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Prognostic and Immunotherapeutic Predictive Value of CAD Gene in Hepatocellular Carcinoma: Integrated Bioinformatics and Experimental Analysis

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Abstract

Hepatocellular carcinoma (HCC) is a common type of cancer that ranks first in cancer-associated death worldwide. Carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase (CAD) are the key components of the pyrimidine pathway, which promotes cancer development. However, the function of CAD in HCC needs to be clarified. In this study, the clinical and transcriptome data of 424 TCGA-derived HCC cases were analyzed. The results demonstrated that high CAD expression was associated with poor prognosis in HCC patients. The effect of CAD on HCC was then investigated comprehensively using GO annotation analysis, KEGG enrichment analysis, Gene Set Enrichment Analysis (GSEA), and CIBERSORT algorithm. The results showed that CAD expression was correlated with immune checkpoint inhibitors and immune cell infiltration. In addition, low CAD levels in HCC patients predicted increased sensitivity to anti-CTLA4 and PD1, while HCC patients with high CAD expression exhibited high sensitivity to chemotherapeutic and molecular-targeted agents, including gemcitabine, paclitaxel, and sorafenib. Finally, the results from clinical sample suggested that CAD expression increased remarkably in HCC compared with non-cancerous tissues. Loss of function experiments demonstrated that CAD knockdown could significantly inhibit HCC cell growth and migration both in vitro and in vivo. Collectively, the results indicated that CAD is a potential oncogene during HCC metastasis and progression. Therefore, CAD is recommended as a candidate marker and target for HCC prediction and treatment.

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Funding

This study was supported by the National Natural Science Foundation of China (82072618), the National Key Research and Development Program of China (2022YFC2503700, 2022YFC2503705), and Shanghai Municipal Health Commission (2023ZZ02005).

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Author notes

  1. Xu Wang, Jin-Kai Feng, Fei-Fei Mao, and Yu-Chao Hou have contributed equally to this work.

Authors and Affiliations

  1. Cancer Center, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, 110 Ganhe Road, Shanghai, 200437, China

    Xu Wang, Yu-Chao Hou, Yu-Qing Zhang, Li-Heng Liu & Shu-Qun Cheng

  2. Department of Hepatic Surgery VI, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, 225 Changhai Road, Shanghai, 200433, China

    Xu Wang, Jin-Kai Feng, Yu-Chao Hou, Li-Heng Liu, Ju-Xian Sun, Chang Liu, Jie Shi & Shu-Qun Cheng

  3. Tongji University Cancer Center, School of Medicine, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, China

    Fei-Fei Mao & Shu-Qun Cheng

  4. The First Clinical Medicine School, Guangdong Pharmaceutical University, Guangzhou, China

    Qian Wei

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Contributions

Conception and design: S-QC, JS, XW, J-KF, F-FM, and Y-CH; Administrative support: S-QC and JS; Provision of study materials: S-QC; Study validation and data curation: XW, J-KF, F-FM, Y-CH, Y-QZ, L-HL, QW, J-XS, and CL; Data analysis and interpretation: XW, J-KF, and F-FM; Manuscript writing: All authors; Final approval of manuscript: All authors.

Corresponding authors

Correspondence to Jie Shi or Shu-Qun Cheng.

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The authors declare no conflicts of interest.

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The research protocol and acquirement of patients’ tumor samples were approved by the Institutional Ethics Committee of Eastern Hepatobiliary Surgery Hospital. All animal experiments were in agreement with the guidelines for the Care and Use of Laboratory Animals and were approved by the Ethics and Welfare Committee for Animal Research at the Yueyang Hospital of Integrated Traditional Chinese and Western Medicine.

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Wang, X., Feng, JK., Mao, FF. et al. Prognostic and Immunotherapeutic Predictive Value of CAD Gene in Hepatocellular Carcinoma: Integrated Bioinformatics and Experimental Analysis. Mol Biotechnol (2024). https://doi.org/10.1007/s12033-024-01125-6

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