Radiosensitization Effect of Overexpression of Adenovirus-mediated SIRT6 on A549 Non-small Cell Lung Cancer Cells

  • Cai, Yong (Department of Radiation Oncology, Shanghai Pulmonary Hospital, Tongji University School of Medicine) ;
  • Sheng, Zhao-Ying (Department of Radiation Oncology, Shanghai Pulmonary Hospital, Tongji University School of Medicine) ;
  • Liang, Shi-Xiong (Department of Radiation Oncology, Shanghai Pulmonary Hospital, Tongji University School of Medicine)
  • Published : 2014.09.15


Objective: To explore the radiosensitization effect of overexpression of silent information regulator 6 (SIRT6) on A549 non-small cell lung cancer (NSCLC) cells. Methods: Adenovirus vector Ad-SIRT6 causing overexpression of SIRT6 was established. Western blotting and MTT assay were adopted to detect the level of SIRT6 protein and the inhibitory rate of A549 cell proliferation after different concentrations of adenovirus transduction (0, 25, 100, 200, and 400 pfu/cell) for 24 h. Control group, Ad-null group and Ad-SIRT6 group were designed in this experiment and virus concentration of the latter two groups was 200 pfu/cell. Colony formation assays were employed to test survival fraction (SF) of the 3 groups after 0, 2, 4, 6, 8, 10 X-ray irradiation. Flow cytometry was used to detect the status of cell cycle of 3 groups after 48 h of 4Gy X-ray irradiation and Western blotting was used to determine the expression of apoptosis-related genes of 3 groups after 48 h of 4GyX-ray irradiation. Results: In the range of 25~400 pfu/cell, the inhibitory rate of A549 cell proliferation increased as adenovirus concentration raised. The inhibitory rates under the concentrations of 0, 25, 100, 200, and 400 pfu/cell were 0%, $4.23{\pm}0.34%$, $12.7{\pm}2.57%$, $22.6{\pm}3.38%$, $32.2{\pm}3.22%$, $38.7{\pm}4.09%$ and $47.8{\pm}5.58%$ and there were significantly differences among groups (P<0.05). SF in Ad-SIRT6 group was lower than Ad-null and control groups after 4~10Gy X-ray irradiation (P<0.05) and the sensitization enhancement ratio (SER) was 1.35 when compared with control group. Moreover, after 48 h of 4Gy X-ray irradiation, there appeared a significant increase in G1-phase cell proportion, upregulated expression of the level of apoptosis-promoting genes (Bax and Cleaved caspase-3), but a obvious decline in S-phase and G2-phase cell proportion and a significant decrease of the level of apoptosis-inhibiting gene (Bal-2) in the Ad-SIRT6 group (P<0.05). Conclusion: The over-expression of adenovirus-mediated SIRT6, which has radiosensitization effect on A549 cells of NSCLC, can inhibit the proliferation of A549 cells and cause G0/G1 phase retardation as well as induce apoptosis of cells.


