In vitro toxicity assay of cisplatin on mouse acute lymphoblastic leukaemia and spermatogonial stem cells

Andrologia. 2016 Jun;48(5):584-94. doi: 10.1111/and.12490. Epub 2015 Oct 1.

Abstract

Testicular cancer is the most common cancer affecting men in reproductive age, and cisplatin is one of the major helpful chemotherapeutic agents for treatment of this cancer. In addition, exposure of testes cancer cells to cisplatin could potentially eliminate tumour cells from germ cells in patients. The aim of this study was to evaluate the effect of cisplatin on viability of mouse acute lymphoblastic leukaemia cell line (EL-4) and neonatal mouse spermatogonial cells in vitro. In this study, the isolated spermatogonial stem cells (SSC) and EL-4 were divided into six groups including control (received medium), sham (received DMSO in medium) and experimental groups which received different doses of cisplatin (0.5, 5, 10 and 15 μg ml(-1) ). Cells viability was evaluated with MTT assay. The identity of the cultured cells was confirmed by the expression of specific markers. Our finding showed that viability of both SSC and EL-4 cells was reduced with the dose of 15 μg/ml when compared to the control group (P ≤ 0.05). Also, the differences between the IC50 in doses 10 and 15 μg/ml at different time were significant (P ≤ 0.05). The number of TUNEL-positive cells was increased, and the BAX and caspase-3 expressions were upregulated in EL4 cells for group that received an effective dose of cisplatin). In conclusion, despite the dramatic effects of cisplatin on both cells, spermatogonial stem cells could form colony in culture.

Keywords: Cisplatin; EL4 cells; SSCs; TUNEL assay; viability.

MeSH terms

  • Adult Germline Stem Cells / cytology
  • Adult Germline Stem Cells / drug effects*
  • Adult Germline Stem Cells / metabolism
  • Animals
  • Animals, Newborn
  • Antineoplastic Agents / administration & dosage
  • Antineoplastic Agents / toxicity*
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Caspase 3 / genetics
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cisplatin / administration & dosage
  • Cisplatin / toxicity*
  • Colony-Forming Units Assay
  • Gene Expression / drug effects
  • Humans
  • In Vitro Techniques
  • Male
  • Mice
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / drug therapy*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / genetics
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / pathology
  • Spermatogenesis / drug effects
  • bcl-2-Associated X Protein / genetics

Substances

  • Antineoplastic Agents
  • Bax protein, mouse
  • bcl-2-Associated X Protein
  • Casp3 protein, mouse
  • Caspase 3
  • Cisplatin