Tribbles假激酶3（）通过激活丝裂原活化蛋白激酶途径促进肝细胞癌的进展。
Tribbles pseudokinase 3 () contributes to the progression of hepatocellular carcinoma by activating the mitogen-activated protein kinase pathway.
作者信息
Wang Rui-Qi, He Fa-Zhong, Meng Qian, Lin Wei-Jie, Dong Jia-Mei, Yang Hai-Kui, Yang Yang, Zhao Min, Qiu Wen-Tao, Xin Yong-Jie, Zhou Zhi-Ling
机构信息
Department of Pharmacy, Zhuhai People's Hospital, Zhuhai Hospital Affiliated with Jinan University, Zhuhai, China.
Zhuhai Interventional Medical Center, Zhuhai Precision Medical Center, Zhuhai People's Hospital, Zhuhai Hospital Affiliated with Jinan University, Zhuhai, China.
出版信息
Ann Transl Med. 2021 Aug;9(15):1253. doi: 10.21037/atm-21-2820.
BACKGROUND
Tribble pseudokinase 3 () plays a key role in regulating the malignancy of many tumors. This study examined its function in cancer cells and explored the potential mechanisms of action.
METHODS
The expression of was examined in hepatocellular carcinomas (HCCs) using The Cancer Genome Atlas (TCGA) database. A lentivirus with a flag label was constructed and transfected into Huh7 and Hep3B human hepatoma cell lines to generate cells that stably overexpress . A small interfering RNA (siRNA) was designed to knockdown mRNA in HepG2 and Huh7. Cell viability and cell colony formation assays were conducted. Flow cytometry was performed to assess the cell cycle in cells overexpressing . Western blotting were performed to examine the expression of (Mitogen-activated protein kinase, MAPKK) (MEK), phosphorylated-MEK (p-MEK), extracellular signal-regulated kinase (ERK), and p-MEK in cells with knockdown. The correlation between and was assessed using co-immunoprecipitation assays and immunofluorescence.
RESULTS
was significantly overexpressed in advanced grade HCC tissues and was closely correlated with poor prognosis. overexpression promoted the cell growth and cell cycle but had little effect on migration capabilities in Huh7 and Hep3B cells. Conversely, knockdown of had slow down the cell growth in Huh7 and HepG2 cells detected by CCK8 and colony formation assay. The expression of and at both the protein and mRNA levels were downregulated when was knocked down. The protein expression of and were also downregulated upon silencing. is a transcript factor that is belongs to the SWI/SNF complex and has been shown to regulate many genes. Indeed, co-immunoprecipitation assays demonstrated that interacts with in the nucleus, suggesting that it may regulate in HCCs.
CONCLUSIONS
This study demonstrated that promotes the malignancy of HCC cells and its expression may be a potential diagnostic biomarker for HCC progression.
背景
Tribble假激酶3（）在调节多种肿瘤的恶性程度中起关键作用。本研究检测了其在癌细胞中的功能，并探讨了潜在的作用机制。
方法
使用癌症基因组图谱（TCGA）数据库检测肝细胞癌（HCC）中 的表达。构建带有flag标签的慢病毒并转染到Huh7和Hep3B人肝癌细胞系中，以产生稳定过表达 的细胞。设计小干扰RNA（siRNA）以敲低HepG2和Huh7中的 mRNA。进行细胞活力和细胞集落形成测定。进行流式细胞术以评估过表达 的细胞中的细胞周期。进行蛋白质印迹以检测敲低 的细胞中丝裂原活化蛋白激酶（MAPKK）（MEK）、磷酸化MEK（p-MEK）、细胞外信号调节激酶（ERK）和p-MEK的表达。使用免疫共沉淀测定和免疫荧光评估 与 的相关性。
结果
在高级别HCC组织中显著过表达，且与预后不良密切相关。 过表达促进Huh7和Hep3B细胞的细胞生长和细胞周期，但对迁移能力影响不大。相反，通过CCK8和集落形成测定检测到，敲低 会减缓Huh7和HepG2细胞的细胞生长。敲低 时， 和 在蛋白质和mRNA水平的表达均下调。沉默 后， 和 的蛋白质表达也下调。 是一种转录因子，属于SWI/SNF复合物，已被证明可调节许多基因。事实上，免疫共沉淀测定表明 在细胞核中与 相互作用，表明它可能在HCC中调节 。
结论
本研究表明， 促进HCC细胞的恶性程度，其表达可能是HCC进展的潜在诊断生物标志物。
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