  1. Aydiner A, Ciftci R, Karabulut S, et al (2013). Does beta-blocker therapy improve the survival of patients with metastatic non-small cell lung cancer? Asian Pac J Cancer Prev, 14, 6109-14.
  2. Bonnet S, Archer SL, Allalunis-Turner J, et al (2007). A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell, 11, 37-51.
  3. Cai Y, Wang JY, Liu H (2013). Clinical observation of whole brain radiotherapy concomitant with targeted therapy for brain metastasis in non-small cell lung cancer patients with chemotherapy failure. Asian Pac J Cancer Prev, 14, 5699-703.
  4. Fantin VR, St-Pierre J, Leder P (2006). Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell, 9, 425-34.
  5. Finkel T, Deng CX, Mostoslavsky R (2009). Recent progress in the biology and physiology of sirtuins. Nature, 460, 587-91.
  6. Feng R, Guo JS, He JX, et al (2011). CMNa affect radiosentivivity and homogenegous adhesion of Tca-8113 cell line. China Oncol, 21, 17-21.
  7. Huang L, Wang RZ (2008). Cyclin and tumor radiosensitivity. J Xinjiang Med Univ, 31, 1510-2.
  8. Jemal A, Siegel R, Xu J, et al (2010). Cancer statistics. CA Cancer J Clin, 60, 277-300.
  9. Kaya V, Yildirim M, Demirpence O, et al (2013). Prognostic significance of basic laboratory methods in non- small-celllung cancer. Asian Pac J Cancer Prev, 14, 5473-6.
  10. Kim SH, Jang YW, Hwang P, et al (2011). The reno-protective effect of a phosphoinositide 3-kinase inhibitor wortmannin on streptozotocin-induced proteinuric renal disease rats. Exp Mol Med, 44, 45-51.
  11. Le A, Cooper CR, Gouw AM, et al (2010). Inhibition of lactate dehydrogenase A induces oxidative stress and inhibits tumor progression. Proc Natl Acad Sci USA, 107, 2037-42.
  12. Liu ZG, Yu ZC, Yu YP, et al (2010). Lysophosphatidic acid level and the incidence of silent brain infarction in patients with nonvalvular atrial fibrillation. Int J Mol Sci, 11, 3988-98.
  13. Ni XY, Qian N, Lin T, et al (2013). Radiosensitization effect of gold nanoparticles modified by sodium glycidazole on lung adenocarcioma cell A549. Chin J Radiol Med Protec, 33, 265-8.
  14. Natukula K, Jamil K, Pingali UR, et al (2013). The codon 399 Arg/Gln XRCC1 polymorphism is associated with lung cancer in Indians. Asian Pac J Cancer Prev, 14, 5275-9.
  15. Oven Ustaalioglu BB, Unal OU, Turan N, et al (2013). Prognostic factors for lymph node negative stage I and IIA non-small cell lung cancer: multicenter experiences. Asian Pac J Cancer Prev, 14, 6287-92.
  16. Sundaresan NR, Vasudevan P, Zhong L, et al (2012). The sirtuin SIRT6 blocks IGF-Akt signaling and development of cardiac hypertrophy by targeting c-Jun. Nat Med, 18, 1643-50.
  17. Sebastian C, Zwaans BM, Silberman DM, et al (2012). The histone deacetylase SIRT6 is a tumor suppressor that controls cancer metabolism. Cell, 151, 1185-99.
  18. Unal D, Eroglu C, Kurtul N, et al (2013). Are neutrophil/lymphocyte and platelet/lymphocyte rates in patients with non-small cell lung cancer associated with treatment response and prognosis? Asian Pac J Cancer Prev, 14, 5237-42.
  19. Vokes EE, Herndon JE II, J, Kelley MJ, et al (2007). Induction chemotherapy compared with chemoradiotherapy alone for regionally advanced unresectable stage III non-small-cell lung cancer: Cancer and Leukemia Group B. J Clin Oncol, 25, 1698-704.
  20. Vander Heiden MG, Cantley LC, Thompson CB (2009). Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science, 324, 1029-33.
  21. Wang JY, Cai Y (2013). Clinical observation and prognostic analysis of pemetrexed plus platinum as first-line treatment in patients with advanced non-small cell lung cancer. Asian Pac J Cancer Prev, 14, 6267-71.
  22. Wu GY, Hou PC, Wang W (2013). Radioresistance change and the machanism of human esophaged cancer EC706 cells in hypoxia. Chin J Radiol Med Protec, 33, 138-41.
  23. Zhong L, D'Urso A, Toiber D, et al (2010). The histone deacetylase Sirt6 regulates glucose homeostasis via Hif1alpha. Cell, 140, 280-93.

Cited by

  1. SIRTain regulators of premature senescence and accelerated aging vol.6, pp.5, 2015,
  2. Astragaloside IV sensitizes non–small cell lung cancer cells to gefitinib potentially via regulation of SIRT6 vol.39, pp.4, 2017,
  3. Low Rap1-interacting factor 1 and sirtuin 6 expression predict poor outcome in radiotherapy-treated Hodgkin lymphoma patients pp.1029-2403, 2017,