Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Mar 28;13(6):1985-2000.
doi: 10.7150/jca.69544. eCollection 2022.

ASF1b is a novel prognostic predictor associated with cell cycle signaling pathway in gastric cancer

Chuanzhi Chen  1 , Haili Bao  2 , Wu Lin  1 , Xiangliu Chen  1 , Yingying Huang  1 , Haohao Wang  1 , Yan Yang  1 , Jin Liu  1 , Xiadong Lv  1 , Lisong Teng  1
Affiliations

ASF1b is a novel prognostic predictor associated with cell cycle signaling pathway in gastric cancer

Chuanzhi Chen et al. J Cancer. .

Abstract

Gastric cancer (GC) is one of the most common malignant tumors with poor outcomes. Identification of new therapeutic targets is urgently needed. Accumulating evidence has shown that anti-silencing function 1b (ASF1b) contributes to the progression in multiple cancer types. However, detailed mechanisms of ASF1b tumorigenesis in gastric cancer remain elusive. This study showed that ASF1b was upregulated in GC tissues and remarkably correlated with TNM stage, histological grade and poor prognosis of GC. We induced down and up-regulation of ASF1b in GC cell lines and monitored the changes in their biological behavior. Furthermore, loss of ASF1b was efficient to suppress subcutaneous xenograft tumor growth in vivo. We demonstrate that ASF1b is involved in regulation of cell cycle and PI3K/AKT/mTOR signaling through experiments and database analysis. Mechanistically, ASF1b promoted the proliferation, migration and invasion of GC cells. Taken together, this study highlights the role of ASF1b, which provided new insights into the underlying mechanism of progression and metastasis in GC for the first time.

Keywords: ASF1b; Gastric cancer; PI3K/AKT/mTOR signaling.; cell cycle; cell proliferation; prognosis.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Figure 1
Figure 1
The integrative analytic strategy in this study. Stomach adenocarcinomas (STAD), Differential expression genes (DEGs), Kaplan-Meier Plotter database (KM-Plotter), Zhejiang University (ZJU).
Figure 2
Figure 2
(A,B) ASF1b mRNA levels of GC vs. normal adjacent tissues (NAT) in Oncomine database (Cho Gastric, DErrico Gastric ,P<0.0001). (C) GEPIA revealed ASF1b was significantly upregulated in 408 GC tissues. (D) ASF1b was significantly upregulated at most of tumors except Acute Myeloid Leukemia (LAML) and Testicular Germ Cell Tumors (TGCT) (E,F) The association between ASF1b expression and overall survival (OS), disease-free survival (DFS) in GC patients assessed by K-M plotter, respectively. (G,H) The association between ASF1b expression and OS, DFS in GC patients assessed by ZJU cohort, respectively. (I) Multivariable and univariable analyses were performed in Zhejiang cohort. All the bars correspond to 95% confidence intervals.
Figure 3
Figure 3
(A,B) ASF1b mRNA levels of GC vs. NAT in ZJU cohort. (B) Relative expression of ASF1b mRNA in GC tissues and their corresponding NAT was determined by qRT-PCR and expressed as -ΔΔCT (****P<0.0001). (C) Representative immunohistochemical staining of ASF1b in GC tissues and NAT (scale bar = 10 um). (D) A GC tissue microarray (TMA) (n = 160) indicated that the expression of ASF1b was upregulated in 65% of GC tissues. (E) The protein expression levels of ASF1b in 160 paired TMA (****P<0.0001).
Figure 4
Figure 4
(A,C) Protein expression and mRNA levels of ASF1b were checked in a panel of normal gastric cells (GES-1) and five human GC cell lines. (B,D) Protein expression and mRNA level of ASF1b were efficiently inhibited by ASF1b-si in AGS and MGC803 cells. (E,F) Migration assay was performed in AGS and MGC803 cells transfected with ASF1b-si/NC (scale bar = 20 μm, **P < 0.01; ***P < 0.001).
Figure 5
Figure 5
(A,B) Invasion assay was performed in AGS and MGC803 cells transfected with NC/ASF1b-si (scale bar = 20 μm, ***P < 0.001). (C,D) Colony formation assay in AGS and MGC803 cells transfected with NC/ASF1b-si. (E,F) Colony formation assay in AGS and MGC803 cells transfected with vector and ASF1b-overexpression (ASF1b-OE). (G) CCK-8 assay in AGS cells transfected with ASF1b-si/NC or vector/ASF1b-OE. (H) CCK-8 assay in MGC803 cells transfected with ASF1b-si/NC or vector/ASF1b-OE.
Figure 6
Figure 6
(A) KEGG pathway analysis of the genes significantly correlated with the ASF1b expression in GC from cBioPortal. Kyoto Encyclopedia of Genes and Genomes (KEGG). (B) GO analysis of ASF1b co-expressed genes in GC, Gene ontology (GO), Biological process(BP), Cellular component (CC), Molecular factor(MF). (C) MRNA levels of cell cycle-related markers in AGS cells transfected with ASF1b-si/NC using q-PCR. (D) MRNA levels of cell cycle-related markers in AGS cells transfected with vector/ASF1b-OE using q-PCR. (E) The protein expression levels of PI3K/AKT/mTOR pathway in GC cell lines after ASF1b-si. (F) The protein expression levels of cell cycle-related markers in GC cell lines after ASF1b-OE.
Figure 7
Figure 7
(A) Flow cytometry showing the percentages of AGS cells transfected with ASF1b-si/NC or vector/ASF1b-OE at different cell cycle phases. (B) Flow cytometry showing the percentages of MGC803 cells transfected with ASF1b-si/NC or vector/ASF1b-OE at different cell cycle phases.
Figure 8
Figure 8
(A) Protein expression and mRNA level of ASF1b were efficiently inhibited by Sh-ASF1b in AGS. (B) Representative images of subcutaneous tumors in BALB/c nude mice injected with AGS cells transferred with stably Sh-ASF1b/NC. (C,D) Tumor volumes and final body weights of mice injected with AGS cells transferred with stably Sh-ASF1b/NC. (E) ASF1b, Ki-67 and VEGF staining with corresponding antibody in xenografted AGS tumors after ASF1b silencing or NC, Vascular endothelial growth factor (VEGF), scale bar = 10 μm.
See this image and copyright information in PMC

References

    1. Ferlay J, Colombet M, Soerjomataram I, Parkin DM, Pineros M, Znaor A. et al. Cancer statistics for the year 2020: An overview. Int J Cancer. 2021;149:778–89. - PubMed
    1. Hamashima C. The burden of gastric cancer. Ann Transl Med. 2020;8:734. - PMC - PubMed
    1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A. et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71:209–49. - PubMed
    1. Collaborators GBDSC. The global, regional, and national burden of stomach cancer in 195 countries, 1990-2017: a systematic analysis for the Global Burden of Disease study 2017. Lancet Gastroenterol Hepatol. 2020;5:42–54. - PMC - PubMed
    1. Wei W, Zeng H, Zheng R, Zhang S, An L, Chen R. et al. Cancer registration in China and its role in cancer prevention and control. Lancet Oncol. 2020;21:e342–e9. - PubMed