Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Annals of Surgical Oncology
Published in: Annals of Surgical Oncology 3/2009

01-03-2009 | Laboratory and Translational Research

Inhibition of Autophagy by 3-MA Enhances the Effect of 5-FU-Induced Apoptosis in Colon Cancer Cells

Authors: Jie Li, MD, Ni Hou, MD, PhD, Ahmad Faried, MD, PhD, Soichi Tsutsumi, MD, PhD, Toshiyuki Takeuchi, MD, PhD, Hiroyuki Kuwano, MD, PhD

Published in: Annals of Surgical Oncology | Issue 3/2009

Login to get access

Abstract

Background

5-fluorouracil-(5-FU)-based adjuvant chemotherapy is widely used for the treatment of colorectal cancer. However, 5-FU resistance in the course of treatment has become more common. Therefore, new therapeutic strategies and/or new adjuvant drugs still need to be explored.

Methods

Two colon-cancer-derived cell lines, colon26 and HT29, were used to investigate the effect of 5-FU, 3-methyladenine (3-MA, an autophagy inhibitor), or their combination on apoptotic cell death and autophagy. MTT assay, Hochest plus propidium iodide (PI) staining, and DNA fragmentation assay were used to observe apoptosis. Meanwhile, monodansylcadaverine (MDC) was used to detect autophagy. Finally, immunoblotting assay was used to explore the molecular change that occurred.

Results

We observed the apoptosis induced by 5-FU in colon cancer cells. Meanwhile, autophagy was also stimulated. The combination treatment of 3-MA and 5-FU significantly increased the apoptotic cell death. By isolating the subcellular fractions of mitochondria and cytosol, we observed that the release of cytochrome c was increased in combination-treated cells. Cytochrome c resulted in the activation of caspase-3, thus activating PARP. Moreover, the anti-apoptotic protein, Bcl-xL, was significantly downregulated by 3-MA.

