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Promoting Effect of Adriamycin, Cisplatinum and Etoposide on Trk Genes Expression in Human Neuroblastoma Cells

Author(s): Ling Liu, Yang Li, Xilin Xiong, Chi Zhang, Jianpei Fang

Purpose: Tropomyosin receptor kinases (Trk) gene family had been strongly correlated with tumor progression and found to be highly predictive of clinical behavior. The aim of the study is to evaluated the cytotoxicity and cell cycle effect of adriamycin (ADM), cisplatinum (DDP) and etoposide (VP16) on neuroblastoma(NB) cells, and then assessed effect of three anti-cancer agents on Trk gene family expression in NB in vitro conditions.

Methods: SK-N-SH NB cells were exposed in vitro to ADM, DDP and VP16. Cell counting kit (CCK-8) assay was performed to determine the effect of anti-cancer agents on cell viability, and the cell cycle change was measured by flow cytometry. Quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis was utilized to detect the expression levels of Trk gene family (TrkA, TrkB and TrkC) in a wide range of concentration of ADM, DDP and VP16 at different time.

Results: Our data illustrated that ADM, DDP and VP16 had cytotoxic activity as a single agent in both a time- and dose-dependent manner in vitro. However, NB cells resistance to high concentrations of ADM. It was confirmed that NB cells showed in vitro sensitivity to ADM, DDP and Vp16, with concentration that observed with the half-maximal inhibitory concentration (IC50) values on SK-N-SH cells being 5μg/ml, 8μg/ml and 100μg/ml, respectively. Flow cytometry analysis showed that these three drugs also leaded to enhanced accumulation of cell populations in S phase. Additionally, our results showed that in NB cells, these three drugs treatment dramatically increased expression of TrkA, TrkB and TrkC in RT-PCR analysis.

Conclusions: Collectively, our results may help to develop tailored treatment approaches and enhance the efficacy of cytotoxic agents treatment for high-risk NB with minimal or no additional toxicity by adjusting the concentration and action time of ADM, DDP and VP16 depending on different NB stages.

    Editor In Chief

    Masashi Emoto

  • Professor of Laboratory of Immunology
    Department of Laboratory Sciences
    Gunma University Graduate School of Health Sciences
    Gunma, Japan

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