Skip to main content
Top
Published in: Cancer Cell International 1/2013

Open Access 01-12-2013 | Primary research

Beclin1 inhibition promotes autophagy and decreases gemcitabine–induced apoptosis in Miapaca2 pancreatic cancer cells

Authors: Xiaoshu Li, Jun Yan, Lisheng Wang, Fengjun Xiao, Yuefeng Yang, Xiaozhong Guo, Hua Wang

Published in: Cancer Cell International | Issue 1/2013

Login to get access

Abstract

Background

Beclin1 is a well-known key regulator of autophagy, which is also a haploinsufficient tumor suppressor. Current studies revealed that down-regulation or monoallelic deletions of Beclin1 were frequently found in various cancers. The purpose of this study was to investigate the effects of Beclin1 inhibition on autophagy and Gemcitabine-induced apoptosis of pancreatic cancer cells.

Methods

Beclin1 expression was inhibited by siRNA transduction and gene expression was determined by Real-time PCR and Western blot. The effects of Beclin1 inhibition on autophagy and Gemcitabine-induced apoptosis of Miapaca2 cells were analyed through LC3 expression, cell viability, cell cycle and apoptosis by using Western blot.

Results

We observed that Beclin1 silence promoted microtubule-associated protein 1 light chain 3-II (LC3-II) protein formation and increased punctate fluorescent signals in Miapaca2 cells transfected with green fluorescent protein (GFP)-tagged LC3. Beclin1 inhibition showed a greater suppressive effect on Gemcitabine-induced apoptosis of Miapaca2 cells.

