Research paperTFAM depletion overcomes hepatocellular carcinoma resistance to doxorubicin and sorafenib through AMPK activation and mitochondrial dysfunction
Introduction
Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related deaths worldwide and the clinical management is still challenging (Chen et al., 2015). Most patients are diagnosed at the late stage when treatment is limited and ineffective. Chemo drug doxorubicin is used routinely for advanced HCC, and has shown inefficacy with a response rate of about ~20% (Cao et al., 2012). Sorafenib, the first approved tyrosine kinase inhibitor, prolongs median survival by 3 months over placebo (Spinzi and Paggi, 2008). Patients rapidly develop resistance to standard of care drugs with the underlying mechanisms largely unknown (Forner and Bruix, 2012). Accumulating evidence now suggests that metabolic deregulation via altered mitochondrial biogenesis and functions contributes to tumour progression and chemoresistance (Weinberg and Chandel, 2015).
Mitochondrial transcription factor A (TFAM), a nuclear-coded mitochondrial DNA packaging and transcription factor, is necessary for mitochondrial DNA (mtDNA) maintenance and mitochondrial biogenesis (Hallberg and Larsson, 2011). Studies in knockout mice have demonstrated that TFAM is required for functional oxidative phosphorylation and regulation of mtDNA copy number (Larsson et al., 1998). TFAM has been recently implicated in the involvement of tumor growth and invasion. Araujo and the colleagues report that TFAM shapes metabolic and invasion gene signatures in melanoma (Araujo et al., 2018). TFAM depletion leads to cell morphology change and growth-arrest in gastric cancer cells (Lee et al., 2017). In contrast, silencing of TFAM promotes intestinal tumorigenesis via a propensity for increased mitochondrial reactive oxygen species production and breast cancer growth (Woo et al., 2012, Balliet et al., 2011).
In this study, we investigated the expression pattern of TFAM in normal liver and HCC tissues and cells, sensitive and resistant HCC cell lines. We analysed the effect of TFAM depletion on HCC growth, survival, migration and drug resistance. Our findings demonstrate that TFAM is predominantly involved in the development of HCC resistance to standard of care treatment. TFAM depletion overcomes HCC resistance to doxorubicin and sorafenib through AMPK activation and mitochondrial dysfunction.
Section snippets
Immunohistochemical analysis and TFAM ELISA assay using cell and tissue samples
Paired tumor and normal liver tissues were obtained from treatment naïve-patients who underwent surgical treatment at Shenzhen Hospital of Southern Medical University. Written informed consent was obtained from all patients under institutional review board-approved protocols. The sectioned specimens were fixed with 4% paraformaldehyde (Sigma, USA). The samples were stained with anti-TFAM antibody and anti-Rabbit IgG, HRP-linked antibody, and DAB (3, 3-diaminobenzidine) according to
TFAM expression is upregulated in HCC response to anti-cancer drugs
To explore the expression pattern of TFAM in normal and malignant liver cells, we assessed TFAM protein level in paired normal liver and HCC tissues from HCC patients using both immunohistochemical and ELISA methods. Patients’ clinic-pathological features, including age, alpha fetoprotein (μg/l), tumor diameter, treatment and TNM stage, were presented in Supplementary Table 1. We found that TFAM expression is either moderately higher or similar in HCC tissues compared with corresponding normal
Discussion
TFAM activates mtDNA transcription, and affects nuclear gene expression through mitochondrial retrograde signaling. Although it is well known that TFAM is necessary for mtDNA maintenance and mitochondrial biogenesis, the role of TFAM in cancer is still inconclusive because both pro-cancer and anti-cancer effects are observed after TFAM depletion (Xie et al., 2016, Araujo et al., 2018, Lee et al., 2017a, Woo et al., 2012, Balliet et al., 2011). In this work, we report that 1) TFAM upregulation
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
This work was supported by research grants provided by Samming Project of Medicine in Shenzhen (No. ZHYX201904), National Natural Science Foundation of China (No. 81800503 and No. 81974070), The Basic Research Subject of Science and Technology plan in Bao’an District (No. 2019JD444), Natural Science Foundation of Liaoning Province (No. 201602758).
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These authors contributed to this work equally and are co-first authors.