Increased intracellular Ca²⁺ decreases cisplatin resistance by regulating iNOS expression in human ovarian cancer cells - 17/01/17

Doi : 10.1016/j.biopha.2016.11.135

Yang Yu ^a,^{^{1
These authors contributed equally to this manuscript.}}, Qi Xie ^b,^{^{1
These authors contributed equally to this manuscript.}}, Weimin Liu ^c, Yuting Guo ^a, Na Xu ^a, Lu Xu ^a, Shibing Liu ^a, Songyan Li ^a, Ye Xu ^a,^⁎ , Liankun Sun ^b,^⁎
^a Key Laboratory of Colleges and Universities of Jilin Province, Basic College of Medicine, Jilin Medical University, Jilin, Jilin, China
^b Department of Pathophysiology, Basic College of Medicine, Jilin University, Changchun, Jilin, China
^c Department of Intervention, The Jilin municipal People's Hospital, Jilin, Jilin, China

^⁎Corresponding author at: Key Laboratory of Colleges and Universities of Jilin Province, Basic College of Medicine, Jilin Medical University, 5 Jilin Street, Jilin 132013, China.Key Laboratory of Colleges and Universities of Jilin ProvinceBasic College of MedicineJilin Medical UniversityJilinJilinChina^⁎⁎Corresponding author at: Department of Pathophysiology, Basic College of Medicine, Jilin University, 126 Xinmin Street, Changchun 130021, China.Department of PathophysiologyBasic College of MedicineJilin UniversityChangchunJilinChina

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Abstract

Previous studies have reported that intracellular Ca²⁺ signals and inducible nitric oxide synthase (iNOS) are involved in cell apoptosis. However, the role of iNOS in cisplatin resistance in ovarian cancer remains unclear. Here, we demonstrate that SKOV3/DDP ovarian cancer cells were more resistant to cisplatin than were SKOV3 ovarian cancer cells. The expression of intracellular Ca²⁺ and iNOS was more strongly induced by cisplatin in SKOV3 cells than in SKOV3/DDP cells. TAT-conjugated IP3R-derived peptide (TAT-IDP^S) increased cisplatin-induced iNOS expression and apoptosis in SKOV3/DDP cells. 2-Aminoethoxydiphenyl borate (2-APB) decreased cisplatin-induced iNOS expression and apoptosis in SKOV3 cells. Thus, iNOS induction may be a valuable strategy for improving the anti-tumor efficacy of cisplatin in ovarian cancer.

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Keywords : Ovarian cancer, Cisplatin resistance, Intracellular Ca²⁺, iNOS

Plan

Introduction

Materials and methods

Antibodies and drugs

Cell lines

Cellular viability

Immunofluorescence staining and confocal laser microscopy

Cell apoptosis assay

Determination of intracellular Ca²⁺ levels

Western blotting

Statistical analysis

Results

Cisplatin induces higher levels of apoptosis in SKOV3 cells than in SKOV3/DDP cells

The expression of cisplatin-induced intracellular Ca²⁺ and iNOS is higher in SKOV3 cells than in SKOV3/DDP cells

TAT-IDP^S increases cisplatin-induced intracellular Ca²⁺ and iNOS expression in SKOV3/DDP cells

TAT-IDP^S increases cisplatin-induced growth inhibition and apoptosis in SKOV3/DDP cells

2-APB decreases cisplatin-induced intracellular Ca²⁺ and iNOS expression and apoptosis in SKOV3 cells

Discussion

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Increased intracellular Ca²⁺ decreases cisplatin resistance by regulating iNOS expression in human ovarian cancer cells - 17/01/17

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