Suppression of autophagy enhances preferential toxicity of paclitaxel to folliculin-deficient renal cancer cells

Journal of Experimental & Clinical Cancer Research

Guan Wu³  Xun-Bo Jin²  Jindong Chen¹  Sue Schoen¹  Shuhui Si¹  Qi Zhang¹ 

[1] Department of Urology, University of Rochester Medical Center, 601 Elmwood Avenue, Box 656, 14642 Rochester, NY, USA;Minimally Invasive Urology Center, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China;Wilmot Cancer Center, University of Rochester Medical Center, Rochester, NY, USA

关键词: Kidney cancer;    BHD;    FLCN;    Folliculin;    Taxol;    Paclitaxel;    Apoptosis;    Autophagy;   

Others  :  814066
DOI  :  10.1186/1756-9966-32-99

 received in 2013-09-23, accepted in 2013-11-21,  发布年份 2013

【 摘 要 】

Background

Paclitaxel, a widely used chemotherapeutic drug, can induce apoptosis in variety of cancer cells. A previous study has shown preferential toxicity of paclitaxel to FLCN-deficient kidney cancer cell line, UOK257. In this report, we investigate the cellular and molecular mechanism of paclitaxel-induced autophagy and apoptosis in renal cancer cells with and without FLCN expression.

Methods

Two pairs of cell lines were used: FLCN siRNA-silenced ACHN cell line (ACHN-5968) and scrambled ACHN cell line (ACHN-sc); FLCN-null UOK257 cell line and UOK257-2 cell line restored with ectopic expression of FLCN. Autophagy was examined by western blot, GFP-LC3, transmission electron microscopy, and MDC assay. Cell viability and apoptosis were detected using MTT assay, DAPI stain and TUNEL assay. After inhibition of autophagy with 3-Methyladenine (3-MA) or Beclin 1 siRNA, cell viability and apoptosis were measured by MTT assay and TUNEL assay.

Results

After paclitaxel treatment, a dose-dependent decrease in cell viability and increase in apoptosis were observed in FLCN-deficient UOK257 and ACHN-5968 cells compared to their FLCN-expressing counterparts, suggesting that renal cancer cells without FLCN were more sensitive to paclitaxel. Enhanced autophagy was found to be associated with paclitaxel treatment in FLCN-deficient RCC cells. The MAPK pathway was also identified as a key pathway for the activation of autophagy in these kidney cancer cells. Inhibition of phosphorylated ERK with ERK inhibitor U0126 showed a significant decrease in autophagy. Furthermore, after inhibition of autophagy with 3-Methyladenine (3-MA) or Beclin 1 siRNA, apoptosis induced by paclitaxel was significantly increased in FLCN-deficient UOK257 and ACHN-5968 cells.

Conclusions

Preferential toxicity of paclitaxel to FLCN-deficient kidney cancer cells is associated with enhanced autophagy. Suppression of autophagy further enhances paclitaxel-induced apoptosis in FLCN-deficient renal cancer cells. Our results suggest that paclitaxel combined with an autophagy inhibitor might be a potentially more effective chemotherapeutic approach for FLCN-deficient renal cancer.

【 授权许可】

   
2013 Zhang et al.; licensee BioMed Central Ltd.

【 图 表 】

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