SIRT3 Inhibits Cholangiocarcinoma Cell Proliferation by Targeting HIF1α to regulate the EMT Signaling Pathway

doi:10.21203/rs.3.rs-892577/v1

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SIRT3 Inhibits Cholangiocarcinoma Cell Proliferation by Targeting HIF1α to regulate the EMT Signaling Pathway

https://doi.org/10.21203/rs.3.rs-892577/v1

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Background

Cholangiocarcinoma (CCA), is a rare biliary adenocarcinoma associated with poor outcomes. Deacetylase Sirtuin‐3 (SIRT3), a histone deacetylase (HDAC), has been considered to be associated with various cancers and can be a potential new target for CCA.We intended to identify the target of SIRT3 and explore the mechanism of SIRT3 in CCA.

Methods

The expression levels of SIRT3 and hypoxia‐inducible factor‐1α (HIF1α) in CCA tissues and cell lines were examined by RT-qPCR. CCK-8 and EdU methods were used to detect cell proliferation in CCA. To assess the levels of proteins related to cell proliferation and epithelial-mesenchymal transition (EMT) process, western blot analysis was conducted. Co-Immunoprecipitation and deacetylation assays were used to explore HIF1α protein acetylation, stability and the relationship between SIRT3 and HIF1α in CCA cells.

Results

SIRT3 showed low expression in CCA tissues and cells. SIRT3 overexpression inhibited cell proliferation and EMT process. Moreover, the interaction between SIRT3 and HIF1α was confirmed and HIF1α expression was negatively regulated by SIRT3. Furthermore, we also found that HIF1α was more easily degraded and showed a reduction in stability through deacetylation via SIRT3 knockdown. In rescue assays, HIF1α also reversed the inhibitory effect of SIRT3 on cell proliferation and the EMT process.

Conclusions

SIRT3 suppressed cell proliferation and the EMT process in CCA by targeting HIF1α.

Trial registration

Samples were obtained only after the patient has given informed consent according to the established plan approved by the Ethics Committee of The First Affiliated Hospital of Anhui Medical University.

Oncology

Cholangiocarcinoma

SIRT3

HIF1α

EMT

Cholangiocarcinoma (CCA) is a primary liver malignancy, initially occurring in the intrahepatic biliary tract ¹. According to the existing statistic, the morbidity and mortality of CCA are increasing sharply each year due to its late clinical presentation, high conversion rate and poor prognosis ². CCA is often diagnosed at an advanced stage due to limited diagnostic methods ^{3, 4}. Surgery is the choice for most patients, however, because of the rapid progression and metastasis of CCA tumors, surgical treatment is not appropriate for all patients ⁵. In addition, radiotherapy or chemotherapy is still not ideal for improving the prognosis of patients with CCA ^{6, 7}. Thus, new therapeutic targets or strategies are urgently needed ⁸.

Sirtuin‐3 (SIRT3), is a nicotinamide adenine dinucleotide (NAD⁺)-dependent deacetylase in mitochondria ⁹. SIRT3 regulates cell survival, oxidative stress, apoptosis, metabolism, and through deacetylation ¹⁰. SIRT3 plays an important role in tumor promotion or inhibition in numerous cancers ^11-14. However, the possible biological role of SIRT3 in CCA and its potential molecular mechanism were not available in previous studies. Although previous study has reported the anti‐Warburg role of SIRT3 and has confirmed that SIRT3 inhibits the CCA progression by regulating the downstream pathway HIF1α/PDK1/PDHA1 ¹⁵. The Warburg effect represents the process of increased glucose uptake and aerobic glycolysis, which leads to the production of lactic acid, which becomes an early phenotype of cancer cells ^{16, 17}. The specific underlying regulatory mechanism of SIRT3 on CCA inhibition remains unclear.

Hypoxia inducible factor 1α, (HIF1α), has been found to regulate the expression of glycolytic‐related genes ¹⁸. Recently increasing evidence shows that HIF1α is involved in malignant disease progression ^{19, 20}. For example, increased HIF-1α levels are related to a poor prognosis in breast cancer ²¹. HIF1α protein was highly expressed in prostate, lung, and colon cancer ²². In cervical cancer, the deacetylation of HIF1α is increased by SIRT2, which reduces HIF1α degradation and increases HIF1α stability by further ubiquitination ²³. Thus, in this study, we hypothesized that HIF1α was modified by SIRT3 deacetylation to make it more susceptible to degradation and inhibit the proliferation of CCA. To test our hypothesis, we further explored the molecular mechanism and functional significance of SIRT3 to provide a novel research strategy for CCA treatment.

