Macrophages promote cell proliferation in colorectal cancer via IL-1β-mediated downregulation of miR-28-3p
Macrophages promote cell proliferation in colorectal cancer via IL-1β-mediated downregulation of miR-28-3p
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DOI：
10.23812/20-210-a
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发表时间：
2020-09-01
影响因子：
3.2
通讯作者：
Zheng, H-M
中科院分区：
文献类型：
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作者：
Fan, H-N;Liao, X-H;Zheng, H-M
巨噬细胞的浸润与多种恶性肿瘤的肿瘤进展和不良预后相关，但巨噬细胞导致结直肠癌（CRC）的潜在机制尚未阐明。本研究的目的是探讨巨噬细胞在结直肠癌中的作用机制。MTT法测定细胞活力。Western blot检测增殖相关标志物PCNA的表达。通过RNA干扰（RNAi）敲低巨噬细胞分泌的10种最重要的因子（PDGF、VEGF、TNF α、bFGF、IL-8、TGF-β、IFN-γ、IFN-γ、IL-1 β和IL-6），并通过qRT-PCR分析这10种因子的mRNA表达水平。MTT法检测上述因素对细胞增殖的影响。通过miRNA微阵列和qRT-PCR分析鉴定CRC细胞中受IL-1 β调控的miRNA。集落形成实验检测miR-28- 3 p抑制剂对大肠癌细胞增殖的影响。通过TCGA RNA-seq数据分析CRC患者中IL-1 β和miR-28- 3 p表达与临床病理特征的相关性。结果，巨噬细胞以时间和数量依赖性的方式促进CRC细胞的增殖，并且这些作用与PCNA和巨噬细胞分泌的细胞因子IL-1 β的上调相关，其对CRC细胞增殖具有最显著的作用。此外，在CRC细胞中，IL-1 β诱导miR-28- 3 p下调。miR-28- 3 p抑制剂促进CRC细胞的增殖。此外，IL-1 β表达的上调或miR-28- 3 p表达的下调与CRC患者的不良生存率相关。因此，这些数据表明巨噬细胞通过IL-1 β介导的miR-28- 3 p下调促进CRC细胞增殖。
Infiltration of macrophages is associated with tumor progression and poor prognosis in multiple malignancies, but the underlying mechanisms by which macrophages contribute to colorectal cancer (CRC) have not yet been elucidated. The purpose of this study was to discuss the potential mechanisms of macrophages in CRC. The MTT assay was used to assess cell viability. The expression of the proliferation-related marker PCNA was detected by Western blot analysis. The 10 most important factors (PDGF, VEGF, TNF alpha, bFGF, IL-8, TGF-beta, IFN-gamma, SPARC, IL-1 beta and IL-6) secreted by macrophages were knocked down by RNA interference (RNAi), and the mRNA expression levels of these 10 factors were analyzed by qRT-PCR. The effect of these factors on cell proliferation was assessed by the MTT assay. The miRNAs regulated by IL-1 beta in CRC cells were identified by miRNA microarray and qRT-PCR analyses. The proliferation ability of miR-28-3p inhibitor on CRC cells was detected by colony formation assay. The association of IL-1 beta and miR-28-3p expression with the clinicopathological characteristics in patients with CRC was analyzed by TCGA RNA-seq data. As a result, macrophages promoted the proliferation of CRC cells in a time- and number-dependent manner, and these effects were associated with the upregulation of PCNA and the macrophage-secreted cytokine IL-1 beta, which had the most significant effect on CRC cell proliferation. Furthermore, downregulation of miR-28-3p was induced by IL-1 beta in CRC cells. The miR-28-3p inhibitor promoted the proliferation in CRC cells. Moreover, upregulation of IL-1 beta expression or downregulation of miR-28-3p expression was associated with poor survival in patients with CRC. Therefore, these data demonstrated that macrophages promoted CRC cell proliferation via IL-1 beta-mediated downregulation of miR-28-3p.