Conclusions

Our results suggest that 5-FU-induced apoptosis in colon cancer cells can be enhanced by the inhibitor of autophagy, 3-MA. Autophagy might play a role as a self-defense mechanism in 5-FU-treated colon cancer cells, and its inhibition could be a promising strategy for the adjuvant chemotherapy of colon cancer.
Literature
1.
Xu R, Zhou B, Fung PCW, Li X. Recent advances in the treatment of colon cancer. Histol Histopathol. 2006;21:867–72.PubMed
2.
Chau I, Cunningham D. Adjuvant therapy in colon cancer: current status and future directions. Cancer Treat Rev. 2002;28:223–36.PubMedCrossRef
3.
Chau I, Cunningham D. Adjuvant therapy in colon cancer-what, when and how? Ann Oncol. 2006;17:1347–59.PubMedCrossRef
4.
5.
Nordman IC, Iyer S, Joshua AM, Clarke SJ. Advance in the adjuvant treatment of colorectal cancer. ANZ J Surg. 2006;76:373–80.PubMedCrossRef
6.
Wils J, O’Dwyer P, Labianca R. Adjuvant treatment of colorectal cancer at the turn of the century: European and US perspectives. Ann Oncol. 2001;12:13–22.PubMedCrossRef
7.
Giacchetti BS, Perpoint B, Zidani R, et al. Phase III multicenter randomized trial of oxaliplatin added to chronomodulated fluorouracil–leucovorin as first-line treatment of metastatic colorectal cancer. J Clin Oncol. 2000;18:136–47.PubMed
8.
Douillard JY, Cunningham D, Roth AD, et al. Irinotecan combined with fluorouracil compared with fluorouracil alone as first-line treatment for metastatic colorectal cancer: a multicentre randomised trial. Lancet. 2000;355:1041–7.PubMedCrossRef
9.
Longley DB, Harkin DP, Johnston PG. 5-Fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer. 2003;3:330–8.PubMedCrossRef
10.
Grem JL. 5-fluorouracil: forty-plus and still ticking. A review of its preclinical and clinical development. Invest New Drugs. 2000;18:299–313.PubMedCrossRef
11.
12.
Petiot A, Ogier-Denis E, Blommaart EFC, Meijer AJ, Codogno P. Distinct classes of phosphatidylinositol 3′ kinases are involved in signaling pathways that control macroautophagy in HT–29 cells. J Biol Chem. 2000;275:992–8.PubMedCrossRef
13.
Yan CH, Liang ZQ, Gu ZL, Yang YP, Reid P, Qin ZH. Contributions of autophagic and apoptotic mechanisms to CrTX induced death of K562 cells. Toxicon. 2006;47:521–30.PubMedCrossRef
14.
Cui Q, Tashiro S, Onodera S, Ikejima T. Augmentation of oridonin-induced apoptosis observed with reduced autophagy. J Pharmacol Sci. 2006;101:230–9.PubMedCrossRef
15.
Klionsky DJ, Emr SD. Autophagy as a regulated pathway of cellular degradation. Science. 2000;290:1717–21.PubMedCrossRef
16.
Meijer AJ, Codogno P. Regulation and role of autophagy in mammalian cells. Int J Biochem Cell B. 2004;36:2445–62.CrossRef
17.
Ng G, Huang JX. The significance of autophagy in cancer. Mol Carcinogen. 2005;43:183–7.CrossRef
18.
Ogier-Denis E, Codogno P. Autophagy: a barrier or an adaptive response to cancer. BBA-Rev Cancer. 2003;1603:113–28.
19.
Kondo Y, Kanzawa T, Sawaya R, Kondo S. The role of autophagy in cancer development and response to therapy. Nat Rev Cancer. 2005;5:726–34.PubMedCrossRef
20.
Hoorens A, Casteele MV, Klöppel G, Pipeleers D. Glucose promotes survival of rat pancreatic cells by activating synthesis of proteins which suppress a constitutive apoptotic program. J Clin Invest. 1996;98:1568–74.PubMedCrossRef
21.
Munafó DB, Colombo MI. A novel assay to study autophagy: regulation of autophagosome vacuole size by amino acid deprivation. J Cell Sci. 2001;114:3619–29.PubMed
22.
Abedin MJ, Wang D, McDonnell MA, Lehmann U, Kelekar A. Autophagy delays apoptotic cell death in breast cancer cells following DNA damage. Cell Death Differ. 2006;22: 1–11.
23.
Biederbick A, Kern HF, Elsasser HP. Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles. Eur J Cell Biol. 1995;66:3–14.PubMed
24.
Shimizu S, Kanaseki T, Mizushima N, Mizuta T, Arakawa-Kobayashi S, Thompson CB, et al. Role of Bcl–2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes. Nat Cell Biol. 2004;6:1221–8.PubMedCrossRef
25.
Herman-Antosiewicz A, Johnson DE, Singh SV. Sulforaphane causes autophagy to inhibit release of cytochrome c and apoptosis in human prostate cancer cells. Cancer Res. 2006;66:5828–35.PubMedCrossRef
26.
Klionsky DJ, Levine B. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev Cell. 2004;6:463–77.PubMedCrossRef
27.
Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H, et al. Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature. 1999;402:672–6.PubMedCrossRef
28.
Qu XP, Yu J, Bhagat G, et al. Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. J Clin Invest. 2003;112:1809–20.PubMed
29.
Ding WX, Ni HM, Gao WT, et al. Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival. J Biol Chem. 2007;282:4702–10.PubMedCrossRef
30.
Bauvy C, Gane P, Arico S, Codogon P, Ogier-Denis E. Autophagy delays Sulindac Sulfide-induced apoptosis in the human intestinal colon cancer cell line HT-29. Exp Cell Res. 2001;268:139–49.PubMedCrossRef
31.
Kanzawa T, Germano IM, Komata T, Kondo Y, Kondo S. Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ. 2004;11:448–57.PubMedCrossRef
32.
Paglin S, Hollister T, Delohery T, et al. A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res. 2001;61:439–44.PubMed
33.
Kanzawa T, Kondo Y, Ito H, Kondo S, Germano I. Induction of autophagic cell death in malignant glioma cells by arsenic trioxide. Cancer Res. 2003;63:2103–8.PubMed
34.
35.
Bursch W, Ellinger A, Kienzl H, et al. Active cell death induced by the anti-estrogens tamoxifen and ICI164 384 in human mammary carcinoma cells (MCF–7) in culture: the role of autophagy. Carcinogenesis. 1996;17:1595–607.PubMedCrossRef
36.
Cao C, Subhawong T, Albert JM, et al. Inhibition of mammalian target of rapamycin or apoptotic pathway induces autophagy and radiosensitizes PTEN null prostate cancer cells. Cancer Res. 2006;66:10040–7.PubMedCrossRef
37.
Iwamaru A, Kondo Y, Iwado E, et al. Silencing mammalian target of rapamycin signaling by small interfering RNA enhances rapamycin-induced autophagy in malignant glioma cells. Oncogene. 2007;26:1840–51.PubMedCrossRef
38.
Zhu HB, Guo W, Zhang LD, et al. Bcl-xL small interfering RNA suppresses the proliferation of 5-fluorouracil-resistant human colon cancer cells. Mol Cancer Ther. 2005;4:451–6.PubMedCrossRef
39.
Tsujimoto Y. Cell death regulation by the Bcl–2 protein family in the mitochondria. J Cell Physiol. 2003;195:158–67.PubMedCrossRef
40.
Pattingre S, Tassa A, Qu XP, et al. Bcl-2 antiapoptotic proteins inhibit beclin 1-dependent autophagy. Cell. 2005;122:927–39.PubMedCrossRef
Metadata
Title
Inhibition of Autophagy by 3-MA Enhances the Effect of 5-FU-Induced Apoptosis in Colon Cancer Cells
Authors
Jie Li, MD
Ni Hou, MD, PhD
Ahmad Faried, MD, PhD
Soichi Tsutsumi, MD, PhD
Toshiyuki Takeuchi, MD, PhD
Hiroyuki Kuwano, MD, PhD
Publication date
01-03-2009
Publisher
Springer-Verlag
Published in
Annals of Surgical Oncology / Issue 3/2009
Print ISSN: 1068-9265
Electronic ISSN: 1534-4681
DOI
https://doi.org/10.1245/s10434-008-0260-0

Other articles of this Issue 3/2009

Annals of Surgical Oncology 3/2009 Go to the issue

Melanomas

Surveillance Chest X-Rays for Melanoma: Prescription or Proscription?

Gastrointestinal Oncology

In Reply: Circulating Cancer Cells and Their Clinical Use as Recurrence Markers for Gastric and Colorectal Cancer

Breast Oncology

Personal Genomics and Genome-Wide Association Studies: Novel Discoveries but Limitations for Practical Personalized Medicine

Breast Oncology

Preoperative Chemotherapy Decreases the Need for Re-Excision of Breast Cancers Between 2 and 4 cm Diameter

Laboratory and Translational Research

Clinical Implications of the Mechanisms Driving Breast Cancer Local Recurrence

Gastrointestinal Oncology

The Prognostic Impact of Isolated Tumor Cells in Lymph Nodes of T2N0 Gastric Cancer: Comparison of American and Japanese Gastric Cancer Patients