Cell culture

The CCA cell lines (HuCCT1, RBE and HCCC-9810) and a normal intrahepatic biliary epithelial cell (HIBEpiC) were used in this study (Chulabhorn Research Institute, Thailand). These CCA cell lines were maintained in RPMI-1640 supplemented with 10% FBS (Gibco, Grand Island, USA). A humidified incubator with 5% CO₂ was used to culture the cells at 37°C.

Cell transfection

pcDNA3.1 targeting SIRT3, HIF1α, and the negative control pcDNA3.1 were purchased from RiboBio, Guangzhou, China. The SIRT3 and HIF1α plasmids were from obtained from the Zhao Lab of Fudan University (Shanghai, China). CCA cells were transfected with pcDNA3.1-SIRT3, sh-SIRT3, and pcDNA3.1-HIF1α using the Lipofectamine RNAiMax reagent (Invitrogen).

Clinical specimens

Clinical tissue specimens of CCA from 20 patients were histopathologically diagnosed at The First Affiliated Hospital of Anhui Medical University (Hefei, China). Samples were obtained only after the patient has given informed consent according to the established plan approved by the Ethics Committee of The First Affiliated Hospital of Anhui Medical University.

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis

TRIzol reagent (Invitrogen, USA) was used to extract Total RNA from cells or tissues. Total RNA was reverse transcribed to cDNA by Reverse Transcription Kit (Takara, Dalian, China). Three replicates of each sample were amplified and analyzed with a Roche Light-Cycler (Roche, Basel, Switzerland). Results were normalized using GAPDH expression. The 2^−ΔΔCt method was used to calculate relative expression level. The primers used were as follows: SIRT3 forward, 5′-AGG GAC GAT TAT TAA AGG TGG A-3′ and reverse, 5′-TAC ATC CTG CAG GGA AAG C-3′; HIF1α forward, 5′-CCC ATT CCT CAC CCA TCA AAT A -3′ and reverse, 5′-CTT CTG GCT CAT ATC CCA TCA A-3′; GAPDH forward, 5′-GGG AAA CTG TGG CGT GAT-3′ and reverse, 5′-GAG TGG GTG TCG CTG TTG A-3′.

Cell counting Kit-8 (CCK-8) assay

CCA cells were seeded into 96‐well plates at the density of 2 × 10³ cells/well. Next, cells were cultured for 24, 48, or 72 h in an incubator with 5% CO₂. After 10 μL of CCK‐8 solution at 37°C for 2.5 h was added to each well, a microplate reader was applied to determine sample absorbance at 450 nm. The optical density (OD) value was measured, and the experiments were repeated three times.

5-Ethynyl-2′-deoxyuridine (EdU) assay

Cell proliferation was determined by EdU assay using a KeyFluor488 Click-iTEdU kit (KeyGEN, Nanjing, China) following the manufacturer’s instructions. About 5.0 × 10³ cells/well were seeded in a 96-well plate, and then transfected with pcDNA3.1-NC, pcDNA3.1-SIRT3, pcDNA3.1-SIRT3+pcDNA3.1-HIF1α. After transfection, EdU solution was added and the cells were incubated for 2 h at 37°C, and then fixed with 4% polyformaldehyde. Cells were then stained with DAPI. The cells were then visualized in three randomly chosen visual fields using the Zeiss fluorescence photomicroscope (Carl Zeiss, Oberkochen, Germany).

Western blot analysis

CCA cells were lysed with RIPA buffer (Beyotime, Shanghai, China). In order to separate the proteins from the sample buffer, 12% sodium dodecyl sulfate polyacrylamide (SDS–PAGE) gel was added, and then transferred onto a PVDF membrane, which was blocked with 5% nonfat milk for 1 h. The primary antibodies were provided by Abcam Company, including anti-SIRT3 (ab217319, 1:1000), anti-GAPDH (ab8245, 1:10000), anti- PCNA (ab92552, 1:1000), anti-Ki67 (ab15580, 1:1000), anti-CDK2 (ab32147, 1:1000), anti-E-cadherin (ab40772, 1:10000), anti-N-cadherin (ab76011, 1:5000), anti-Slug (ab27568, 1:1000), anti-Twist (ab50887, 1:1000), anti-HIF1α (ab270520, 1:1000) and anti-Tubulin (ab6160, 1:10000). The appropriate secondary antibodies were used at 1:2000 for all antibodies. Data were analyzed using ECL detection system (Pierce, Rockford, IL, USA).