Conclusion

Our data suggested that Beclin1 silence not only up-adjusted autophagy process, but also played an important role in the regulation of apoptosis. Beclin1 inhibition could inhibit apoptosis signaling induced by Gemcitabine in Miapaca2 cells.
Appendix
Available only for authorised users
Literature
1.
go back to reference Mizushima N, Levine B, Cuervo AM, Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 2008, 451 (7182): 1069-1075. 10.1038/nature06639.PubMedCentralCrossRefPubMed Mizushima N, Levine B, Cuervo AM, Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 2008, 451 (7182): 1069-1075. 10.1038/nature06639.PubMedCentralCrossRefPubMed
3.
go back to reference Brech A, Ahlquist T, Lothe RA, Stenmark H: Autophagy in tumour suppression and promotion. Mol Oncol. 2009, 3 (4): 366-375. 10.1016/j.molonc.2009.05.007.CrossRefPubMed Brech A, Ahlquist T, Lothe RA, Stenmark H: Autophagy in tumour suppression and promotion. Mol Oncol. 2009, 3 (4): 366-375. 10.1016/j.molonc.2009.05.007.CrossRefPubMed
4.
go back to reference Levine B, Yuan J: Autophagy in cell death: an innocent convict?. Clin Invest. 2005, 115 (10): 2679-2688. 10.1172/JCI26390.CrossRef Levine B, Yuan J: Autophagy in cell death: an innocent convict?. Clin Invest. 2005, 115 (10): 2679-2688. 10.1172/JCI26390.CrossRef
5.
go back to reference Katayama M, Kawaguchi T, Berger MS, Pieper RO: DNA damaging agent-induced autophagy produces a cytoprotective adenosine triphosphate surge in malignant glioma cells. Cell Death Differ. 2007, 14 (3): 548-558. 10.1038/sj.cdd.4402030.CrossRefPubMed Katayama M, Kawaguchi T, Berger MS, Pieper RO: DNA damaging agent-induced autophagy produces a cytoprotective adenosine triphosphate surge in malignant glioma cells. Cell Death Differ. 2007, 14 (3): 548-558. 10.1038/sj.cdd.4402030.CrossRefPubMed
6.
go back to reference Baehrecke EH: Autophagy: dual roles in life and death?. Nat Rev Mol Cell Biol. 2005, 6 (6): 505-510. 10.1038/nrm1666.CrossRefPubMed Baehrecke EH: Autophagy: dual roles in life and death?. Nat Rev Mol Cell Biol. 2005, 6 (6): 505-510. 10.1038/nrm1666.CrossRefPubMed
7.
go back to reference Lum JJ, DeBerardinis RJ, Thompson CB: Autophagy in metazoans: cell survival in the land of plenty. Nat Rev Mol Cell Biol. 2005, 6 (6): 439-448. 10.1038/nrm1660.CrossRefPubMed Lum JJ, DeBerardinis RJ, Thompson CB: Autophagy in metazoans: cell survival in the land of plenty. Nat Rev Mol Cell Biol. 2005, 6 (6): 439-448. 10.1038/nrm1660.CrossRefPubMed
8.
go back to reference Yorimitsu T, Klionsky DJ: Eating the endoplasmic reticulum: quality control by autophagy. Trends Cell Biol. 2007, 17 (6): 279-285. 10.1016/j.tcb.2007.04.005.CrossRefPubMed Yorimitsu T, Klionsky DJ: Eating the endoplasmic reticulum: quality control by autophagy. Trends Cell Biol. 2007, 17 (6): 279-285. 10.1016/j.tcb.2007.04.005.CrossRefPubMed
9.
go back to reference Aita VM, Liang XH, Murty VV, Pincus DL, Yu W, Cayanis E, Kalachikov S, Gilliam TC, Levine B: Cloning and genomic organization of beclin1, a candidate tumor suppressor gene on chromosome 17q21. Genomics. 1999, 59 (1): 59-65. 10.1006/geno.1999.5851.CrossRefPubMed Aita VM, Liang XH, Murty VV, Pincus DL, Yu W, Cayanis E, Kalachikov S, Gilliam TC, Levine B: Cloning and genomic organization of beclin1, a candidate tumor suppressor gene on chromosome 17q21. Genomics. 1999, 59 (1): 59-65. 10.1006/geno.1999.5851.CrossRefPubMed
10.
go back to reference Miracco C, Cosci E, Oliveri G, Luzi P, Pacenti L, Monciatti I, Mannucci S, De Nisi MC, Toscano M, Malagnino V, Falzarano SM, Pirtoli L, Tosi P: Protein and mRNA expression of autophagy gene Beclin 1 in human brain tumours. Int J Onco. 2007, 30 (2): 429-436. Miracco C, Cosci E, Oliveri G, Luzi P, Pacenti L, Monciatti I, Mannucci S, De Nisi MC, Toscano M, Malagnino V, Falzarano SM, Pirtoli L, Tosi P: Protein and mRNA expression of autophagy gene Beclin 1 in human brain tumours. Int J Onco. 2007, 30 (2): 429-436.
11.
go back to reference Mizushima N, Noda T, Yoshimori T, Tanaka Y, Ishii T, George MD, Klionsky DJ, Ohsumi M, Ohsumi Y: A protein conjugation system essential for autophagy. Nature. 1998, 395 (6700): 395-398. 10.1038/26506.CrossRefPubMed Mizushima N, Noda T, Yoshimori T, Tanaka Y, Ishii T, George MD, Klionsky DJ, Ohsumi M, Ohsumi Y: A protein conjugation system essential for autophagy. Nature. 1998, 395 (6700): 395-398. 10.1038/26506.CrossRefPubMed
12.
