Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress

Kosuke Sakitani, Yoshihiro Hirata, Yohko Hikiba, Yoku Hayakawa, Sozaburo Ihara, Hirobumi Suzuki, Nobumi Suzuki, Takako Serizawa, Hiroto Kinoshita, Kei Sakamoto, Hayato Nakagawa, Keisuke Tateishi, Shin Maeda, Tsuneo Ikenoue, Shoji Kawazu, Kazuhiko Koike

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background: Although some molecularly targeted drugs for colorectal cancer are used clinically and contribute to a better prognosis, the current median survival of advanced colorectal cancer patients is not sufficient. Autophagy, a basic cell survival mechanism mediated by recycling of cellular amino acids, plays an important role in cancer. Recently, autophagy has been highlighted as a promising new molecular target. The unfolded protein response (UPR) reportedly act in complementary fashion with autophagy in intestinal homeostasis. However, the roles of UPR in colon cancer under autophagic inhibition remain to be elucidated. We aim to clarify the inhibitory effect of autophagy on colon cancer. Methods: We crossed K19 CreERT and Atg5 flox/flox mice to generate Atg5 flox/flox /K19 CreERT mice. Atg5 flox/flox /K19 CreERT mice were first treated with azoxymethane/dextran sodium sulfate and then injected with tamoxifen to inhibit autophagy in CK19-positive epithelial cells. To examine the anti-cancer mechanisms of autophagic inhibition, we used colon cancer cell lines harboring different p53 gene statuses, as well as small interfering RNAs (siRNAs) targeting Atg5 and immunoglobulin heavy-chain binding protein (BiP), a chaperone to aid folding of unfolded proteins. Results: Colon tumors in Atg5 flox/flox /K19 CreERT mice showed loss of autophagic activity and decreased tumor size (the total tumor diameter was 28.1 mm in the control and 20.7 mm in Atg5 flox/flox /K19 CreERT mice, p = 0.036). We found that p53 and UPR/endoplasmic reticulum (ER) stress-related proteins, such as cleaved caspase 3, and CAAT/enhancer-binding protein homologous protein, are up-regulated in colon tumors of Atg5 flox/flox /K19 CreERT mice. Although Atg5 and BiP silencing, respectively, increased apoptosis in p53 wild type cells, Atg5 silencing alone did not show the same effect on apoptosis in p53 mutant cells. However, co-transfection of Atg5 and BiP siRNAs led to increased apoptosis in p53 mutant cells. Conclusions: Blocking autophagy has potential in the treatment of colon cancer by inducing apoptosis via p53 and ER stress, and suppressing the UPR pathway is a valid strategy to overcome resistance to autophagic inhibition.

LanguageEnglish
Article number795
JournalBMC Cancer
Volume15
Issue number1
DOIs
Publication statusPublished - 24 Oct 2015
Externally publishedYes

