Document Type : Original Article(s)


1 Department of Molecular Biology and Genetics, Haliç University, Ístanbul, Turkey

2 Department of Molecular Biology and Genetics, Atlas University, Ístanbul, Turkey



Background: Apoptosis and autophagy functions are known as death-inducing mechanisms. Vincristine (VCR) is a chemotherapeutic drug that binds with tubulin dimers and induces apoptosis in cancer cells. Furthermore, VCR protects cancer cells from apoptosis, allowing them to enter autophagy. This makes cancer even more aggressive in its latter stages. Herein, we aimed to investigate the effect of VCR on autophagy and apoptosis in gastric cancer AGS and MKN45 cells under hypoxic conditions.
Method: In this experimental approach, we performed cell culture, cell viability analysis, ribonucleic acid (RNA) isolation, cDNA synthesis, and observation of gene expression levels. Subsequently, the amount of acidic vesicular organelles was investigated with acridine orange staining. Through the use of quantitative real-time polymerase chain reaction, we studied the effect of hypoxia and VCR on mRNA levels of hypoxia-related vascular endothelium growth factor(VEGF) and hypoxia inducible factor-1a (HIF-1α), as well as apoptosis-related Bcl-2-associated X protein (Bax) and B-cell lymphoma 2 (Bcl-2), and autophagy-related Beclin-1 and microtubule-associated protein 1A/1B-light chain 3 (LC3-II) on AGS and MKN45 cell lines. The optimum VCR concentration was selected via cell viability analysis.
Results: The optimum VCR concentration was selected as 300 nM. VEGF, HIF- 1α, Bax, Bcl-2, and LC3-II mRNA expressions increased by 7.63, 3.34, 2.07, 4.09, and 3.1 folds, respectively, after VCR application to AGS cells under hypoxic conditions compared to the control cells. In MKN45 cells, Bax and LC3-II gene expressions increased by 5.21 and 12.2 folds, respectively, under the same conditions. In addition, the increase in the expression of autophagic LC3-II gene was consistent with acidic vesicular organelle formation.
Conclusion: It was observed that VCR affects hypoxic, apoptotic, and autophagic gene expressions in AGS and MKN45 cells under hypoxic conditions.


Meliha Burcu Irmak-Yazicioglu (PubMed)


How to cite this article:

Irmak- Yazicioglu MB, Ceylaner B. The effect of vincristine on hypoxic, apoptotic, and autophagic gene expressions in gastric cancer cells under hypoxic conditions. Middle East J Cancer. 2023;14(3):386-98. doi:10.30476/mejc.2023.94928.1750.

