Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Shenzhen University, Shenzhen Second People's Hospital, Shenzhen 518035, China.
Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China; Key Laboratory of Combined Multi-organ Transplantation, Ministry of Public Health, Hangzhou 310003, China.
Hepatobiliary Pancreat Dis Int. 2018 Aug;17(4):336-344. doi: 10.1016/j.hbpd.2018.07.006. Epub 2018 Jul 26.
Accumulating evidence demonstrates that microRNAs (miRNAs) play essential roles in tumorigenesis and cancer progression of hepatocellular carcinoma (HCC). Average targets of a miRNA were more than 100. And one miRNA may act in tumor via regulating several targets. The present study aimed to explore more potential targets of miR-449a by proteomics technology and further uncover the role of miR-449a in HCC tumorigenesis.
Technologies such as iTRAQ-based quantitative proteomic were used to investigate the effect of miR-449a on HCC. The expression of c-Met and miR-449a was detected by qRT-PCR in HCC samples. Gain- and loss-of-function experiments were performed to identify the function and potential target of miR-449a in HCC cells.
In HCC, miR-449a was significantly downregulated, while c-Met was upregulated concurrently. Quantitative proteomics and luciferase reporter assay identified c-Met as a direct target of miR-449a. Moreover, miR-449a inhibited HCC growth not only by targeting CDK6 but also by suppressing c-Met/Ras/Raf/ERK signaling pathway. Furthermore, the inhibition of c-Met expression with a specific siRNA significantly inhibited cells growth and deregulated the ERK pathway in HCC.
The tumor suppressor miR-449a suppresses HCC tumorigenesis by repressing the c-Met/ERK pathway.
越来越多的证据表明 microRNAs（miRNAs）在肝癌（HCC）的发生和发展中起着至关重要的作用。一个 miRNA 的平均靶标超过 100 个，而一个 miRNA 可能通过调节多个靶标在肿瘤中发挥作用。本研究旨在通过蛋白质组学技术探索 miR-449a 的更多潜在靶标，并进一步揭示 miR-449a 在 HCC 发生中的作用。
采用 iTRAQ 定量蛋白质组学等技术研究 miR-449a 对 HCC 的影响。通过 qRT-PCR 检测 HCC 样本中 c-Met 和 miR-449a 的表达。通过 gain-和 loss-of-function 实验，鉴定 miR-449a 在 HCC 细胞中的功能和潜在靶标。
在 HCC 中，miR-449a 明显下调，而 c-Met 则同时上调。定量蛋白质组学和荧光素酶报告基因实验鉴定 c-Met 是 miR-449a 的直接靶标。此外，miR-449a 不仅通过靶向 CDK6 抑制 HCC 生长，还通过抑制 c-Met/Ras/Raf/ERK 信号通路抑制 HCC 生长。此外，用特异性 siRNA 抑制 c-Met 表达显著抑制了 HCC 细胞的生长并使 ERK 通路失活。
抑癌 miR-449a 通过抑制 c-Met/ERK 通路抑制 HCC 肿瘤发生。
