miR⁃202及其靶基因Glypican⁃3在肝癌中的表达及其临床意义
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Expression and clinical significance of miR⁃202 and its target gene glypican⁃3 in hepatocellular carcinoma
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    摘要:

    目的:探究肝癌标志物磷脂酰肌醇蛋白聚糖3(glypican-3,GPC3)与miR-202相互调控的分子机制。方法:取126例临床肝癌组织标本及血清样本,免疫组化检测GPC3表达,qRT-PCR检测循环miR-202的水平;培养肝癌HepG2细胞,转染miR-202 mimics,qRT-PCR检测miR-202表达,Western blot检测GPC3表达;CCK8实验检测miR-202 mimics对HepG2细胞增殖活性的影响;Luciferase报告基因实验检测miR-202对GPC3的调控。结果:临床126例肝癌GPC3总阳性率为77.78%,其表达水平与患者性别、年龄、肿瘤大小、临床分期无关,但与细胞组织学分化类型以及微血管侵犯高度相关。肝癌患者循环miR-202呈低水平状态,且与癌组织GPC3表达水平呈负相关关系。上调HepG2细胞中miR-202表达,GPC3表达随之下调,且癌细胞增殖受抑制。Luciferase实验证实miR-202可直接负调控GPC3的表达。结论:GPC3受miR-202的直接调控,GPC3高表达与肝癌恶性生物学表征密切相关。靶向GPC3的治疗策略如能辅助提高miR-202的功能,将可能产生协同抗肝癌的效果。

    Abstract:

    Objective:This study aims to explore the molecular mechanism of the interaction between hepatocellular carcinoma(HCC) markers of glypican-3(GPC3)and miR-202. Methods:Total 126 cases of clinical HCC tissue samples and serum samples were collected. The expression of GPC3 was detected by immunohistochemistry,and the level of circulating miR-202 was detected by qRT-PCR. The HepG2 cells were cultured and transfected with miR-202 mimics,the expression of miR-202 was detected by qRT-PCR,and the expression of GPC3 protein was detected by Western blot. The effect of miR-202 mimics on the proliferation activity of HepG2 cells was detected by CCK8 assay. Luciferase reporter gene was used to determine the regulation of miR-202 on GPC3. Results:The total positive rate of GPC3 in 126 cases of HCC was 77.78%. Its expression level was not related to the patient’s gender,age,tumor size and clinical stage,but was highly correlated with the type of histological differentiation and microvascular invasion. The circulating miR-202 in HCC patients was at a low level,and was inversely correlated with the expression level of GPC3 in cancer tissues. The expression of miR-202 was up-regulated in HepG2 cells,and the expression of GPC3 was down-regulated accordingly,and the proliferation activity of cancer cells was thus inhibited. Luciferase experiment confirmed that miR-202 could directly and negatively regulate the expression of GPC3. Conclusion:This study demonstrates that GPC3 is directly regulated by miR-202,and high expression of GPC3 is a sign of malignant biological feature of HCC. Any targeted GPC3 therapeutic strategy,if it can assist to improve the function of miR-202,may produce a synergistic anti-tumor effect for HCC.

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周肖英,林 彬,侯云华,司淑平. miR⁃202及其靶基因Glypican⁃3在肝癌中的表达及其临床意义[J].南京医科大学学报(自然科学版),2021,(3):349-354

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  • 收稿日期:2020-05-13
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  • 在线发布日期: 2021-04-01
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