Acta Scientific Pharmaceutical Sciences (ASPS)(ISSN: 2581-5423)

Research Article Volume 7 Issue 7

Orlistat Induced Endoplasmic Reticulum Stress Mediated Apoptosis and Protective Autophagy in PANC-1 Cells: The Key Role of JNK and Mitochondrial Dependent Signalling

Vishal Sharma1, Raja Ramachandran2, Samriti Dhawan3 and Jagdeep Kaur1*

1Department of Biotechnology, Panjab University, Chandigarh, India
2Post Graduate Institute of Medical Education and Research, Chandigarh, India
3Department of Biotechnology, Goswami Ganesh Dutta Sanatan Dharma College, Chandigarh, India

*Corresponding Author: Jagdeep Kaur, Department of Biotechnology, Panjab University, Chandigarh, India.

Received: January 09, 2023; Published: June 27, 2023

Abstract

Orlistat primarily designed to treat obesity is a lipase inhibitor, it is also reported to enhance apoptosis in Human Pancreatic Cancer Cells (PANC-1). Therefore efforts were made in the present investigation to evaluate the effect of orlistat on different parameters playing role in apoptosis and autophagy. Orlistat inhibited the PANC-1 cells growth in a dose-dependent manner with IC50 40 µM after 48 h. Treatment of PANC-1 with 10 µM and 20 µM orlistat for 48 h resulted in autophagy induction. Orlistat treatment of PANC-1 cells caused endoplasmic reticulum stress, as evidenced by increased cytosolic calcium levels, XBP1 splicing, GRP78, and CHOP up-regulation. Orlistat induced ROS generation and translocation of Bax from cytosol to mitochondrion with enhanced cytosolic cytochrome c level. Similarly, a simultaneously enhanced level of cytochrome c was found to be associated with caspase 3 activation and PARP cleavage. These observations suggested that orlistat induced endoplasmic reticulum stress, mitochondrion and ROS mediated cytotoxic action in PANC-1 cells. Further orlistat treatment reveals endoplasmic reticulum stress mediating autophagy through activation of the JNK pathway. To examine whether the autophagy induced by endoplasmic reticulum stress plays a role in cell survival or cell death, autophagy was blocked by 3-Methyladenine. Inhibition of orlistat induced autophagy using 3-Methyladenine results in enhanced apoptosis and suggested protective nature of orlistat induced autophagy in PANC-1. Collectively, all these studies suggested that orlistat had an anti-cancer effect on pancreatic cancer cells. In addition, autophagy played a pro-survival role, suppressing which the orlistat-induced anti-cancer effect would be more significant.

 Keywords: Orlistat; Pancreatic Cancer; Autophagy; Endoplasmic Reticulum Stress; Apoptosis

