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 Dhawan1 and Jagdeep Kaur1*
1Department of Biotechnology, Panjab University, Chandigarh, India
2Post Graduate Institute of Medical Education and Research, Chandigarh, India
*Corresponding Author: Jagdeep Kaur, Department of Biotechnology, Panjab University, Chandigarh, India.
Received:
January 09, 2023; Published: May 23, 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
- 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.
- 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.
- 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.
- 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.
- 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.
- Long J., et al. “Cancer statistics: current diagnosis and treatment of pancreatic cancer in Shanghai, China”. Cancer Letter2 (2014): 273.
- Ballinger A and Peikin SR. “Orlistat its current status as an anti-obesity drug”. European Journal of Pharmacology2-3 (2002): 109-117.
- 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.
- Kuhajda FPL. “Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology”. Nutrition 3 (2002): 202-208.
- 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.
- Menendez JA and Lupu R. “Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis”. Nature Reviews Cancer10 (2007): 763-777.
- Kuhajda FP. “Fatty acid synthase and cancer: new application of an old pathway”. Cancer Research12 (2006): 5977-5980.
- Kridel SJ., et al. “Orlistat is a novel inhibitor of fatty acid synthase with antitumor activity”. Cancer Research6 (2004): 2070-2075.
- Sadowski MC., et al. “The fatty acid synthase inhibitor triclosan: repurposing an anti-microbial agent for targeting prostate cancer”. Oncotarget19 (2014): 9362-9381.
- Sokolowska E., et al. “Orlistat reduces proliferation and enhances apoptosis in human pancreatic cancer cells (PANC-1)”. Anticancer Research11 (2017): 6321-6327.
- Codogno P., et al. “Autophagy and signaling: their role in cell survival and cell death”. Cell Death and Differentiation 2 (2015): 1509-1518.
- Yu L., et al. “The selectivity of autophagy and its role in cell death and survival”. Autophagy5 (2008): 567-573.
- 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.
- 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.
- Lin WC., et al. “Endoplasmic reticulum stress stimulates p53 expression through NF-κB activation”. PLoS One 7 (2012): e39120.
- Alirezaei M., et al. “Elevated ATG5 expression in autoimmune demyelination and multiple sclerosis”. Autophagy2 (2009): 152-158.
- 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.
- 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.
- 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.
- 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.
- Little JL., et al. “Inhibition of fatty acid synthase induces endoplasmic reticulum stress in tumor cells”. Cancer Research3 (2007): 1262-1269.
- 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.
- 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.
- 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.
- Raciti M., et al. “JNK2 is activated during ER stress and promotes cell survival”. Cell Death and Disease 11 (2012): e429.
- Rawla P., et al. “Epidemiology of Pancreatic Cancer: Global Trends, Etiology and Risk Factors”. World Journal of Oncology 1 (2019): 10-27.
- 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.
- Wright C., et al. “Anti-Tumorigenic Potential of a Novel Orlistat-AICAR Combination in Prostate Cancer Cells”. Journal of Cellular Biochemistry11 (2017): 3834-3845.
- Zhao L and Ackerman SL. “Endoplasmic reticulum stress in health and disease”. Current Opinion in Cell Biology 4 (2006): 444-452.
- Ogata M., et al. “Autophagy is activated for cell survival after endoplasmic reticulum stress”. Molecular and Cellular Biology24 (2006): 9220-9231.
- Ding WX., et al. “Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival”. Journal of Biological Chemistry 7 (2007): 4702-4710.
- 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.
- Zheng Y., et al. “Pinocembrin induces ER stress mediated apoptosis and suppresses autophagy in melanoma cells”. Cancer Letter 431 (2018): 31-42.
- 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.
- Shimizu S., et al. “Involvement of JNK in the regulation of autophagic cell death”. Oncogene 14 (2010): 2070-2082.
- Sui X., et al. “Autophagy and chemotherapy resistance: a promising therapeutic target for cancer treatment”. Cell Death and Disease10 (2013): e838.
- 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.
- Xu Y., et al. “Active autophagy in the tumor microenvironment: A novel mechanism for cancer metastasis”. Oncology Letter2 (2013): 411-416.
- 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.
- 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.
- Xu XD., et al. “Inhibition of Autophagy by Deguelin Sensitizes Pancreatic Cancer Cells to Doxorubicin”. International Journal of Molecular Sciences 2 (2017): 370.
Citation
Copyright