ATOH8过表达抑制肝细胞癌的肿瘤进展和单核细胞趋化性。
ATOH8 overexpression inhibits the tumor progression and monocyte chemotaxis in hepatocellular carcinoma.
作者信息
Chen Liping, Yang Jingmao, Wang Yajie, Wu Nana, Li Xian, Li Jun, Huang Yangqing, Cheng Jilin
机构信息
Department of Gastroenterology, Shanghai Public Health Clinical Center, Fudan University Shanghai, China.
Department of Liver Surgery, Shanghai Public Health Clinical Center, Fudan University Shanghai, China.
出版信息
Int J Clin Exp Pathol. 2020 Oct 1;13(10):2534-2543. eCollection 2020.
OBJECTIVE
ATOH8 is reported to be associated with the progression of many tumors; however, there are remaining controversies. The aim of this study is to explore the role of ATOH8 in hepatocellular carcinoma (HCC) and its effect on monocyte chemotaxis.
METHODS
Bioinformatics analysis was performed based on the LIHC data in GEPIA and LinkedOmic. Fresh human liver cancer and adjacent nontumor tissue specimens were collected at the Shanghai Public Health Clinical Center. qRT-PCR was performed to determine the transcript level, and western blot analysis and ELISA were used to detect protein expression. CCK8, colony formation, wound-healing, Transwell migration and invasion assays were performed to examine cell proliferation, migration and invasion. An HCC xenograft mouse model was used to determine oncogenicity in vivo. Cell apoptosis and related markers were detected by flow cytometry. Additionally, chemotaxis was assessed by the Transwell migration assay.
RESULTS
ATOH8 expression is downregulated in HCC tissue and hepatoma cell lines. High expression of ATOH8 predicts a favorable prognosis. Overexpression of ATOH8 in liver cancer cells inhibits proliferation, migration and invasion in vitro, and tumor progression in nude mice. Knockdown of ATOH8 promotes proliferation of Huh7 and EMT-related proteins. Overexpression of ATOH8 increases chemosensitivity to 5-FU, which is probably caused by inhibiting the phosphorylation of AKT (Ser473). Furthermore, overexpression of ATOH8 in Huh7 reduced MCP1 to inhibit chemotactic THP-1, and promoted antitumor inflammatory cytokine (TNF-α and IFN-γ) secretion in monocytes.
CONCLUSION
In addition to the intrinsic oncosuppressive function of ATOH8 in the liver, ATOH8 may modulate the microenvironment to create an immune activation state. This may partly be attributed to ATOH8 inhibition of the monocyte recruitment via suppressing MCP1 expression so as to promote antitumor inflammatory cytokine secretion in monocytes.
目的
据报道，ATOH8与多种肿瘤的进展相关；然而，仍存在争议。本研究旨在探讨ATOH8在肝细胞癌（HCC）中的作用及其对单核细胞趋化性的影响。
方法
基于GEPIA和LinkedOmic中的LIHC数据进行生物信息学分析。在上海公共卫生临床中心收集新鲜的人类肝癌及癌旁非肿瘤组织标本。采用qRT-PCR测定转录水平，western blot分析和ELISA检测蛋白表达。进行CCK8、集落形成、伤口愈合、Transwell迁移和侵袭实验，以检测细胞增殖、迁移和侵袭能力。利用HCC异种移植小鼠模型确定体内致癌性。通过流式细胞术检测细胞凋亡及相关标志物。此外，通过Transwell迁移实验评估趋化性。
结果
ATOH8在HCC组织和肝癌细胞系中表达下调。ATOH8高表达预示着良好的预后。肝癌细胞中ATOH8的过表达抑制体外增殖、迁移和侵袭以及裸鼠体内肿瘤进展。敲低ATOH8促进Huh7细胞增殖及EMT相关蛋白表达。ATOH8的过表达增加对5-FU的化学敏感性，这可能是由于抑制AKT（Ser473）的磷酸化所致。此外，Huh7细胞中ATOH8的过表达降低MCP1以抑制THP-1的趋化，并促进单核细胞中抗肿瘤炎性细胞因子（TNF-α和IFN-γ）的分泌。
结论
除了ATOH8在肝脏中的内在抑癌功能外，ATOH8可能调节微环境以创造免疫激活状态。这可能部分归因于ATOH8通过抑制MCP1表达抑制单核细胞募集，从而促进单核细胞中抗肿瘤炎性细胞因子的分泌。
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