References

  1. Rahib L., et al. “Projecting cancer incidence and deaths to 2030: the unexpected burden of thyroid, liver, and pancreas cancers in the United States”. Cancer Research11 (2014): 2913-2921.
  2. Matrisian LM and Berlin JD. “The past, present, and future of pancreatic cancer clinical trials”. American Society of Clinical Oncology - Educational Book 35 (2016): e205-215.
  3. Souchek JJ., et al. “Combination treatment with orlistat-containing nanoparticles and taxanes is synergistic and enhances microtubule stability in taxane-resistant prostate cancer cells”. Molecular Cancer Therapeutics 9 (2017): 1819-1830.
  4. Chiou YS., et al. “Prevention of breast cancer by natural phytochemicals: focusing on molecular targets and combinational strategy”. Molecular Nutrition and Food Research23 (2018): e1800392.
  5. Qu CY., et al. “Engineering of lipid prodrug-based, hyaluronic acid-decorated nanostructured lipid carriers platform for 5-fluorouracil and cisplatin combination gastric cancer therapy”. International Journal of Nanomedicine 10 (2015): 3911-3920.
  6. Long J., et al. “Cancer statistics: current diagnosis and treatment of pancreatic cancer in Shanghai, China”. Cancer Letter2 (2014): 273.
  7. Ballinger A and Peikin SR. “Orlistat its current status as an anti-obesity drug”. European Journal of Pharmacology2-3 (2002): 109-117.
  8. Swierczynski J., et al. “Role of abnormal lipid metabolism in development, progression, diagnosis and therapy of pancreatic cancer”. World Journal of Gastroenterology9 (2014): 2279-2303.
  9. Kuhajda FPL. “Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology”. Nutrition 3 (2002): 202-208.
  10. Zaidi N., et al. “Lipogenesis and lipolysis: the pathways exploited by the cancer cells to acquire fatty acids”. Progress in Lipid Research 4 (2013): 585-589.
  11. Menendez JA and Lupu R. “Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis”. Nature Reviews Cancer10 (2007): 763-777.
  12. Kuhajda FP. “Fatty acid synthase and cancer: new application of an old pathway”. Cancer Research12 (2006): 5977-5980.
  13. Kridel SJ., et al. “Orlistat is a novel inhibitor of fatty acid synthase with antitumor activity”. Cancer Research6 (2004): 2070-2075.
  14. Sadowski MC., et al. “The fatty acid synthase inhibitor triclosan: repurposing an anti-microbial agent for targeting prostate cancer”. Oncotarget19 (2014): 9362-9381.
  15. Sokolowska E., et al. “Orlistat reduces proliferation and enhances apoptosis in human pancreatic cancer cells (PANC-1)”. Anticancer Research11 (2017): 6321-6327.
  16. Codogno P., et al. “Autophagy and signaling: their role in cell survival and cell death”. Cell Death and Differentiation 2 (2015): 1509-1518.
  17. Yu L., et al. “The selectivity of autophagy and its role in cell death and survival”. Autophagy5 (2008): 567-573.
  18. Liu D., et al. “Inhibition of autophagy by 3-MA potentiates cisplatin-induced apoptosis in esophageal squamous cell carcinoma cells”. Medical Oncology1 (2011): 105-101.
  19. Wang FM., et al. “Resveratrol triggers the pro-apoptotic endoplasmic reticulum stress response and represses pro-survival XBP1 signaling in human multiple myeloma cells”. Experimental Hematology10 (2011): 999-1006.
  20. Lin WC., et al. “Endoplasmic reticulum stress stimulates p53 expression through NF-κB activation”. PLoS One 7 (2012): e39120.
  21. Alirezaei M., et al. “Elevated ATG5 expression in autoimmune demyelination and multiple sclerosis”. Autophagy2 (2009): 152-158.
  22. Zhu S., et al. “Endoplasmic reticulum stress mediates aristolochic acid I-induced apoptosis in human renal proximal tubular epithelial cells”. Toxicology In Vitro5 (2012): 663-671.
  23. Livak KJ and Schmittgen TD. “Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C (T)) Method”. Methods 4 (2001): 402-408.
  24. Kumar A., et al. “A novel parthenin analog exhibits anti-cancer activity: activation of apoptotic signaling events through robust NO formation in human leukemia HL-60 cells”. Chemico-Biological Interactions3 (2011): 204-215.
  25. Watanabe Y., et al. “Interferon-gamma induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis”. Journal of Cellular Biochemistry2 (2003): 244-253.
  26. Little JL., et al. “Inhibition of fatty acid synthase induces endoplasmic reticulum stress in tumor cells”. Cancer Research3 (2007): 1262-1269.