go back to reference Ravikumar B, Imarisio S, Sarkar S, O’Kane CJ, Rubinsztein DC: Rab5 Modulates aggregation and toxicity of mutant huntingtin through macroautophagy in cell and fly models of Huntington disease. Cell Sci. 2008, 121 (Pt 10): 1649-1660.CrossRef Ravikumar B, Imarisio S, Sarkar S, O’Kane CJ, Rubinsztein DC: Rab5 Modulates aggregation and toxicity of mutant huntingtin through macroautophagy in cell and fly models of Huntington disease. Cell Sci. 2008, 121 (Pt 10): 1649-1660.CrossRef
13.
go back to reference Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H, Levine B: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 1999, 402 (6762): 672-676. 10.1038/45257.CrossRefPubMed Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H, Levine B: Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 1999, 402 (6762): 672-676. 10.1038/45257.CrossRefPubMed
14.
go back to reference Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T: LC3, A mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000, 19 (21): 5720-5728. 10.1093/emboj/19.21.5720.PubMedCentralCrossRefPubMed Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T: LC3, A mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000, 19 (21): 5720-5728. 10.1093/emboj/19.21.5720.PubMedCentralCrossRefPubMed
15.
go back to reference Luo S, Rubinsztein DC: Apoptosis blocks beclin 1-dependent autophagosome synthesis: an effect rescued by Bcl-xL. Cell Death Differ. 2010, 17 (2): 268-277. 10.1038/cdd.2009.121.PubMedCentralCrossRefPubMed Luo S, Rubinsztein DC: Apoptosis blocks beclin 1-dependent autophagosome synthesis: an effect rescued by Bcl-xL. Cell Death Differ. 2010, 17 (2): 268-277. 10.1038/cdd.2009.121.PubMedCentralCrossRefPubMed
16.
go back to reference Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A, Rosen J, Eskelinen EL, Mizushima N, Ohsumi Y, Cattoretti G, Levine B: Promotion of tumorigenesis by heterozygous disruption of the beclin1 autophagy gene. Clin Invest. 2003, 112 (12): 1809-1820.CrossRef Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A, Rosen J, Eskelinen EL, Mizushima N, Ohsumi Y, Cattoretti G, Levine B: Promotion of tumorigenesis by heterozygous disruption of the beclin1 autophagy gene. Clin Invest. 2003, 112 (12): 1809-1820.CrossRef
17.
go back to reference Boya P, González-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N, Métivier D, Meley D, Souquere S, Yoshimori T, Pierron G, Codogno P, Kroemer G: Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol. 2005, 25 (3): 1025-1040. 10.1128/MCB.25.3.1025-1040.2005.PubMedCentralCrossRefPubMed Boya P, González-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N, Métivier D, Meley D, Souquere S, Yoshimori T, Pierron G, Codogno P, Kroemer G: Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol. 2005, 25 (3): 1025-1040. 10.1128/MCB.25.3.1025-1040.2005.PubMedCentralCrossRefPubMed
18.
go back to reference Wang ZH, Peng ZL, Duan ZL, Liu H: Expression and clinical significance of autophagy gene Beclin1 in cervical squamous cell carcinoma. Sichuan Da Xue Xue Bao Yi Xue Ban. 2006, 37 (6): 860-863.PubMed Wang ZH, Peng ZL, Duan ZL, Liu H: Expression and clinical significance of autophagy gene Beclin1 in cervical squamous cell carcinoma. Sichuan Da Xue Xue Bao Yi Xue Ban. 2006, 37 (6): 860-863.PubMed
19.
go back to reference Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD, Levine B: Bcl-2 antiapoptotic proteins inhibit beclin 1-dependent autophagy. Cell. 2005, 122 (6): 927-939. 10.1016/j.cell.2005.07.002.CrossRefPubMed Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD, Levine B: Bcl-2 antiapoptotic proteins inhibit beclin 1-dependent autophagy. Cell. 2005, 122 (6): 927-939. 10.1016/j.cell.2005.07.002.CrossRefPubMed
20.
go back to reference Saleem A, Dvorzhinski D, Santanam U, Mathew R, Bray K, Stein M, White E, DiPaola RS: Effect of dual inhibition of apoptosis and autophagy in prostate cancer. Prostate. 2012, 72 (12): 1374-1381. 10.1002/pros.22487.CrossRefPubMed Saleem A, Dvorzhinski D, Santanam U, Mathew R, Bray K, Stein M, White E, DiPaola RS: Effect of dual inhibition of apoptosis and autophagy in prostate cancer. Prostate. 2012, 72 (12): 1374-1381. 10.1002/pros.22487.CrossRefPubMed
Metadata
Title
Beclin1 inhibition promotes autophagy and decreases gemcitabine–induced apoptosis in Miapaca2 pancreatic cancer cells
Authors
Xiaoshu Li
Jun Yan
Lisheng Wang
Fengjun Xiao
Yuefeng Yang
Xiaozhong Guo
Hua Wang
Publication date
01-12-2013
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2013
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/1475-2867-13-26

Other articles of this Issue 1/2013

Cancer Cell International 1/2013 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine