本文選題：腫瘤微環(huán)境 + 雙調(diào)蛋白�。� 參考：《腫瘤》2017年07期

【摘要】：目的:探討上皮細(xì)胞生長(zhǎng)因子雙調(diào)蛋白(amphiregulin,AREG)對(duì)小鼠腸癌CT26細(xì)胞生長(zhǎng)的影響及其相關(guān)機(jī)制。方法:采用ELISA法檢測(cè)不同小鼠腫瘤細(xì)胞中AREG蛋白的表達(dá)水平。將攜帶AREG基因的重組慢病毒載體轉(zhuǎn)染至小鼠腸癌CT26細(xì)胞、黑素瘤B16細(xì)胞和肝癌LPC-Akt細(xì)胞,同時(shí)以轉(zhuǎn)染空載體細(xì)胞作為陰性對(duì)照。采用MTT法、克隆形成實(shí)驗(yàn)和FCM法分別檢測(cè)AREG過(guò)表達(dá)后細(xì)胞的體外增殖和克隆形成能力以及細(xì)胞周期進(jìn)程。構(gòu)建AREG過(guò)表達(dá)CT26細(xì)胞的小鼠移植瘤模型,觀察小鼠體內(nèi)移植瘤的生長(zhǎng)情況,并且采用FCM法和實(shí)時(shí)熒光定量PCR法分別檢測(cè)移植瘤組織中免疫細(xì)胞的分布情況以及趨化因子的表達(dá)水平。結(jié)果:小鼠腸癌CT26、黑素瘤B16和肝癌LPC-Akt細(xì)胞中AREG相對(duì)低表達(dá)。過(guò)表達(dá)AREG后,上述3種腫瘤細(xì)胞的體外增殖能力、克隆形成能力及細(xì)胞周期進(jìn)程均無(wú)明顯變化(P值均0.05)。然而,在CT26細(xì)胞的小鼠移植瘤模型中,AREG過(guò)表達(dá)可明顯促進(jìn)腫瘤細(xì)胞的體內(nèi)生長(zhǎng)(P0.01),下調(diào)腫瘤組織中CD8~+T細(xì)胞所占比例(P0.05),并降低與CD8+T細(xì)胞招募相關(guān)的CC趨化因子配體5(CC chemokine ligand 5,CCL5)的轉(zhuǎn)錄水平(P0.05)。結(jié)論:AREG能夠促進(jìn)小鼠腸癌CT26細(xì)胞的體內(nèi)生長(zhǎng),推測(cè)其作用機(jī)制可能與調(diào)控CD8+T細(xì)胞招募相關(guān)趨化因子,從而影響腫瘤微環(huán)境有關(guān)。
[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.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院上海市腫瘤研究所癌基因及相關(guān)基因國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(編號(hào):81402348)~~
【分類(lèi)號(hào)】：R73-3

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