  27. Yen YP., et al. “Arsenic induces apoptosis in myoblasts through a reactive oxygen species-induced endoplasmic reticulum stress and mitochondrial dysfunction pathway”. Archives of Toxicology6 (2012): 923-933.
  28. Liu ZW., et al. “Protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling pathway plays a major role in reactive oxygen species (ROS)-mediated endoplasmic reticulum stress-induced apoptosis in diabetic cardiomyopathy”. Cardiovascular Diabetology1 (2013): 1-6.
  29. Sharma V., et al. “Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling”. Cell Stress Chaperones3 (2015): 431-440.
  30. Raciti M., et al. “JNK2 is activated during ER stress and promotes cell survival”. Cell Death and Disease 11 (2012): e429.
  31. Rawla P., et al. “Epidemiology of Pancreatic Cancer: Global Trends, Etiology and Risk Factors”. World Journal of Oncology 1 (2019): 10-27.
  32. Wen X., et al. “Deconvoluting the role of reactive oxygen species and autophagy in human diseases”. Free Radical Biology and Medicine 65 (2013): 402-410.
  33. Wright C., et al. “Anti-Tumorigenic Potential of a Novel Orlistat-AICAR Combination in Prostate Cancer Cells”. Journal of Cellular Biochemistry11 (2017): 3834-3845.
  34. Zhao L and Ackerman SL. “Endoplasmic reticulum stress in health and disease”. Current Opinion in Cell Biology 4 (2006): 444-452.
  35. Ogata M., et al. “Autophagy is activated for cell survival after endoplasmic reticulum stress”. Molecular and Cellular Biology24 (2006): 9220-9231.
  36. Ding WX., et al. “Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival”. Journal of Biological Chemistry 7 (2007): 4702-4710.
  37. Cheng Y and Yang JM. “Survival and death of endoplasmic-reticulum-stressed cells: Role of autophagy”. World Journal of Biological Chemistry 10 (2011): 226-231.
  38. Zheng Y., et al. “Pinocembrin induces ER stress mediated apoptosis and suppresses autophagy in melanoma cells”. Cancer Letter 431 (2018): 31-42.
  39. Shen S., et al. “Blocking autophagy enhances the apoptotic effect of 18β-glycyrrhetinic acid on human sarcoma cells via endoplasmic reticulum stress and JNK activation”. Cell Death and Disease9 (2017): e3055.
  40. Shimizu S., et al. “Involvement of JNK in the regulation of autophagic cell death”. Oncogene 14 (2010): 2070-2082.
  41. Sui X., et al. “Autophagy and chemotherapy resistance: a promising therapeutic target for cancer treatment”. Cell Death and Disease10 (2013): e838.
  42. Sui X., et al. “p38 and JNK MAPK pathways control the balance of apoptosis and autophagy in response to chemotherapeutic agents”. Cancer Letter2 (2014): 174-179.
  43. Xu Y., et al. “Active autophagy in the tumor microenvironment: A novel mechanism for cancer metastasis”. Oncology Letter2 (2013): 411-416.
  44. Suzuki R., et al. “Genistein potentiates the antitumor effect of 5-Fluorouracil by inducing apoptosis and autophagy in human pancreatic cancer cells”. Anticancer Research9 (2014): 4685-4692.
  45. Talukdar S., et al. “MDA-9/Syntenin regulates protective autophagy in anoikis-resistant glioma stem cells”. Proceedings of the National Academy of Sciences of the United States of America 22 (2018): 5768-5773.
  46. Xu XD., et al. “Inhibition of Autophagy by Deguelin Sensitizes Pancreatic Cancer Cells to Doxorubicin”. International Journal of Molecular Sciences 2 (2017): 370.

Citation

Citation: Jagdeep Kaur., et al. “Orlistat Induced Endoplasmic Reticulum Stress Mediated Apoptosis and Protective Autophagy in PANC-1 Cells: The Key Role of JNK and Mitochondrial Dependent Signalling". Acta Scientific Pharmaceutical Sciences 7.7 (2023): 54-63.

Copyright

Copyright: © 2023 Jagdeep Kaur., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




Metrics

Acceptance rate32%
Acceptance to publication20-30 days

Indexed In




News and Events


  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for Upcoming Issue
    The last date for submission of articles for regular Issues is September 25, 2024.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of "Best Article of the Issue"
  • Welcoming Article Submission
    Acta Scientific delightfully welcomes active researchers for submission of articles towards the upcoming issue of respective journals.

Contact US