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Bortezomib-induced apoptosis in cultured pancreatic cancer cells is associated with ceramide production

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Abstract

Purpose

The proteasome inhibitor bortezomib (PS-341) has displayed significant efficiency against pancreatic cancer cells. However, the underlying mechanisms are not fully understood. Here, we tested if ceramide production was involved in the bortezomib’s effect.

Methods

Two transformed pancreatic cancer cell lines (PANC-1 and Mia) and the primary pancreatic cancer cells were used. Cell death was analyzed by MTT viability assay and trypan blue staining. Cell apoptosis was analyzed by Histone DNA-ELISA assay and Annexin V FACS. Western blots were used to test signal protein changes. The cellular ceramide level after bortezomib treatment was also determined.

Results

In cultured pancreatic cancer cells, bortezomib increased cellular ceramide production to promote cell apoptosis. The ceramide de novo synthase inhibitor fumonisin B1 (F-B1) suppressed bortezomib-induced ceramide production and apoptosis, while exogenously added C6-ceramide facilitated bortezomib-induced pancreatic cancer cell death. Meanwhile, 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), the inhibitor of glucosylceramide synthetase as well as the sphingosine kinase 1 inhibitors (SKI-II and SKI-IV), facilitated bortezomib-induced ceramide production and subsequent cell apoptosis. Further, bortezomib-induced pro-apoptotic c-Jun N-terminal kinase (JNK) activation was also associated with ceramide production. JNK activation by bortezomib was suppressed by F-B1, but was enhanced by SKI-II and PDMP in pancreatic cancer cells. Finally, C6-ceramide, SKI-II, and PDMP dramatically enhanced bortezomib-induced cytotoxicity in primary cultured pancreatic cancer cells.

Conclusions

We found that bortezomib-induced apoptosis was associated with ceramide production in primary and transformed pancreatic cancer cells.

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Abbreviations

JNK:

C-Jun N-terminal kinase

SphK1:

Sphingosine kinase 1

F-B1:

Fumonisin B1

PDMP:

1-Phenyl-2-decanoylamino-3-morpholino-1-propanol

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Authors and Affiliations

  1. Department of Gastroenterology, Wuxi No. 2 People’s Hospital, Wuxi, Jiangsu, China

    Lei Gong, Xuejun Tang, Yan Jing & Gao-jue Wu

  2. Department of Gastroenterology, East Hospital Affiliated to Tongji University, No. 150, Ji-mo Road, Pudong New District, Shanghai, 200120, China

    Ming Xu, Bo Cheng & Ping Zheng

  3. Department of Surgery, The Third Hospital Affiliated to Soochow University, Changzhou City, 213003, Jiangsu, China

    Bo Yang

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  1. Lei Gong
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Correspondence to Ming Xu.

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Lei Gong, Bo Yang and Ming Xu have contributed equally to this work.

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Gong, L., Yang, B., Xu, M. et al. Bortezomib-induced apoptosis in cultured pancreatic cancer cells is associated with ceramide production. Cancer Chemother Pharmacol 73, 69–77 (2014). https://doi.org/10.1007/s00280-013-2318-3

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  • DOI: https://doi.org/10.1007/s00280-013-2318-3

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