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肿瘤与炎症密切相关,作者以往已证实内源性促炎症缓解介质脂氧素A4(Lipoxin A4,LXA4)能在整体和细胞水平发挥抗肝癌细胞增殖和转移的作用。为进一步探讨LXA4通过调节肿瘤相关巨噬细胞对肝癌细胞株HepG2 microRNAs(miRNAs)表达谱的影响,该文首先提取经脂多糖(LPS)或LPS+LXA4作用24 h的人巨噬细胞株U937培养上清液,分别称为ACM或LCM,以模拟肿瘤的炎症微环境,并用此上清液刺激HepG2细胞,24 h后提取细胞总RNA,采用microRNA芯片miRCURYTM LNA Array(V16.0)检测,计算各样本中的miRNAs标准值及比值。以两组间Hy3荧光标记信号强度的比值≤0.5或≥2为标准判定差异表达miRNA。Real-time PCR检测hsa-miR-623的相对含量以验证基因芯片的结果。结果发现,与对照组细胞相比,经过ACM作用24 h的HepG2细胞有35个miRNAs上调、130个miRNAs下调。LCM组与ACM组相比,HepG2细胞有185个miRNAs上调、71个miRNAs下调。Real-time PCR检测的结果证实,hsa-miR-623的变化与基因芯片趋势一致。综上所述,LXA4能通过肿瘤相关巨噬细胞而间接发挥其调节HepG2细胞miRNAs表达谱的作用。
Tumor and inflammation are closely related, the authors have previously demonstrated that lipoxin A4 (LXA4), an endogenous proinflammatory mediator, can exert anti-proliferative and metastatic effects on hepatocellular carcinoma cells both at the whole and at the cellular level. In order to further investigate the effect of LXA4 on the expression of HepG2 microRNAs (miRNAs) by regulating tumor-associated macrophages, we first cultured human macrophage cell line U937, which was treated with lipopolysaccharide (LPS) or LPS + LXA4 for 24 h The supernatants were respectively named as ACM or LCM to simulate the inflammatory microenvironment of the tumor. HepG2 cells were stimulated with this supernatant. The total RNA was extracted 24 h later and detected by microRNA chip miRCURYTM LNA Array (V16.0) MiRNAs in each sample standard values and ratios. Differentially expressed miRNAs were determined by the ratio of Hy3 fluorescence signal intensity between two groups ≤0.5 or ≥2. The relative content of hsa-miR-623 was detected by Real-time PCR to verify the results of the gene chip. The results showed that 35 miRNAs were up-regulated and 130 miRNAs were down-regulated in HepG2 cells treated with ACM for 24 h compared with control cells. In LCM group, 185 miRNAs were up-regulated and 71 miRNAs were down-regulated in HepG2 cells compared with ACM group. The result of Real-time PCR test confirmed that the change of hsa-miR-623 is consistent with the trend of gene chip. In summary, LXA4 can indirectly exert the regulation of miRNAs expression in HepG2 cells through tumor-associated macrophages.