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SPARC accelerates biliary tract cancer progression through CTGF-mediated tumor–stroma interactions: SPARC as a prognostic marker of survival after neoadjuvant therapy

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Abstract

Purpose

In biliary tract cancer (BTC), malignancy is strongest at the invasion front. To improve the BTC prognosis, the invasion front should be controlled. We evaluated tumor–stroma crosstalk at the tumor center and at the invasion front of BTC lesions. We investigated the expression of SPARC, a marker of cancer-associated fibroblasts, and determined its ability to predict BTC prognosis after neoadjuvant chemoradiotherapy (NAC-RT).

Methods

We performed immunohistochemistry to evaluate SPARC expression in resected specimens from patients that underwent BTC surgery. We established highly invasive (HI) clones in two BTC cell lines (NOZ, CCLP1), and performed mRNA microarrays to compare gene expression in parental and HI cells.

Results

Among 92 specimens, stromal SPARC expression was higher at the invasion front than at the lesion center (p = 0.014). Among 50 specimens from patients treated with surgery alone, high stromal SPARC expression at the invasion front was associated with a poor prognosis (recurrence-free survival: p = 0.033; overall survival: p = 0.017). Coculturing fibroblasts with NOZ-HI cells upregulated fibroblast SPARC expression. mRNA microarrays showed that connective tissue growth factor (CTGF) was upregulated in NOZ-HI and CCLP1-HI cells. A CTGF knockdown suppressed cell invasion in NOZ-HI cells. Exogeneous CTGF upregulated SPARC expression in fibroblasts. SPARC expression at the invasion front was significantly lower after NAC-RT, compared to surgery alone (p = 0.003).

Conclusion

CTGF was associated with tumor–stroma crosstalk in BTC. CTGF activated stromal SPARC expression, which promoted tumor progression, particularly at the invasion front. SPARC expression at the invasion front after NAC-RT may serve as a prognosis predictor.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank San Francisco Edit (https://www.sfedit.net/) for the English language editing.

Funding

The authors declare that no funds, grants, or other support was received during the preparation of this manuscript.

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Authors and Affiliations

  1. Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Hirotoshi Takayama, Shogo Kobayashi, Kunihito Gotoh, Kazuki Sasaki, Yoshifumi Iwagami, Daisaku Yamada, Yoshito Tomimaru, Hirofumi Akita, Tadafumi Asaoka, Takehiro Noda, Hiroshi Wada, Hidenori Takahashi, Masahiro Tanemura, Yuichiro Doki & Hidetoshi Eguchi

  2. Department of Surgery, National Hospital Organization Osaka National Hospital, Osaka, Japan

    Kunihito Gotoh

  3. Department of Gastroenterological Surgery, Osaka International Cancer Institute, Osaka, Japan

    Hirofumi Akita & Hiroshi Wada

  4. Department of Surgery, Osaka Police Hospital, Osaka, Japan

    Tadafumi Asaoka

  5. Department of Surgery, Rinku General Medical Center, Osaka, Japan

    Masahiro Tanemura

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  1. Hirotoshi Takayama
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  2. Shogo Kobayashi
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Contributions

HT, SK, KG, YI and HE were involved in study design and data interpretation. HT, SK, KG, KS, YI, DY, YT, TN, HT, HA, HW, TA, MT, YD and HE were involved in the data analysis. All authors critically revised the report, commented on drafts of the manuscript, and approved the final report.

Corresponding author

Correspondence to Shogo Kobayashi.

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Conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Human Ethics Review Committee of the Graduate School of Medicine, Osaka University (No. 20493).

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Written informed consent was obtained from all patients included in the study.

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Not applicable as all data are anonymized.

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Supplementary Information

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432_2023_4835_MOESM1_ESM.tif

Supplementary fig. 1 SPARC is primarily expressed in fibroblast cells and rarely in biliary tract cancer (BTC) cells. (a) qRT-PCR results show SPARC mRNA expression and (b) Western blot shows SPARC protein expression in normal human dermal fibroblasts (NHDFs), BTC cells (NOZ-parental), and highly invasive BTC cells (NOZ-HI) (TIF 677 KB)

432_2023_4835_MOESM2_ESM.tif

Supplementary fig. 2 Evaluation of CTGF and SPARC immunostaining in BTC resected specimens. Immunohistochemistry of CTGF and SPARC in BTC resected specimens. (a) Representative figure of positive staining for CTGF (b) Representative figure of positive staining for SPARC (c) Representative figure of negative staining for CTGF (d) Representative figure of negative staining for SPARC. (a) and (b), (c) and (d) are the same field of the same specimen (TIF 8460 KB)

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Takayama, H., Kobayashi, S., Gotoh, K. et al. SPARC accelerates biliary tract cancer progression through CTGF-mediated tumor–stroma interactions: SPARC as a prognostic marker of survival after neoadjuvant therapy. J Cancer Res Clin Oncol 149, 10935–10950 (2023). https://doi.org/10.1007/s00432-023-04835-7

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