Keywords

  • Apoptosis
  • Autophagy
  • Colon cancer

ASJC Scopus subject areas

  • Oncology
  • Genetics
  • Cancer Research

Cite this

Sakitani, K., Hirata, Y., Hikiba, Y., Hayakawa, Y., Ihara, S., Suzuki, H., ... Koike, K. (2015). Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress. BMC Cancer, 15(1), [795]. https://doi.org/10.1186/s12885-015-1789-5
Sakitani, Kosuke ; Hirata, Yoshihiro ; Hikiba, Yohko ; Hayakawa, Yoku ; Ihara, Sozaburo ; Suzuki, Hirobumi ; Suzuki, Nobumi ; Serizawa, Takako ; Kinoshita, Hiroto ; Sakamoto, Kei ; Nakagawa, Hayato ; Tateishi, Keisuke ; Maeda, Shin ; Ikenoue, Tsuneo ; Kawazu, Shoji ; Koike, Kazuhiko. / Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress. In: BMC Cancer. 2015 ; Vol. 15, No. 1.
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title = "Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress",
abstract = "Background: Although some molecularly targeted drugs for colorectal cancer are used clinically and contribute to a better prognosis, the current median survival of advanced colorectal cancer patients is not sufficient. Autophagy, a basic cell survival mechanism mediated by recycling of cellular amino acids, plays an important role in cancer. Recently, autophagy has been highlighted as a promising new molecular target. The unfolded protein response (UPR) reportedly act in complementary fashion with autophagy in intestinal homeostasis. However, the roles of UPR in colon cancer under autophagic inhibition remain to be elucidated. We aim to clarify the inhibitory effect of autophagy on colon cancer. Methods: We crossed K19 CreERT and Atg5 flox/flox mice to generate Atg5 flox/flox /K19 CreERT mice. Atg5 flox/flox /K19 CreERT mice were first treated with azoxymethane/dextran sodium sulfate and then injected with tamoxifen to inhibit autophagy in CK19-positive epithelial cells. To examine the anti-cancer mechanisms of autophagic inhibition, we used colon cancer cell lines harboring different p53 gene statuses, as well as small interfering RNAs (siRNAs) targeting Atg5 and immunoglobulin heavy-chain binding protein (BiP), a chaperone to aid folding of unfolded proteins. Results: Colon tumors in Atg5 flox/flox /K19 CreERT mice showed loss of autophagic activity and decreased tumor size (the total tumor diameter was 28.1 mm in the control and 20.7 mm in Atg5 flox/flox /K19 CreERT mice, p = 0.036). We found that p53 and UPR/endoplasmic reticulum (ER) stress-related proteins, such as cleaved caspase 3, and CAAT/enhancer-binding protein homologous protein, are up-regulated in colon tumors of Atg5 flox/flox /K19 CreERT mice. Although Atg5 and BiP silencing, respectively, increased apoptosis in p53 wild type cells, Atg5 silencing alone did not show the same effect on apoptosis in p53 mutant cells. However, co-transfection of Atg5 and BiP siRNAs led to increased apoptosis in p53 mutant cells. Conclusions: Blocking autophagy has potential in the treatment of colon cancer by inducing apoptosis via p53 and ER stress, and suppressing the UPR pathway is a valid strategy to overcome resistance to autophagic inhibition.",
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author = "Kosuke Sakitani and Yoshihiro Hirata and Yohko Hikiba and Yoku Hayakawa and Sozaburo Ihara and Hirobumi Suzuki and Nobumi Suzuki and Takako Serizawa and Hiroto Kinoshita and Kei Sakamoto and Hayato Nakagawa and Keisuke Tateishi and Shin Maeda and Tsuneo Ikenoue and Shoji Kawazu and Kazuhiko Koike",
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Sakitani, K, Hirata, Y, Hikiba, Y, Hayakawa, Y, Ihara, S, Suzuki, H, Suzuki, N, Serizawa, T, Kinoshita, H, Sakamoto, K, Nakagawa, H, Tateishi, K, Maeda, S, Ikenoue, T, Kawazu, S & Koike, K 2015, 'Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress', BMC Cancer, vol. 15, no. 1, 795. https://doi.org/10.1186/s12885-015-1789-5

Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress. / Sakitani, Kosuke; Hirata, Yoshihiro; Hikiba, Yohko; Hayakawa, Yoku; Ihara, Sozaburo; Suzuki, Hirobumi; Suzuki, Nobumi; Serizawa, Takako; Kinoshita, Hiroto; Sakamoto, Kei; Nakagawa, Hayato; Tateishi, Keisuke; Maeda, Shin; Ikenoue, Tsuneo; Kawazu, Shoji; Koike, Kazuhiko.

In: BMC Cancer, Vol. 15, No. 1, 795, 24.10.2015.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Inhibition of autophagy exerts anti-colon cancer effects via apoptosis induced by p53 activation and ER stress

