Objective: To investigate the effects of different expression of monoacylglycerol lipase (MAGL) in tumor-associated macrophages (TAMs) with the proliferation of MHCC97H human liver cancer cells in vivo and its mechanism. Methods: Human peripheral blood-derived monocyte was induced to differentiate into M2-type TAMs and was identified by flow cytometry. The co-culture model of TAMs and MHCC97H human liver cancer cells was established, and the expression of MAGL in TAMs cells was detected by qRT-PCR. The expression of MAGL in TAMs cells was detected by plasmid transfection. ELISA and qRT-PCR was used to detect the mRNA expression levels and secretion levels of inflammatory factors in TAMs cells. The subcutaneous tumor model of MHCC97H mice was constructed to observe the effect of different expression of MAGL in TAMs cells with the proliferation of MHCC97H human liver cancer cells in vivo. F-test was used for the measurement of homogeneity of variance between two independent samples. A t-test was used for homogeneity of variance, and the corrected t-test was used for non-homogeneity of variance. Results: Human peripheral blood-derived monocytes were successfully induced to differentiate into M2-type TAMs. An in vitro co-culture model was established. qRT-PCR showed that MHCC97H human liver cancer cells significantly down-regulated the expressional level of MAGL in TAMs cells. The constructed subcutaneous tumor model of mice demonstrated that up-regulation up-regulation of MAGL expression in M2-type TAMs inhibited the proliferation of MHCC97H human liver cancer cells in vivo. Furthermore, the mechanistic study illustrated that the high expression of MAGL promoted the transcription and secretion of inflammatory factors such as interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha in M2-type TAMs cells. Conclusion: The overexpression of MAGL inhibits the proliferation of MHCC97H hepatocellular carcinoma cells in vivo, and its mechanism may be associated to the release of inflammatory factors that from TAMs cells.
目的： 探讨单酰基甘油酯酶（MAGL）在M2型肿瘤相关巨噬细胞（TAMs）中不同表达对MHCC97H肝癌细胞体内增殖能力的影响及其机制。 方法： 将人外周血来源单核细胞诱导分化为M2型TAMs并通过流式细胞术鉴定。建立TAMs与MHCC97H肝癌细胞共培养模型，qRT-PCR检测TAMs细胞中MAGL表达。质粒转染MAGL在TAMs细胞中表达，qRT-PCR及酶联免疫吸附试验检测TAMs细胞的炎症因子mRNA水平及其分泌量。构建MHCC97H小鼠皮下荷瘤模型，观察MAGL在TAMs细胞中不同表达对MHCC97H肝癌细胞体内增殖的影响。计量资料的方差齐性检验用F检验，两独立样本均数之间的比较，方差齐者用t检验，方差不齐者用校正t检验。 结果： 成功诱导人外周血来源单核细胞分化为M2型TAMs。建立体外共培养模型，qRT-PCR检测发现MHCC97H肝癌细胞明显下调TAMs细胞中MAGL表达水平。构建小鼠皮下荷瘤模型，发现上调MAGL表达抑制M2型TAMs对MHCC97H肝癌细胞体内增殖的促进作用。进一步机制研究发现，MAGL高表达促进M2型TAMs细胞中炎症因子白细胞介素-1β、白细胞介素-6和肿瘤坏死因子-α的转录和分泌。 结论： MAGL高表达抑制MHCC97H肝癌细胞体内的增殖，其机制可能与增强TAMs细胞的炎症因子释放有关。.
Keywords: Carcinoma, hepatocellular; Inflammatory factors; M2-like TAMs; Monoacylglycerol lipase.