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miR-577 通过靶向β-catenin 抑制肝癌细胞生长。

Inhibition effect of miR-577 on hepatocellular carcinoma cell growth via targeting β-catenin.

机构信息

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.

出版信息

Asian Pac J Trop Med. 2015 Nov;8(11):923-929. doi: 10.1016/j.apjtm.2015.10.001. Epub 2015 Oct 9.

Abstract

OBJECTIVE

To investigate the expression and the regulation effect of cell growth of microRNA-577 in hepatocellular carcinoma (HCC).

METHODS

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.

RESULTS

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).

CONCLUSIONS

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.

摘要

目的

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。

方法

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。

结果

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)。

结论

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。

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