial carcinoma The cells were inoculated subcutaneously in nude mice with a mixed proportion of 1:3 (8 x 106 cells). The tumor formation time, tumor formation rate, lymph node metastasis rate, tumor mass and tumor growth volume were observed. The level of SDF-l alpha in endometrial carcinoma CAFs and normal endometrium NFs cultured supernatant was detected by Elisa method in vitro; in vitro experiment was carried out by MTT method. The effects of SDF-1 alpha and CXCR4 antagonist AMD3100 on proliferation, migration and invasion of endometrial carcinoma ECC-1 and HEC-1B cells were detected with Transwell cell method. The effect of SDF-1 A and AMD3100 on ECC-1 and HEC-1B cell expression MMPs in endometrial carcinoma was detected by gelatinase method. Results: (1) endometrial carcinoma CAFs and endometrial carcinoma were more normal. Endometrial NFs was compared with endometrial carcinoma in a mixed group of 72 hours when the mixing ratio was 1:3 (P0.05), and endometrial carcinoma CAFs and endometrial carcinoma group had stronger proliferation promoting effect than normal endometrium NFs and endometrial carcinoma group (P0.05). (2) endometrial carcinoma CAFs was better than normal endometrium NFs to promote uterus. The migration and invasion of endometrial carcinoma cells (P0.05). (3) CAFs group of endometrial carcinoma, no lymph node metastasis was found in group NFs and nude mice, and the tumor formation rate was 100%. (4) CAFs group, and the tumor quality and growth rate were significantly shorter than that of NFs group and blank control group (P0.05). (5) endometrial carcinoma CAFs cells were more than normal children. The secretion of SDF-1 a in endometrium NFs cells (P0.05). (6) the proliferation of endometrial carcinoma cells can be enhanced when the concentration of SDF-1 alpha is greater than 200ng/ml, and the migration and invasion ability (P0.05).AMD3100 concentration is greater than 500ng/ml, which can block the proliferation, migration and invasion of SDF-1 alpha to endometrial cancer cells (P0.05). (7) 200ng/ml SDF-1 alpha can promote the uterus. Endometrial carcinoma cells express MMP-2 and MMP-9 (P0.05), AMD3100 of 500ng/ml can block the high expression of MMP-2 and MMP-9 in endometrial carcinoma cells (P0.05). Conclusion: CAFs secreted SDF-1 alpha in endometrial carcinoma. SDF-1 alpha promotes the proliferation, migration, invasion, and tables of endometrial carcinoma cells through the biological axis. Da.
【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33

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