双调蛋白过表达促进小鼠肠癌CT26细胞的体内生长

发布时间：2018-05-21 13:31

本文选题：肿瘤微环境 + 双调蛋白　；参考：《肿瘤》2017年07期

【摘要】：目的:探讨上皮细胞生长因子双调蛋白(amphiregulin,AREG)对小鼠肠癌CT26细胞生长的影响及其相关机制。方法:采用ELISA法检测不同小鼠肿瘤细胞中AREG蛋白的表达水平。将携带AREG基因的重组慢病毒载体转染至小鼠肠癌CT26细胞、黑素瘤B16细胞和肝癌LPC-Akt细胞,同时以转染空载体细胞作为阴性对照。采用MTT法、克隆形成实验和FCM法分别检测AREG过表达后细胞的体外增殖和克隆形成能力以及细胞周期进程。构建AREG过表达CT26细胞的小鼠移植瘤模型,观察小鼠体内移植瘤的生长情况,并且采用FCM法和实时荧光定量PCR法分别检测移植瘤组织中免疫细胞的分布情况以及趋化因子的表达水平。结果:小鼠肠癌CT26、黑素瘤B16和肝癌LPC-Akt细胞中AREG相对低表达。过表达AREG后,上述3种肿瘤细胞的体外增殖能力、克隆形成能力及细胞周期进程均无明显变化(P值均0.05)。然而,在CT26细胞的小鼠移植瘤模型中,AREG过表达可明显促进肿瘤细胞的体内生长(P0.01),下调肿瘤组织中CD8~+T细胞所占比例(P0.05),并降低与CD8+T细胞招募相关的CC趋化因子配体5(CC chemokine ligand 5,CCL5)的转录水平(P0.05)。结论:AREG能够促进小鼠肠癌CT26细胞的体内生长,推测其作用机制可能与调控CD8+T细胞招募相关趋化因子,从而影响肿瘤微环境有关。
[Abstract]:Aim: to investigate the effects of epithelial growth factor (EGF) bimodontin on the growth of CT26 cells in mouse colorectal cancer and its related mechanisms. Methods: ELISA assay was used to detect the expression of AREG protein in different mouse tumor cells. The recombinant lentivirus vector carrying AREG gene was transfected into mouse colorectal cancer CT26 cells, melanoma B16 cells and hepatocellular carcinoma LPC-Akt cells. MTT assay, clone formation assay and FCM assay were used to detect the proliferation and clone formation ability and cell cycle process of AREG overexpression cells in vitro, respectively. A mouse model of transplanted tumor with overexpression of AREG CT26 cells was established to observe the growth of transplanted tumor in mice. The distribution of immunocytes and the expression of chemokines were detected by FCM and real-time quantitative PCR. Results: the expression of AREG in mouse colorectal cancer CT26, melanoma B 16 and hepatoma LPC-Akt cells was relatively low. After overexpression of AREG, the proliferative ability, clone formation ability and cell cycle progression of the above three tumor cells showed no significant change (P = 0.05). However, In mouse transplanted tumor model of CT26 cells, the overexpression of Arg can significantly promote the growth of tumor cells in vivo, down-regulate the proportion of CD8 ~ T cells in tumor tissues and decrease the CC chemokine ligand 5(CC chemokine associated with CD8 T cell recruitment. The transcription level of ligand 5 (CCL5) is P0.05. Conclusion the growth of mouse colorectal cancer CT26 cells can be promoted by WAREG, and the mechanism may be related to the regulation of recruitment of chemokines by CD8 T cells, thus affecting the tumor microenvironment.
【作者单位】：上海交通大学医学院附属仁济医院上海市肿瘤研究所癌基因及相关基因国家重点实验室;
【基金】：国家自然科学基金资助项目(编号:81402348)~~
【分类号】：R73-3

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