Cisplatin Induces Apoptosis Through the Endoplasmic Reticulum-mediated, Calpain 1 Pathway in Triple-negative Breast Cancer Cells

Shadia M. Al-Bahlani, Khadija H. Al-Bulushi, Zaina M. Al-Alawi, Nadia Y. Al-Abri, Zuweina R. Al-Hadidi, Shaikha S. Al-Rawahi

Research output: Contribution to journalArticle

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Abstract

Chemoresistance of triple-negative breast cancer (TNBC) is a major obstacle for successful treatment and is mainly represented as a defect in apoptosis. Using advanced techniques in molecular biology, we show that calpain 1 plays an essential role in modulating TNBC cell sensitivity to cisplatin-induced apoptosis. Exploring new pathways in cisplatin-induced apoptosis will help in overcoming the resistance to apoptosis in TNBC cells. Background Breast cancer is the most common cancer in women worldwide. Triple-negative breast cancer (TNBC) is an aggressive type that can be treated using platinum-based chemotherapy such as cisplatin (cis-diamminedichloroplatinum II). Although the calpain protein is essential in many cellular processes, including apoptosis, cell signaling, and proliferation, its role in cisplatin-induced apoptosis in TNBC cells is not fully understood. The present study assessed calpain 1–dependent, cisplatin-induced apoptosis in TNBC cells. Materials and Methods MDA-MB231 cells were treated with different concentrations of cisplatin (0, 20, and 40 μM). The cisplatin deposit and its effect on endoplasmic reticulum and, subsequently, calcium release were detected using transmission electron microscopy and Von Koss staining, respectively. Calpain 1 messenger RNA, protein content, and apoptosis was measured using reverse transcriptase-polymerase chain reaction, Western blotting, and Hoechst stain, respectively. In addition, calpain modulation, by either activation or inhibition, and its effect on cisplatin-induced apoptosis were assessed. Results Our results showed that cisplatin induced endoplasmic reticulum stress, indicated by an increase in calcium staining and protein expression of glucose-regulated protein 78 and calmodulin, followed by cleavage of α-fodrin and caspase-12 and, eventually, apoptosis. Cyclopiazonic acid showed a similar effect and enhanced the sensitivity of these cells to cisplatin treatment. In contrast, calpain 1 inhibition by both specific small interfering RNA and exogenous inhibitor (calpeptin) attenuated cisplatin-induced apoptosis in these cells. Conclusion Altogether, these findings suggest, for the first time, that calpain 1 activation by endoplasmic reticulum plays an essential role in sensitizing TNBC cells to cisplatin-induced apoptosis. This finding will allow exploration of new insights for the treatment of TNBC by overcoming its resistance to apoptosis.

Original languageEnglish
Pages (from-to)e103-e112
JournalClinical Breast Cancer
Volume17
Issue number3
DOIs
Publication statusPublished - Jun 1 2017

Fingerprint

Triple Negative Breast Neoplasms
Calpain
Endoplasmic Reticulum
Cisplatin
Apoptosis
Caspase 12
Staining and Labeling
Calcium
Proteins
Endoplasmic Reticulum Stress
Calmodulin
Platinum
Reverse Transcriptase Polymerase Chain Reaction
Transmission Electron Microscopy
Small Interfering RNA

Keywords

  • Breast cancer
  • Calcium-dependent proteases
  • Electron microscope
  • Programmed cell death
  • Western blot

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Cisplatin Induces Apoptosis Through the Endoplasmic Reticulum-mediated, Calpain 1 Pathway in Triple-negative Breast Cancer Cells. / Al-Bahlani, Shadia M.; Al-Bulushi, Khadija H.; Al-Alawi, Zaina M.; Al-Abri, Nadia Y.; Al-Hadidi, Zuweina R.; Al-Rawahi, Shaikha S.

In: Clinical Breast Cancer, Vol. 17, No. 3, 01.06.2017, p. e103-e112.

