Piperlongumine potentiates the antitumor efficacy of oxaliplatin through ROS induction in gastric cancer cells
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Oxaliplatin is one of the most commonly used chemotherapeutic agents in the treatment of various cancers, including gastric cancer. It has, however, a narrow therapeutic index due to its toxicity and the occurrence of drug resistance. Therefore, there is a pressing need to develop novel therapies to potentiate the efficacy and reduce the toxicity of oxaliplatin. Piperlongumine (PL), an alkaloid isolated from Piper longum L., has recently been identified as a potent agent against cancer cells in vitro and in vivo. In the present study, we investigated whether PL can potentiate the antitumor effect of oxaliplatin in gastric cancer cells.
Cellular apoptosis and ROS levels were analyzed by flow cytometry. Thioredoxin reductase 1 (TrxR1) activity in gastric cancer cells or tumor tissues was determined using an endpoint insulin reduction assay. Western blotting was used to analyze the expression levels of the indicated proteins. Nude mice xenograft models were used to test the effects of PL and oxaliplatin combinations on gastric cancer cell growth in vivo.
We found that PL significantly enhanced oxaliplatin-induced growth inhibition in both gastric and colon cancer cells. Moreover, we found that PL potentiated the antitumor effect of oxaliplatin by inhibiting TrxR1 activity. PL combined with oxaliplatin markedly suppressed the activity of TrxR1, resulting in the accumulation of ROS and, thereby, DNA damage induction and p38 and JNK signaling pathway activation. Pretreatment with antioxidant N-acetyl-L-cysteine (NAC) significantly abrogated the combined treatment-induced ROS generation, DNA damage and apoptosis. Importantly, we found that activation of the p38 and JNK signaling pathways prompted by PL and oxaliplatin was also reversed by NAC pretreatment. In vivo, we found that PL combined with oxaliplatin significantly suppressed tumor growth in a gastric cancer xenograft model, and effectively reduced the activity of TrxR1 in tumor tissues. Remarkably, we found that PL attenuated body weight loss evoked by oxaliplatin treatment.
Our data support a synergistic effect of PL and oxaliplatin and suggest that application of its combination may be more effective for the treatment of gastric cancer than oxaliplatin alone.
KeywordsGastric cancer Thioredoxin reductase 1 Reactive oxygen species Piperlongumine Oxaliplatin
Reactive oxygen species
Thioredoxin reductase 1
c-Jun N-terminal kinase
p38 mitogen-activated protein kinase.
Peng Zou and Yiqun Xia designed the study, supervised the project and wrote the manuscript. Peichen Zhang, Lingyan Shi and Tingting Zhang designed and performed most of the experiments. Lin Hong carried out the immunofluorescence experiments. Wei He, Peihai Cao, Xin Shen and Peisen Zheng collected and analyzed the data. All authors were involved in the final version of the manuscript.
This research was supported by the National Natural Science Foundation of China (81603153 and 81503107) and the Zhejiang Provincial Natural Science Foundation of China (LY16H310011).
Compliance with ethical standards
The authors declare that they have no competing interests.
Ethics approval and consent to participate
The experimental protocol was established according to the Guide for the Care and Use of Laboratory Animals, and was approved by the Institutional Animal Care and Use Committee of Wenzhou Medical University.
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