  1. Demetriou CA, Degli Esposti D, Pullen Fedinick K, Russo F, Robinson O, Vineis P. Filling the gap between chemical carcinogenesis and the hallmarks of cancer: A temporal perspective. Eur J Clin Invest. 2018;48(6): e12933. doi: 10.1111/eci.12933.
  2. Kankeu Fonkoua L, Yee NS. Molecular characterization of gastric carcinoma: therapeutic implications for biomarkers and targets. 2018;6(1):32. doi: 10.3390/biomedicines6010032.
  3. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007;35(4):495-516. doi: 10.1080/01926230701320337.
  4. Gültekin N, Karaoğlu K, Küçükateş E. New discoveries in the mechanisms of apoptosis and cell survival and novel potential therapeutic strategies. [In Turkish] Turk Kardiyol Dern Ars. 2008;36(2):120-30.
  5. Irmak MB, Ince G, Ozturk M, Cetin-Atalay R. Acquired tolerance of hepatocellular carcinoma cells to selenium deficiency: a selective survival mechanism? Cancer Res. 2003;63(20):6707-15.
  6. Fan XM, Wong BC, Wang WP, Zhou XM, Cho CH, Yuen ST, et al. Inhibition of proteasome function induced apoptosis in gastric cancer. Int J Cancer. 2001;93(4):481-8. doi: 10.1002/ijc.1373.
  7. Mizushima N, Yoshimori T, Ohsumi Y. The role of Atg proteins in autophagosome formation. Annu Rev Cell Dev Biol. 2011;27:107-32. doi: 10.1146/annurev-cellbio-092910-154005.
  8. Qian HR, Yang Y. Functional role of autophagy in gastric cancer. 2016;7(14):17641-51. doi: 10.18632/oncotarget.7508.
  9. Koukourakis MI, Giatromanolaki A, Sivridis E, Pitiakoudis M, Gatter KC, Harris AL. Beclin 1 over- and underexpression in colorectal cancer: distinct patterns relate to prognosis and tumour hypoxia. Br J Cancer. 2010;103(8):1209-14. doi: 10.1038/sj.bjc.6605904.
  10. Jiang ZF, Shao LJ, Wang WM, Yan XB, Liu RY. Decreased expression of Beclin-1 and LC3 in human lung cancer. Mol Biol Rep. 2012;39(1):259-67. doi: 10.1007/s11033-011-0734-1.
  11. Winship AL, Carpenter M, Griffiths M, Hutt KJ. Vincristine chemotherapy induces atresia of growing ovarian follicles in mice. Toxicol Sci. 2019;169(1):43-53. doi: 10.1093/toxsci/kfz022.
  12. Taghizadehghalehjoughi A, Sezen S, Hacimuftuoglu A, Güllüce M. Vincristine combination with Ca+2 channel blocker increase antitumor effects. Mol Biol Rep. 2019;46(2):2523-8. doi: 10.1007/s11033-019-04706-w.
  13. Høyer-Hansen M, Jäättelä M. Autophagy: an emerging target for cancer therapy. 2008;4(5):574-80. doi: 10.4161/auto.5921.
  14. Chen B, Jin F, Lu P, Lu XL, Wang PP, Liu YP, et al. Effect of mitogen-activated protein kinase signal transduction pathway on multidrug resistance induced by vincristine in gastric cancer cell line MGC803. World J Gastroenterol. 2004;10(6):795-9. doi: 10.3748/wjg.v10.i6.795.
  15. Hsich MJ, Hsieh YH, Lin CW, Chen MK, Yang SF, Chiou HL. Transcriptional regulation of Mcl-1 plays an important role of cellular protective effector of vincristine-triggered autophagy in oral cancer cells. Expert Opin Ther Targets. 2015;19(4):455-70. doi: 10.1517/14728222.2014.998200. Erratum in: Expert Opin Ther Targets. 2017;21(5):557-8.
  16. Yeo EJ, Chun YS, Park JW. New anticancer strategies targeting HIF-1. Biochem Pharmacol. 2004;68(6):1061-9. doi: 10.1016/j.bcp.2004.02.040.
  17. Mennerich D, Kubaichuk K, Kietzmann T. DUBs, hypoxia, and cancer. Trends Cancer. 2019;5(10):632-53. doi: 10.1016/j.trecan.2019.08.005.
  18. Fu JD, Yao JJ, Wang H, Cui WG, Leng J, Ding LY, et al. Effects of EGCG on proliferation and apoptosis of gastric cancer SGC7901 cells via down-regulation of HIF-1α and VEGF under a hypoxic state. Eur Rev Med Pharmacol Sci. 2019;23(1):155-61. doi: 10.26355/eurrev_201901_16759.
  19. Soni S, Padwad YS. HIF-1 in cancer therapy: two decade long story of a transcription factor. Acta Oncol. 2017;56(4):503-15. doi: 10.1080/0284186X.2017.1301680.
  20. Zhan Z, Li Q, Wu P, Ye Y, Tseng HY, Zhang L, et al. Autophagy-mediated HMGB1 release antagonizes apoptosis of gastric cancer cells induced by vincristine via transcriptional regulation of Mcl-1. 2012;8(1):109-21. doi: 10.4161/auto.8.1.18319.
  21. Park KJ, Yu MO, Park DH, Park JY, Chung YG, Kang SH. Role of vincristine in the inhibition of angiogenesis in glioblastoma. Neurol Res. 2016;38(10):871-9. doi: 10.1080/01616412.2016.1211231.
  22. Cheng J, Yang HL, Gu CJ, Liu YK, Shao J, Zhu R, et al. Melatonin restricts the viability and angiogenesis of vascular endothelial cells by suppressing HIF-1α/ROS/VEGF. Int J Mol Med. 2019;43(2):945-55. doi: 10.3892/ijmm.2018.4021.
  23. Zhang H, Bosch-Marce M, Shimoda LA, Tan YS, Baek JH, Wesley JB, et al. Mitochondrial autophagy is an HIF-1-dependent adaptive metabolic response to hypoxia. J Biol Chem. 2008;283(16):10892-903. doi: 10.1074/jbc.M800102200.
  24. Liu L, Ning X, Sun L, Zhang H, Shi Y, Guo C, et al. Hypoxia-inducible factor-1 alpha contributes to hypoxia-induced chemoresistance in gastric cancer. Cancer Sci. 2008;99(1):121-8. doi: 10.1111/j.1349-7006.2007.00643.x.
  25. Erler JT, Cawthorne CJ, Williams KJ, Koritzinsky M, Wouters BG, Wilson C, et al. Hypoxia-mediated down-regulation of Bid and Bax in tumors occurs via hypoxia-inducible factor 1-dependent and -independent mechanisms and contributes to drug resistance. Mol Cell Biol. 2004;24(7):2875-89. doi: 10.1128/MCB.24.7.2875-2889.2004.