Co-Immunoprecipitation assay

CCA cells were collected and dissolved in 500 μl co-IP buffer containing a a mixture of protease inhibitors (Sigma-Aldrich). Cells were prepared for extracting total proteins. Next, 1‐2 μg HIF1α antibody was added into 300 μL cell lysates at 4°C for 30 minutes. Then, proteins were incubated with the beads at 4°C overnight. On the following day, the beads were washed with for three times, and then, and the proteins were prepared for western blotting.

Deacetylation assay

HIF-1α and GFP proteins were expressed in HuCCT1 cells. In the deacetylation assay, the acetylated HIF-1α or SIRT3 immunoprecipitates were resuspended in 100 μl deacetylation buffer and 1 mM NAD. The Reactions were pretreated with 10 mM nicotinamide for 10 min and incubated for 2 h at 30 °C. Next, beads were placed on ice for 15 min and boiled with sample buffer. The supernatant was analyzed by Western blotting with anti-acetyllysine antibody.

Statistical analysis

Statistical analyses was performed with SPSS 19.0 software. The results presented as means ± standard deviation (SD). One-way analysis of variance (ANOVA) followed by LSD test was performed for multiple comparisons, and the student’s t test was used for comparisons between two groups The data were followed by Tukey’s post hoc test. p < 0.05 was considered statistically significant.

SIRT3 inhibits cell proliferation and EMT process

To investigate the role of SIRT3 in CCA, we first used RT-qPCR to analyze the expression of SIRT3 in 20 pairs of CCA tissues and 20 adjacent normal tissues, which showed that SIRT3 expression was lower in CCA tissues than in adjacent normal tissues (Fig. 1A). Decreased expression of SIRT3 was observed in HuCCT1, RBE, HCCC9810 cells, compared with HIBEpiC cells (Fig. 1B). Then we overexpressed SIRT3 and found elevated expression level of SIRT3 in HuCCT1 and RBE cells via RT-qPCR analysis (Fig. 1C). Declined cell viability induced by overexpressed SIRT3 was observed using a CCK-8 assay (Fig. 1D). The EdU incorporation assays also demonstrated that overexpression of SIRT3 reduced HuCCT1and RBE cell proliferation (Fig. 1E). Similarly, SIRT3 overexpression reduced the protein levels of PCNA, Ki67, CDK2, which were associated with cells proliferation in CCA cells (Fig. 1F). Western blotting also revealed that the SIRT3 overexpression increased E-cadherin protein level. On the contrary, proteins like N-cadherin, Slug, Twist with strong EMT capacity were suppressed (Fig. 1G). Based on the above findings, it can be concluded that SIRT3 was downregulated in cholangiocarcinoma, and SIRT3 overexpression inhibited cell proliferation and EMT process.

SIRT3 directly interacts with HIF1α in CCA cells

To explore the interaction between SIRT3 and HIF1α, co-immunoprecipitation (Co-IP) assays were performed. We found that SIRT3 and HIF1α precipitated each other as revealed by co-immunoprecipitation (Co-IP) assays, and the correlation between SIRT3 and HIF1α was initially identified (Fig. 2A). Protein level of HIF1α in lysed protein samples was observed, and we found that the protein level of HIF1α was reduced in SIRT3 overexpression group, while the protein level of HIF1α was increased in SIRT3 knockdown group in CCA cells (Fig. 2B). We analyzed the expression of HIF1α in 20 pairs of CCA tissues and 20 pairs of adjacent normal tissues by RT-qPCR, and the results showed that HIF1α was highly expressed in CCA tissues compared with adjacent normal tissues (Fig. 2C). The expression protein levels of HIF1α were also upregulated in CCA cell lines (Fig. 2D). Furthermore, HIF1α and SIRT3 expression showed a significant negative correlation in CCA tissues (Fig. 2E). Thus, these data suggested that SIRT3 contributed to proliferation by directly modulating HIF1α in CCA.

