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. 2023 Apr 27;25(6):256.
doi: 10.3892/ol.2023.13842. eCollection 2023 Jun.

Association between the expression levels of ADAMTS16 and BMP2 and tumor budding in hepatocellular carcinoma

Di Jiang  1 , Shaoshao Xu  1 , Chuanpeng Zhang  2 , Chuanbing Hu  3 , Lei Li  4 , Mingming Zhang  4 , Haiyan Wu  5 , Dongchang Yang  6 , Yanrong Liu  1   4
Affiliations

Association between the expression levels of ADAMTS16 and BMP2 and tumor budding in hepatocellular carcinoma

Di Jiang et al. Oncol Lett. .

Abstract

Tumor budding (TB) has become a crucial factor for predicting the malignancy grade and prognostic outcome for multiple types of solid cancer. Studies have investigated the prognostic value of TB in hepatocellular carcinoma (HCC). However, its molecular mechanism in HCC remains unclear. To the best of our knowledge, the present study was the first to compare the expression of differentially expressed genes (DEGs) between TB-positive (TB-pos) and TB-negative HCC tissues. In the present study, total RNA was extracted from 40 HCC tissue specimens and then sequenced. According to Gene Ontology (GO) functional annotation, upregulated DEGs were markedly associated with embryonic kidney development-related GO terms, which suggested that the TB process may at least partly mimic the process of embryonic kidney development. Subsequently, two genes, a disintegrin and metalloproteinase domain with thrombospondin motifs 16 (ADAMTS16) and bone morphogenetic protein 2 (BMP2), were screened and verified through immunohistochemical analysis of HCC tissue microarrays. According to the immunohistochemical results, ADAMTS16 and BMP2 were upregulated in TB-pos HCC samples, and BMP2 expression was increased in budding cells compared with the tumor center. Additionally, through cell culture experiments, it was demonstrated that ADAMTS16 and BMP2 may promote TB of liver cancer, thus promoting the malignant progression of liver cancer. Further analysis revealed that ADAMTS16 expression was associated with necrosis and cholestasis, and BMP2 expression was associated with the Barcelona Clinic Liver Cancer stage and the vessels encapsulating tumor clusters. Overall, the findings of the present study provided insights into the possible mechanisms of TB in HCC and revealed potential anti-HCC therapeutic targets.

Keywords: a metalloproteinase domain with thrombospondin motifs 16; bone morphogenetic protein 2; hepatocellular carcinoma; molecular mechanism; tumor budding.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1.
Figure 1.
Transcriptome sequencing of hepatocellular carcinoma tissues with different tumor budding statuses. (A) Volcano plot analysis of 95 upregulated DEGs and 150 downregulated DEGs. (B) Top 20 GO-BP terms for upregulated DEGs in TB-positive hepatocellular carcinoma tissues. (C) The top 20 GO-BP terms for downregulated DEGs in TB-positive hepatocellular carcinoma tissues. BPs, biological processes; DEGs, differentially expressed genes; FC, fold change; GO, Gene Ontology.
Figure 2.
Figure 2.
Immunohistochemical analysis of ADAMTS16 and BMP2 in patients with HCC with different TB statuses. (A) Representative IHC staining images of ADAMTS16 and BMP2 in TB-neg and TB-pos HCC tissues (magnification, ×100). Red boxes represent the tumor center and black arrows indicate budding tumor cells. (B) Representative IHC images of ADAMTS16 and BMP2 levels in the tumor center and budding cells (magnification, ×400), which are magnified views of the areas indicated by the red boxes and black arrows in (A), respectively. ADAMTS16, a metalloproteinase domain with thrombospondin motifs 16; BMP2, bone morphogenetic protein 2; HCC, hepatocellular carcinoma; IHC, immunohistochemistry; neg, negative; pos, positive; TB, tumor budding.
Figure 3.
Figure 3.
ADAMTS16 and BMP2 promote the TB of liver cancer in vitro. HepG2 cells were infected with ADAMTS16-OE lentivirus (GV513-ADAMTS16) or BMP2-OE lentivirus (GV358-BMP2) or corresponding control lentivirus (empty vector), CON335 and CON238, respectively. (A) Representative images (upper panel) and quantification of invaded cells (TB rate) (lower panel) in the inverse Matrigel invasion assays (magnification, ×100). The experiments were repeated independently three times. The data were analyzed using an unpaired Student's t-test. Quantitative results are presented as the mean ± SD. *P<0.05. (B) Representative images of the spheroid-based sprouting assays (magnification, ×200). Black arrows indicate budding HepG2 cells. ADAMTS16, a metalloproteinase domain with thrombospondin motifs 16; BMP2, bone morphogenetic protein 2; TB, tumor budding; OE, overexpression.
Figure 4.
Figure 4.
KM survival curves for ADAMTS16 and BMP2 expression. (A) KM curves (Cramer-von Mises test) for the overall survival of patients with HCC in the ADAMTS16 high-expression group (n=93) vs. the ADAMTS16 low-expression group (n=149). (B) KM curves (Cramer-von Mises test) for overall survival of patients with HCC in the BMP2 high-expression group (n=82) vs. the BMP2 low-expression group (n=160). (C) KM curves (log-rank test) for overall survival of patients with HCC with high/low ADAMTS16 expression using The Cancer Genome Atlas data. (D) KM curves (log-rank test) for overall survival of patients with HCC with high/low BMP2 expression using The Cancer Genome Atlas data. (E) KM curves (log-rank test) for overall survival of patients with HCC with high/low ADAMTS16 expression using the GSE76427 dataset. (F) KM curves (Cramer-von Mises test) for overall survival of patients with HCC with high/low BMP2 expression. ADAMTS16, a metalloproteinase domain with thrombospondin motifs 16; BMP2, bone morphogenetic protein 2; HCC, hepatocellular carcinoma; KM, Kaplan-Meier; TPM, transcripts per million.
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References

    1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021;71:209–249. doi: 10.3322/caac.21660. - DOI - PubMed
    1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424. doi: 10.3322/caac.21492. - DOI - PubMed
    1. Yang JD, Hainaut P, Gores GJ, Amadou A, Plymoth A, Roberts LR. A global view of hepatocellular carcinoma: Trends, risk, prevention and management. Nat Rev Gastroenterol Hepatol. 2019;16:589–604. doi: 10.1038/s41575-019-0186-y. - DOI - PMC - PubMed
    1. Turajlic S, Swanton C. Metastasis as an evolutionary process. Science. 2016;352:169–175. doi: 10.1126/science.aaf2784. - DOI - PubMed
    1. Hase K, Shatney C, Johnson D, Trollope M, Vierra M. Prognostic value of tumor ‘budding’ in patients with colorectal cancer. Dis Colon Rectum. 1993;36:627–635. doi: 10.1007/BF02238588. - DOI - PubMed

Grants and funding

The present study was funded by the National Natural Science Foundation of China (grant no. 81972629), the Taishan Scholars Program of Shandong Province (grant no. tsqn201909193), the Shandong Youth Innovation and Technology program (grant no. 2020KJL003), the Jining Research and Development Program (grant nos. 2021YXNS065 and 2021YXNS075), and the Research Fund for Lin He Academician New Medicine (grant no. JYHL2021FMS12).