Role of SIRT-3, p-mTOR and HIF-1α in Hepatocellular Carcinoma Patients Affected by Metabolic Dysfunctions and in Chronic Treatment with Metformin
Abstract
:1. Introduction
2. Results
2.1. Patient Characteristics
2.2. Immunohistochemical Expression Pattern of SIRT-3, p-mTOR and Nuclear HIF-1α in Tumor Tissues
2.3. In Vitro Effect of Metformin and Sorafenib on HCC Cell Lines
2.4. Association between SIRT-3, p-mTOR and Nuclear HIF-1α Expression and Clinical Outcome of Early-Stage HCC Patients
2.5. High p-mTOR Levels Correlated with Poor Prognosis in Advanced-Stage HCC Patients
3. Discussion
4. Materials and Methods
4.1. Patient Population
4.2. Immunohistochemistry Analysis
4.3. HCC Cell Lines and Drugs
4.4. Cell Viability Assay
4.5. Annexin V Assay
4.6. Western Blot Analysis
4.7. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
HCC | hepatocellular carcinoma |
NASH | non-alcoholic steatohepatitis |
SIRT-3 | sirtuin-3 |
T2DM | type 2 diabetes mellitus |
mTOR | mammalian target of rapamycin |
AMPK | AMP-activated protein kinase |
HIF-1α | hypoxia-induced factor-1α |
AASLD | American Association for the Study of Liver Diseases |
IQR | interquartile range |
DFS | disease-free survival |
PFS | progression-free survival |
OS | overall survival |
FBS | fetal bovine serum |
MTT | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
DMSO | dimethyl sulfoxide |
IR | insulin resistance |
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Patients | N. (%) |
---|---|
Median age (range) | 70 (28–89) |
Gender | |
Male | 60 (85.7) |
Female | 10 (14.3) |
Etiology | |
Viral infection | 31 (44.3) |
Metabolic syndrome | 23 (32.9) |
Alcoholic | 7 (10.0) |
Other | 9 (8.6) |
Diabetes | |
No | 38 (55.1) |
Yes | 31 (44.9) |
Metformin | |
No | 46 (66.7) |
Yes | 23 (33.3) |
Performance Status (ECOG) | |
0 | 56 (80) |
1 | 14 (20) |
Child–Pugh | |
A | 67 (89.7) |
B | 3 (10.3) |
BCLC staging | |
A | 41 (58.6) |
B | 9 (12.9) |
C | 20 (28.5) |
Alfafetoprotein levels | |
<400 | 17 (63.0) |
≥400 | 10 (37.0) |
Unknown | 43 |
MELD score | |
≤10 | 18 (25.7) |
>10 | 52 (74.3) |
Metastasis | |
No | 56 (80) |
Yes | 14 (20) |
Markers | No. Patients | Five-Year Percent DFS (months) (95% CI) | p | Eight-Year Percent OS (months) (95% CI) | p |
---|---|---|---|---|---|
SIRT-3 (%) | |||||
<35 | 23 | 23 (4–41) | 20 (0–49) | ||
≥35 | 23 | 51 (28–73) | 0.308 | 32 (0–78) | 0.117 |
p-mTor (%) | |||||
0 | 23 | 47 (22–73) | 39 (0–95) | ||
>0 | 4 | 50 (1–99) | 0.952 | 0 | 0.577 |
HIF-1α (%) | |||||
<1 | 15 | 20 (0–44) | 0 | ||
≥1 | 10 | 100 | 0.002 | 50 (0-100) | 0.041 |
Response | No. Patients | SIRT-3 (%) Median Value (range) | p | No. Patients | p-mTOR (%) Median value (range) | p | No. Patients | HIF-1α (%) Median Value (range) | p |
---|---|---|---|---|---|---|---|---|---|
CR | 4 | 80 (60–100) | 2 | 0 (0–0) | 2 | 20 (0–40) | |||
SD | 10 | 45 (0–100) | 7 | 0 (0–0) | 7 | 0 (0–8) | |||
PD | 10 | 55 (0–90) | 0.276 | 5 | 0 (0–80) | 0.407 | 4 | 0 (0–60) | 0.752 |
Markers | No. Patients | Median PFS (months) (95% CI) | p | Median OS (months) (95% CI) | p |
---|---|---|---|---|---|
SIRT-3 (%) | |||||
<60 | 11 | 3.7 (1.6–6.0) | 12.0 (5.2–nr) | ||
≥60 | 10 | 5.3 (1.7–12.9) | 0.542 | 14.3 (2.0–15.8) | 0.624 |
p-mTor (%) | |||||
0 | 13 | 5.3 (2.3–10.7) | 13.9 (6.7–15.8) | ||
>0 | 3 | 1.8 (1.6–4.0) | 0.055 | 6.1 (2.6–nr) | 0.098 |
HIF-1α (%) | |||||
0 | 10 | 3.5 (1.2–6.0) | 13.9 (2.2–15.8) | ||
>0 | 6 | 4.7 (1.6–22.8) | 0.382 | 12.0 (2.6–22.8) | 0.682 |
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De Matteis, S.; Scarpi, E.; Granato, A.M.; Vespasiani-Gentilucci, U.; La Barba, G.; Foschi, F.G.; Bandini, E.; Ghetti, M.; Marisi, G.; Cravero, P.; et al. Role of SIRT-3, p-mTOR and HIF-1α in Hepatocellular Carcinoma Patients Affected by Metabolic Dysfunctions and in Chronic Treatment with Metformin. Int. J. Mol. Sci. 2019, 20, 1503. https://doi.org/10.3390/ijms20061503
De Matteis S, Scarpi E, Granato AM, Vespasiani-Gentilucci U, La Barba G, Foschi FG, Bandini E, Ghetti M, Marisi G, Cravero P, et al. Role of SIRT-3, p-mTOR and HIF-1α in Hepatocellular Carcinoma Patients Affected by Metabolic Dysfunctions and in Chronic Treatment with Metformin. International Journal of Molecular Sciences. 2019; 20(6):1503. https://doi.org/10.3390/ijms20061503
Chicago/Turabian StyleDe Matteis, Serena, Emanuela Scarpi, Anna Maria Granato, Umberto Vespasiani-Gentilucci, Giuliano La Barba, Francesco Giuseppe Foschi, Erika Bandini, Martina Ghetti, Giorgia Marisi, Paola Cravero, and et al. 2019. "Role of SIRT-3, p-mTOR and HIF-1α in Hepatocellular Carcinoma Patients Affected by Metabolic Dysfunctions and in Chronic Treatment with Metformin" International Journal of Molecular Sciences 20, no. 6: 1503. https://doi.org/10.3390/ijms20061503