SIRT3 induces degradation of HIF1α via deacetylation

To further detect the relationship between SIRT3 and HIF1α, HIF1α was deacetylated by SIRT3 in HuCCT1 cells. SIRT3 knockdown showed high protein level of HIF1α as well as the HIF1α acetylation level, which indicated that SIRT3 induced HIF1α deacetylation (Fig. 3A). Then, we suppressed HIF1α synthesis by CHX (Cycloheximide) and HIF1α proteasomal degradation by MG132. In HuCCT1 cells, HIF1α degraded faster after SIRT3 overexpression, and the degradation of HIF1α was blocked by MG132 (Fig. 3B). Similarly, SIRT3 knockdown decreased the HIF1α-wt and HIF1α-mut protein stability, while sh-SIRT3 had no effect on the GFP and K709A (Fig. 3C). The results taken together indicated that HIF1α was more easily to be degraded and showed a reduction in stability through deacetylation via SIRT3 knockdown.

Overexpression of HIF1α reverses the inhibition of SIRT3 on cell proliferation and EMT process

To test the overexpression efficiency, we overexpressed HIF1α and found elevated expression levels of HIF1α in CCA HuCCT1 and RBE cells by RT-qPCR (Fig. 4A). The CCK-8 cell viability assay showed that HIF1α overexpression significantly reversed the inhibitory effects of SIRT3 overexpression on cell proliferation in CCA cells (Fig. 4B-C). EdU assay also showed that overexpression of HIF1α partially reversed the reduced proliferation of CCA cells induced by SIRT3 overexpression (Fig. 4D). The overexpression of HIF1α reversed the decreased levels of proliferation-related proteins (PCNA, Ki67, CDK2) induced by SIRT3 overexpression (Fig. 4E). In addition, western Blotting also showed that HIF1α overexpression reversed the promotive effect of SIRT3 overexpression on the protein and expression levels of E-cadherin, while HIF1α upregulation also reversed the suppressive effect of SIRT3 overexpression on the protein and expression levels of N-cadherin, Slug and Twist (Fig. 4F). These results further suggested that HIF1α overexpression reversed the inhibitory SIRT3 on cell proliferation and EMT process.

Sirtuin 3 (SIRT3) was found to have the most powerful mitochondrial deacetylase activity and is able to target key proteins for the normal function and metabolism of organelles ²⁴. Previous studies have found that SIRT3 acted as a tumor promoter in the development of some types of cancer, such as esophageal cancer ²⁵, breast cancer ²⁶, gastric cancer ²⁷, colon cancer ²⁸. On the contrary, SIRT3 has also been described as a tumor suppressor in some cancers ^{29, 30}. However, there are very few studies on SIRT3 in CCA. In our study, SIRT3 was lowly expressed in CCA tissues and cells. In addition, SIRT3 overexpression inhibited cell proliferation and EMT process. Next, we began to study the specific regulation mechanism of SIRT3 in CCA.

Previously, studies have shown that SIRT3 expression is closely associated with the expression of various HIF1α-dependent genes in breast cancer patients ³¹. In our study, we further confirmed the correlation between SIRT3 and HIF1α in CCA. HIF1α expression was negatively correlated with SIRT3 expression in CCA tissues, suggesting that SIRT3 contributed to cell proliferation by regulating HIF1α.

Acetylation is a key posttranslational modification and is also considered to be a key metabolic enzyme ^{32, 33}. Notably, in cervical cancer, SIRT2 increases HIF1α deacetylation, which reduces HIF1α degradation and increases HIF1α stability through further ubiquitination ²³. Deacetylases have also been investigated as potential therapeutic targets for CCA ³⁴. Our study confirmed that SIRT3 can indeed control glycolytic metabolism by modifying the stability and activity of HIF1α via deacetylation. We found that HIF1α modified with deacetylation was more easily degraded, which downregulated the expression level of HIF1α and decreased the stability of HIF1α. These results suggested that SIRT3 directly inhibited the function of HIF1α. Subsequently, we performed a rescue assay in which HIF1α overexpression reversed the suppressive effect of SIRT3 overexpression on cell proliferation and EMT process. Therefore, the decrease of HIF1α stability can enhance the inhibitory effect of SIRT3 on CCA cell proliferation and EMT process. In subsequent experiments, we will explore the effects of deacetylation and ubiquitination on the SIRT3/HIF1α axis and the mechanism of SIRT3 in various stages of tumor development.

