RIPK3介导的巨噬细胞M2型极化在结直肠癌肝转移中的研究

高辉; 殷玉敬; 孙志慧

doi:10.16190/j.cnki.45-1211/r.2023.01.012

RIPK3介导的巨噬细胞M2型极化在结直肠癌肝转移中的研究

RIPK3-mediated M2-type polarization of macrophages in colorectal cancer liver metastasis

摘要

摘要: 目的：探究受体相互作用丝氨酸/苏氨酸激酶3（RIPK3）介导的巨噬细胞M2型极化在结直肠癌肝转移（CRLM）中的作用及其相关机制。方法：收集CRLM患者结肠肿瘤及其癌旁组织，对肿瘤组织进行病理分析。通过RT-qPCR法检测RIPK3、CD68、精氨酸酶-1（Arg-1）表达量。采集CRLM患者和结肠癌患者外周血，通过流式细胞术检测巨噬细胞分型，利用ELISA检测血清白介素（IL）-1β、IL-6、诱导型一氧化氮合酶（iNOS）、IL-4、转化生长因子-β（TGF-β）、IL-10 含量。使用慢病毒将PLKRIPK3和PLK-NC转染至THP-1细胞，采用PMA/IL4诱导为M2型巨噬细胞，通过RT-qPCR和Western blotting检测两组细胞极化程度。将两组细胞分别与人结肠癌细胞系WiDr共孵育，检测WiDr细胞迁移、侵袭变化和结肠癌相关转移1（MACC1）、原癌基因MYC（c-MYC）表达情况。结果：CRLM患者结肠肿瘤病理结构符合肠腺癌肝转移。与癌旁组织相比，RIPK3在CRLM肿瘤组织表达降低，CD68、Arg-1表达增加。与结肠癌患者相比，CRLM患者外周血中富集更多的M2型巨噬细胞，血清中IL-4、TGF-β、IL-10含量增加，IL-1β、iNOS含量减少。与WiDr+PLK-NC-THP-1组相比，WiDr+PLK-RIPK3-THP-1组被诱导为M2型巨噬细胞的程度减少，对WiDr细胞的促迁移和侵袭能力降低，MACC1、c-MYC表达量减少。结论：RIPK3表达降低通过诱导巨噬细胞向M2型极化，促进结直肠癌细胞MACC1、c-MYC表达和肿瘤转移。

Abstract: Objective: To study the role of M2-type polarization in macrophages mediated by receptor interacting serine/threonine kinase 3 (RIPK3) in colorectal cancer liver metastasis (CRLM) and its related mechanisms.Methods: Colon tumors and their paracancer tissues were collected from CRLM patients for pathological analysis.The expressions of RIPK3, CD68 and Arg-1 were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR).The peripheral blood of CRLM and colon cancer patients was collected, and the macrophage typing was detected by flow cytometry.Serum levels of interleukin-1β, IL-6, inducible nitric oxide synthase (iNOS), IL-4, transforming growth factor-β (TGF-β) and IL-10 were detected by ELISA.PLK-RIPK3 and PLK-NC were transfected into THP-1 cells by lentivirus, and M2-type macrophages were induced by PMA/IL4.The polarization degree of the two groups of cells was detected by RT-qPCR and western blotting.The two groups of cells were co-incubated with human colon cancer cell line WiDr, respectively; the migration, invasion changes and expression of metastasis associated in colon cancer 1 (MACC1) and MYC proto-oncogene (c-MYC) of WiDr cells were detected.Results: The pathological structure of colon tumors in CRLM patients was consistent with liver metastasis of intestinal adenocarcinoma.Compared with paracancer tissues, the expression of RIPK3 in CRLM tumor tissues decreased while the expressions of CD68 and Arg-1 increased.Compared with colon cancer patients, the peripheral blood of CRLM patients was more enriched with M2-type macrophages; the serum contents of IL-4, TGF-β and IL-10 increased while the contents of IL-1β and iNOS decreased.Compared with the WiDr+PLK-NC-THP-1 group, the WiDr+PLK-RIPK3-THP-1 group was less induced into M2 macrophages; the ability to promote migration and invasion of WiDr cells and the expressions of MACC1 and c-MYC decreased.Conclusion: The expression of reduced RIPK3 promotes the expressions of MACC1 and c-MYC and tumor metastasis in colorectal cancer cells by inducing M2-type polarization of macrophages.

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