CHAF1B knockdown blocks migration in a hepatocellular carcinoma model

Peng,Xiaodong; Fu,Huiying; Yin,Jianjun; Zhao,Qingmei

doi:10.3892/or.2018.6437

CHAF1B knockdown blocks migration in a hepatocellular carcinoma model

Authors:
- Xiaodong Peng
- Huiying Fu
- Jianjun Yin
- Qingmei Zhao
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Affiliations: Department of Oncology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Oncology, The 94th Hospital of The People's Liberation Army, Nanchang, Jiangxi 330000, P.R. China, Department of Oncology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Oncology, Sichuan Mianyang 404 Hospital, Mianyang, Sichuan 361007, P.R. China
Published online on: May 16, 2018 https://doi.org/10.3892/or.2018.6437
Pages: 405-413

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Abstract

The roles and model of action of the chromatin assembly complex factor-1B (CHAF1B) gene in liver cancer have not been fully elucidated. The CHAF1B gene in human hepatocellular carcinoma cell line HUH-7 was knocked down using a lentivirus and the transfected cells were assayed for migration and invasion abilities and cell cycle arrest using the scratch wound healingand Transwell assays as well as flow cytometry, respectively. Cells transfected with an empty vector were used as the control. The expression of genes was profiled. Models were constructed using CHAF1B-knockdown cells and investigated for tumor growth and pathological changes. Our experiments revealed that the knockdown of the CHAF1 gene reduced the invasion and migration ability of HUH-7 cells. Gene expression profiling revealed that after knockdown, PSMB6, SLC30A7, SMC3, TWF2 and BLM genes had the most marked changes as compared with the control. Western blot and RT-PCR analyses revealed that following the knockdown of the CHAF1B gene, protein and mRNA levels of the PSMB6, SLC30A7 and SMC3 genes were significantly upregulated, while those of the BLM and TWF2 genes were significantly downregulated. In the HUH-7-knockdown cells, there were significantly fewer G0/G1 cells and more S1 cells as compared with the control (36.10 vs. 54.10% and 59.7 vs. 40.8%, respectively), while the number of G2/M cells was similar (4.20 vs. 5.10%). The volumes of the tumors were similar between those injected with the empty vector and control, but were significantly smaller in the knockdown models, suggesting that the knockdown of the CHAF1B gene inhibited tumor growth. H&E staining revealed that tumors were developed in mice in all groups.

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