In conclusion, SIRT3 exerted its inhibitory role on the proliferation and EMT process of CCA cells via regulating HIF1α. This finding may provide new research strategy for the CCA treatment.

Acknowledgement

We appreciate all participants who contributed to the study

Conflicts of interest

The authors declared no competing interests in this study.

Funding Sources

There is no funding to report for the research.

Ethics approval and consent to participate

Consent for publication

Not applicable.

Availability of data and materials

The datasets used during the current study are available from the corresponding author on reasonable request.

Authors' contributions

Yihang Zhao and Yaxian Kuai conceived and designed the experiments. Yihang Zhao, Yaxian Kuai, Jianhua Xu, Yufang Cui, Juan Wu, Mingming Zhang, Lei Xu, Lixing Zhou, Bin Sun and Yang Li carried out the experiments. Yihang Zhao, Yaxian Kuai, Bin Sun and Yang Li analyzed the data. Yihang Zhao, Yaxian Kuai, Bin Sun and Yang Li drafted the manuscript. All authors agreed to be accountable for all aspects of the work. All authors have read and approved the final manuscript.

1. Rizvi S and Gores GJ. Pathogenesis, diagnosis, and management of cholangiocarcinoma. Gastroenterology. 2013; 145: 1215-29.

2. Khan SA, Davidson BR, Goldin RD, et al. Guidelines for the diagnosis and treatment of cholangiocarcinoma: an update. Gut. 2012; 61: 1657-69.

3. Bridgewater J, Galle PR, Khan SA, et al. Guidelines for the diagnosis and management of intrahepatic cholangiocarcinoma. Journal of hepatology. 2014; 60: 1268-89.

4. Razumilava N and Gores GJ. Cholangiocarcinoma. Lancet (London, England). 2014; 383: 2168-79.

5. Skipworth JR, Olde Damink SW, Imber C, Bridgewater J, Pereira SP and Malagó M. Review article: surgical, neo-adjuvant and adjuvant management strategies in biliary tract cancer. Alimentary pharmacology & therapeutics. 2011; 34: 1063-78.

6. Torgeson A, Lloyd S, Boothe D, et al. Chemoradiation Therapy for Unresected Extrahepatic Cholangiocarcinoma: A Propensity Score-Matched Analysis. Annals of surgical oncology. 2017; 24: 4001-8.

7. Squadroni M, Tondulli L, Gatta G, Mosconi S, Beretta G and Labianca R. Cholangiocarcinoma. Critical reviews in oncology/hematology. 2017; 116: 11-31.

8. Rizvi S, Borad MJ, Patel T and Gores GJ. Cholangiocarcinoma: molecular pathways and therapeutic opportunities. Seminars in liver disease. 2014; 34: 456-64.

9. Alhazzazi TY, Kamarajan P, Verdin E and Kapila YL. SIRT3 and cancer: tumor promoter or suppressor? Biochimica et biophysica acta. 2011; 1816: 80-8.

10. Xiong Y, Wang M, Zhao J, Han Y and Jia L. Sirtuin 3: A Janus face in cancer (Review). International journal of oncology. 2016; 49: 2227-35.

11. Liu H, Li S, Liu X, Chen Y and Deng H. SIRT3 Overexpression Inhibits Growth of Kidney Tumor Cells and Enhances Mitochondrial Biogenesis. Journal of proteome research. 2018; 17: 3143-52.

12. Lee JJ, van de Ven RAH, Zaganjor E, et al. Inhibition of epithelial cell migration and Src/FAK signaling by SIRT3. Proceedings of the National Academy of Sciences of the United States of America. 2018; 115: 7057-62.

13. Wang S, Li J, Xie J, et al. Programmed death ligand 1 promotes lymph node metastasis and glucose metabolism in cervical cancer by activating integrin β4/SNAI1/SIRT3 signaling pathway. Oncogene. 2018; 37: 4164-80.

14. Xu LX, Hao LJ, Ma JQ, Liu JK and Hasim A. SIRT3 promotes the invasion and metastasis of cervical cancer cells by regulating fatty acid synthase. Molecular and cellular biochemistry. 2020; 464: 11-20.