AU - Sakitani, Kosuke

AU - Hirata, Yoshihiro

AU - Hikiba, Yohko

AU - Hayakawa, Yoku

AU - Ihara, Sozaburo

AU - Suzuki, Hirobumi

AU - Suzuki, Nobumi

AU - Serizawa, Takako

AU - Kinoshita, Hiroto

AU - Sakamoto, Kei

AU - Nakagawa, Hayato

AU - Tateishi, Keisuke

AU - Maeda, Shin

AU - Ikenoue, Tsuneo

AU - Kawazu, Shoji

AU - Koike, Kazuhiko

PY - 2015/10/24

Y1 - 2015/10/24

N2 - Background: Although some molecularly targeted drugs for colorectal cancer are used clinically and contribute to a better prognosis, the current median survival of advanced colorectal cancer patients is not sufficient. Autophagy, a basic cell survival mechanism mediated by recycling of cellular amino acids, plays an important role in cancer. Recently, autophagy has been highlighted as a promising new molecular target. The unfolded protein response (UPR) reportedly act in complementary fashion with autophagy in intestinal homeostasis. However, the roles of UPR in colon cancer under autophagic inhibition remain to be elucidated. We aim to clarify the inhibitory effect of autophagy on colon cancer. Methods: We crossed K19 CreERT and Atg5 flox/flox mice to generate Atg5 flox/flox /K19 CreERT mice. Atg5 flox/flox /K19 CreERT mice were first treated with azoxymethane/dextran sodium sulfate and then injected with tamoxifen to inhibit autophagy in CK19-positive epithelial cells. To examine the anti-cancer mechanisms of autophagic inhibition, we used colon cancer cell lines harboring different p53 gene statuses, as well as small interfering RNAs (siRNAs) targeting Atg5 and immunoglobulin heavy-chain binding protein (BiP), a chaperone to aid folding of unfolded proteins. Results: Colon tumors in Atg5 flox/flox /K19 CreERT mice showed loss of autophagic activity and decreased tumor size (the total tumor diameter was 28.1 mm in the control and 20.7 mm in Atg5 flox/flox /K19 CreERT mice, p = 0.036). We found that p53 and UPR/endoplasmic reticulum (ER) stress-related proteins, such as cleaved caspase 3, and CAAT/enhancer-binding protein homologous protein, are up-regulated in colon tumors of Atg5 flox/flox /K19 CreERT mice. Although Atg5 and BiP silencing, respectively, increased apoptosis in p53 wild type cells, Atg5 silencing alone did not show the same effect on apoptosis in p53 mutant cells. However, co-transfection of Atg5 and BiP siRNAs led to increased apoptosis in p53 mutant cells. Conclusions: Blocking autophagy has potential in the treatment of colon cancer by inducing apoptosis via p53 and ER stress, and suppressing the UPR pathway is a valid strategy to overcome resistance to autophagic inhibition.

AB - Background: Although some molecularly targeted drugs for colorectal cancer are used clinically and contribute to a better prognosis, the current median survival of advanced colorectal cancer patients is not sufficient. Autophagy, a basic cell survival mechanism mediated by recycling of cellular amino acids, plays an important role in cancer. Recently, autophagy has been highlighted as a promising new molecular target. The unfolded protein response (UPR) reportedly act in complementary fashion with autophagy in intestinal homeostasis. However, the roles of UPR in colon cancer under autophagic inhibition remain to be elucidated. We aim to clarify the inhibitory effect of autophagy on colon cancer. Methods: We crossed K19 CreERT and Atg5 flox/flox mice to generate Atg5 flox/flox /K19 CreERT mice. Atg5 flox/flox /K19 CreERT mice were first treated with azoxymethane/dextran sodium sulfate and then injected with tamoxifen to inhibit autophagy in CK19-positive epithelial cells. To examine the anti-cancer mechanisms of autophagic inhibition, we used colon cancer cell lines harboring different p53 gene statuses, as well as small interfering RNAs (siRNAs) targeting Atg5 and immunoglobulin heavy-chain binding protein (BiP), a chaperone to aid folding of unfolded proteins. Results: Colon tumors in Atg5 flox/flox /K19 CreERT mice showed loss of autophagic activity and decreased tumor size (the total tumor diameter was 28.1 mm in the control and 20.7 mm in Atg5 flox/flox /K19 CreERT mice, p = 0.036). We found that p53 and UPR/endoplasmic reticulum (ER) stress-related proteins, such as cleaved caspase 3, and CAAT/enhancer-binding protein homologous protein, are up-regulated in colon tumors of Atg5 flox/flox /K19 CreERT mice. Although Atg5 and BiP silencing, respectively, increased apoptosis in p53 wild type cells, Atg5 silencing alone did not show the same effect on apoptosis in p53 mutant cells. However, co-transfection of Atg5 and BiP siRNAs led to increased apoptosis in p53 mutant cells. Conclusions: Blocking autophagy has potential in the treatment of colon cancer by inducing apoptosis via p53 and ER stress, and suppressing the UPR pathway is a valid strategy to overcome resistance to autophagic inhibition.

KW - Apoptosis

KW - Autophagy

KW - Colon cancer

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U2 - 10.1186/s12885-015-1789-5

DO - 10.1186/s12885-015-1789-5

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JO - BMC cancer

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