Research output: Contribution to journalArticle

Al-Bahlani, Shadia M. ; Al-Bulushi, Khadija H. ; Al-Alawi, Zaina M. ; Al-Abri, Nadia Y. ; Al-Hadidi, Zuweina R. ; Al-Rawahi, Shaikha S. / Cisplatin Induces Apoptosis Through the Endoplasmic Reticulum-mediated, Calpain 1 Pathway in Triple-negative Breast Cancer Cells. In: Clinical Breast Cancer. 2017 ; Vol. 17, No. 3. pp. e103-e112.
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abstract = "Chemoresistance of triple-negative breast cancer (TNBC) is a major obstacle for successful treatment and is mainly represented as a defect in apoptosis. Using advanced techniques in molecular biology, we show that calpain 1 plays an essential role in modulating TNBC cell sensitivity to cisplatin-induced apoptosis. Exploring new pathways in cisplatin-induced apoptosis will help in overcoming the resistance to apoptosis in TNBC cells. Background Breast cancer is the most common cancer in women worldwide. Triple-negative breast cancer (TNBC) is an aggressive type that can be treated using platinum-based chemotherapy such as cisplatin (cis-diamminedichloroplatinum II). Although the calpain protein is essential in many cellular processes, including apoptosis, cell signaling, and proliferation, its role in cisplatin-induced apoptosis in TNBC cells is not fully understood. The present study assessed calpain 1–dependent, cisplatin-induced apoptosis in TNBC cells. Materials and Methods MDA-MB231 cells were treated with different concentrations of cisplatin (0, 20, and 40 μM). The cisplatin deposit and its effect on endoplasmic reticulum and, subsequently, calcium release were detected using transmission electron microscopy and Von Koss staining, respectively. Calpain 1 messenger RNA, protein content, and apoptosis was measured using reverse transcriptase-polymerase chain reaction, Western blotting, and Hoechst stain, respectively. In addition, calpain modulation, by either activation or inhibition, and its effect on cisplatin-induced apoptosis were assessed. Results Our results showed that cisplatin induced endoplasmic reticulum stress, indicated by an increase in calcium staining and protein expression of glucose-regulated protein 78 and calmodulin, followed by cleavage of α-fodrin and caspase-12 and, eventually, apoptosis. Cyclopiazonic acid showed a similar effect and enhanced the sensitivity of these cells to cisplatin treatment. In contrast, calpain 1 inhibition by both specific small interfering RNA and exogenous inhibitor (calpeptin) attenuated cisplatin-induced apoptosis in these cells. Conclusion Altogether, these findings suggest, for the first time, that calpain 1 activation by endoplasmic reticulum plays an essential role in sensitizing TNBC cells to cisplatin-induced apoptosis. This finding will allow exploration of new insights for the treatment of TNBC by overcoming its resistance to apoptosis.",
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N2 - Chemoresistance of triple-negative breast cancer (TNBC) is a major obstacle for successful treatment and is mainly represented as a defect in apoptosis. Using advanced techniques in molecular biology, we show that calpain 1 plays an essential role in modulating TNBC cell sensitivity to cisplatin-induced apoptosis. Exploring new pathways in cisplatin-induced apoptosis will help in overcoming the resistance to apoptosis in TNBC cells. Background Breast cancer is the most common cancer in women worldwide. Triple-negative breast cancer (TNBC) is an aggressive type that can be treated using platinum-based chemotherapy such as cisplatin (cis-diamminedichloroplatinum II). Although the calpain protein is essential in many cellular processes, including apoptosis, cell signaling, and proliferation, its role in cisplatin-induced apoptosis in TNBC cells is not fully understood. The present study assessed calpain 1–dependent, cisplatin-induced apoptosis in TNBC cells. Materials and Methods MDA-MB231 cells were treated with different concentrations of cisplatin (0, 20, and 40 μM). The cisplatin deposit and its effect on endoplasmic reticulum and, subsequently, calcium release were detected using transmission electron microscopy and Von Koss staining, respectively. Calpain 1 messenger RNA, protein content, and apoptosis was measured using reverse transcriptase-polymerase chain reaction, Western blotting, and Hoechst stain, respectively. In addition, calpain modulation, by either activation or inhibition, and its effect on cisplatin-induced apoptosis were assessed. Results Our results showed that cisplatin induced endoplasmic reticulum stress, indicated by an increase in calcium staining and protein expression of glucose-regulated protein 78 and calmodulin, followed by cleavage of α-fodrin and caspase-12 and, eventually, apoptosis. Cyclopiazonic acid showed a similar effect and enhanced the sensitivity of these cells to cisplatin treatment. In contrast, calpain 1 inhibition by both specific small interfering RNA and exogenous inhibitor (calpeptin) attenuated cisplatin-induced apoptosis in these cells. Conclusion Altogether, these findings suggest, for the first time, that calpain 1 activation by endoplasmic reticulum plays an essential role in sensitizing TNBC cells to cisplatin-induced apoptosis. This finding will allow exploration of new insights for the treatment of TNBC by overcoming its resistance to apoptosis.

AB - Chemoresistance of triple-negative breast cancer (TNBC) is a major obstacle for successful treatment and is mainly represented as a defect in apoptosis. Using advanced techniques in molecular biology, we show that calpain 1 plays an essential role in modulating TNBC cell sensitivity to cisplatin-induced apoptosis. Exploring new pathways in cisplatin-induced apoptosis will help in overcoming the resistance to apoptosis in TNBC cells. Background Breast cancer is the most common cancer in women worldwide. Triple-negative breast cancer (TNBC) is an aggressive type that can be treated using platinum-based chemotherapy such as cisplatin (cis-diamminedichloroplatinum II). Although the calpain protein is essential in many cellular processes, including apoptosis, cell signaling, and proliferation, its role in cisplatin-induced apoptosis in TNBC cells is not fully understood. The present study assessed calpain 1–dependent, cisplatin-induced apoptosis in TNBC cells. Materials and Methods MDA-MB231 cells were treated with different concentrations of cisplatin (0, 20, and 40 μM). The cisplatin deposit and its effect on endoplasmic reticulum and, subsequently, calcium release were detected using transmission electron microscopy and Von Koss staining, respectively. Calpain 1 messenger RNA, protein content, and apoptosis was measured using reverse transcriptase-polymerase chain reaction, Western blotting, and Hoechst stain, respectively. In addition, calpain modulation, by either activation or inhibition, and its effect on cisplatin-induced apoptosis were assessed. Results Our results showed that cisplatin induced endoplasmic reticulum stress, indicated by an increase in calcium staining and protein expression of glucose-regulated protein 78 and calmodulin, followed by cleavage of α-fodrin and caspase-12 and, eventually, apoptosis. Cyclopiazonic acid showed a similar effect and enhanced the sensitivity of these cells to cisplatin treatment. In contrast, calpain 1 inhibition by both specific small interfering RNA and exogenous inhibitor (calpeptin) attenuated cisplatin-induced apoptosis in these cells. Conclusion Altogether, these findings suggest, for the first time, that calpain 1 activation by endoplasmic reticulum plays an essential role in sensitizing TNBC cells to cisplatin-induced apoptosis. This finding will allow exploration of new insights for the treatment of TNBC by overcoming its resistance to apoptosis.

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