Digestive System Department, Affiliated Hospital of Guilin Medical College, Guilin 541001, Guangxi Province, China.
Digestive System Department, Affiliated Hospital of Guilin Medical College, Guilin 541001, Guangxi Province, China.
Asian Pac J Trop Med. 2015 Nov;8(11):923-929. doi: 10.1016/j.apjtm.2015.10.001. Epub 2015 Oct 9.
To investigate the expression and the regulation effect of cell growth of microRNA-577 in hepatocellular carcinoma (HCC).
qRT-PCR was applied to detect the relative expression of miR-577 in 70 paired HCC and matched tumor adjacent tissues collecting from resection between March 2011 and March 2014. Pearson chi-square test was used to analyze the relationship between the miR-577 expression and clinical features. The miR-577 mimics were transfected into HepG2 cells; cell cycles were detected by flow cytometry, cell proliferation was measured by MTT assay and BrdU incorporation assay, and cell apoptosis was determined by flow cytometry and caspase3/7 activity analysis. The expressions of β-catenin were measured by immunohistochemistry. Spearman correlation analysis was used to analyze the relationship between miR-577 and β-catenin. qRT-PCR and western-blot were used to detect the expression of β-catenin in transfected HepG2 cells.
The relative expressions of miR-577 was significantly lower in HCC tissues compared to the matched normal tumor-adjacent tissues (P < 0.05). Low expression of miR-577 was significantly associated with large tumor size (≥5 cm, P < 0.05) and advanced tumor node metastasis stage (III+IV, P < 0.05). Transfection of miR-577 mimics could inhibit repress cell proliferation, enhance cell apoptosis and block the cell cycles in G0/G1 phase (P < 0.05). miR-577 in HCC group had a significant negative correlation relationship with the expression of downstream target of β-catenin (P < 0.05). Both the mRNA and protein expression in HepG2 cells were down-regulated after transfection (P < 0.05).
Low expression of miR-577 is related to the malignant clinicopathological features in HCC tissues, and miR-577 may suppress HCC growth through down-regulating β-catenin.
研究微小 RNA-577 在肝细胞癌(HCC)中的表达及其对细胞生长的调控作用。
应用 qRT-PCR 检测 2011 年 3 月至 2014 年 3 月期间手术切除的 70 对 HCC 及其配对肿瘤旁组织中 miR-577 的相对表达。采用 Pearson 卡方检验分析 miR-577 表达与临床特征的关系。转染 miR-577 模拟物至 HepG2 细胞,流式细胞术检测细胞周期,MTT 比色法和 BrdU 掺入法检测细胞增殖,流式细胞术和 caspase3/7 活性分析检测细胞凋亡。免疫组织化学检测β-catenin 的表达。采用 Spearman 相关分析 miR-577 与β-catenin 之间的关系。qRT-PCR 和 Western blot 检测转染 HepG2 细胞中β-catenin 的表达。
与配对的正常肿瘤旁组织相比,HCC 组织中 miR-577 的相对表达明显降低(P<0.05)。miR-577 低表达与肿瘤体积较大(≥5cm,P<0.05)和肿瘤淋巴结转移分期较高(III+IV,P<0.05)显著相关。转染 miR-577 模拟物可抑制细胞增殖,增强细胞凋亡,阻滞细胞周期于 G0/G1 期(P<0.05)。miR-577 在 HCC 组与下游靶基因β-catenin 的表达呈显著负相关(P<0.05)。转染后 HepG2 细胞中 mRNA 和蛋白表达均下调(P<0.05)。
miR-577 的低表达与 HCC 组织中恶性临床病理特征相关,miR-577 可能通过下调β-catenin 抑制 HCC 生长。