15. Xu L, Li Y, Zhou L, et al. SIRT3 elicited an anti-Warburg effect through HIF1α/PDK1/PDHA1 to inhibit cholangiocarcinoma tumorigenesis. Cancer medicine. 2019; 8: 2380-91.

16. Vander Heiden MG, Cantley LC and Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science (New York, NY). 2009; 324: 1029-33.

17. Warburg O. On the origin of cancer cells. Science (New York, NY). 1956; 123: 309-14.

18. Finley LW, Carracedo A, Lee J, et al. SIRT3 opposes reprogramming of cancer cell metabolism through HIF1α destabilization. Cancer cell. 2011; 19: 416-28.

19. Ryan HE, Lo J and Johnson RS. HIF-1 alpha is required for solid tumor formation and embryonic vascularization. The EMBO journal. 1998; 17: 3005-15.

20. Talks KL, Turley H, Gatter KC, et al. The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumor-associated macrophages. The American journal of pathology. 2000; 157: 411-21.

21. Dales JP, Garcia S, Meunier-Carpentier S, et al. Overexpression of hypoxia-inducible factor HIF-1alpha predicts early relapse in breast cancer: retrospective study in a series of 745 patients. International journal of cancer. 2005; 116: 734-9.

22. Ravi R, Mookerjee B, Bhujwalla ZM, et al. Regulation of tumor angiogenesis by p53-induced degradation of hypoxia-inducible factor 1alpha. Genes & development. 2000; 14: 34-44.

23. Seo KS, Park JH, Heo JY, et al. SIRT2 regulates tumour hypoxia response by promoting HIF-1α hydroxylation. Oncogene. 2015; 34: 1354-62.

24. Verdin E, Hirschey MD, Finley LW and Haigis MC. Sirtuin regulation of mitochondria: energy production, apoptosis, and signaling. Trends in biochemical sciences. 2010; 35: 669-75.

25. Zhao Y, Yang H, Wang X, Zhang R, Wang C and Guo Z. Sirtuin-3 (SIRT3) expression is associated with overall survival in esophageal cancer. Annals of diagnostic pathology. 2013; 17: 483-5.

26. Torrens-Mas M, Pons DG, Sastre-Serra J, Oliver J and Roca P. SIRT3 Silencing Sensitizes Breast Cancer Cells to Cytotoxic Treatments Through an Increment in ROS Production. Journal of cellular biochemistry. 2017; 118: 397-406.

27. Cui Y, Qin L, Wu J, et al. SIRT3 Enhances Glycolysis and Proliferation in SIRT3-Expressing Gastric Cancer Cells. PloS one. 2015; 10: e0129834.

28. Liu C, Huang Z, Jiang H and Shi F. The sirtuin 3 expression profile is associated with pathological and clinical outcomes in colon cancer patients. BioMed research international. 2014; 2014: 871263.

29. Yu W, Denu RA, Krautkramer KA, et al. Loss of SIRT3 Provides Growth Advantage for B Cell Malignancies. The Journal of biological chemistry. 2016; 291: 3268-79.

30. Dong XC, Jing LM, Wang WX and Gao YX. Down-regulation of SIRT3 promotes ovarian carcinoma metastasis. Biochemical and biophysical research communications. 2016; 475: 245-50.

31. Haigis MC, Deng CX, Finley LW, Kim HS and Gius D. SIRT3 is a mitochondrial tumor suppressor: a scientific tale that connects aberrant cellular ROS, the Warburg effect, and carcinogenesis. Cancer research. 2012; 72: 2468-72.

32. Wang Q, Zhang Y, Yang C, et al. Acetylation of metabolic enzymes coordinates carbon source utilization and metabolic flux. Science (New York, NY). 2010; 327: 1004-7.

33. Zhao S, Xu W, Jiang W, et al. Regulation of cellular metabolism by protein lysine acetylation. Science (New York, NY). 2010; 327: 1000-4.

34. Pant K, Peixoto E, Richard S and Gradilone SA. Role of Histone Deacetylases in Carcinogenesis: Potential Role in Cholangiocarcinoma. Cells. 2020; 9.

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SIRT3 Inhibits Cholangiocarcinoma Cell Proliferation by Targeting HIF1α to regulate the EMT Signaling Pathway

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