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Thioredxin reductase inhibitor ethaselen increases the drug sensitivity of the colon cancer cell line LoVo towards cisplatin via regulation of G1 phase and reversal of G2/M phase arrest

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Summary

We evaluated the combination treatment of ethaselen (BBSKE) as a thioredoxin reductase (TrxR) inhibitor plus cisplatin (CDDP) on the human colon adenocarcinoma cell line LoVo. Therapeutic effects ranging from nearly additive to clearly synergistic demonstrated an effective combination, i.e., the cytostatic dose of CDDP could be reduced without a loss in efficacy. To further investigate the cellular response mechanisms of these favorable outcomes, we analyzed the cell-cycle profiles, mRNA expression patterns, and protein levels of several key genes after incubation with BBSKE or CDDP separately and in combination. In appropriate conditions, CDDP induced arrest at the G2/M phase accompanied by the enhanced inhibitory phosphorylation of Cdk1 and the elevated protein expression of cyclin B1. BBSKE downregulated expression of cyclin D1 by increasing mRNA and protein levels of p21, and thus induced G1 phase arrest. BBSKE returned Cdk1 to an activated state, and reduced the protein level of cyclin B1 after incubation in combination with CDDP, which was consistent with the reduction in the percentage of cells in G2/M identified by flow cytometry. By regulating the G1 phase and reversing CDDP-induced G2/M phase arrest, BBSKE increases drug sensitivity of LoVo cells toward CDDP, and probably provides a meaningful anticancer strategy for further clinical studies.

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Abbreviations

Ethaselen (BBSKE):

1, 2-[bis (1, 2-Benzisoselenazolone-3 (2H)-ketone)] ethane

Cisplatin (CDDP):

cis-diamminedichloroplatinum II

TrxR:

Thioredoxin reductase

Trx:

Thioredoxin

NADPH:

Nicotinamide adenine dinucleotide phosphate

CI:

Combination index

DRI:

Dose-reduction index

GAPDH:

Glyceraldehyde-3-phosphatedehydrogenase

COMB:

Combination

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Acknowledgements

This work was financially supported by the Natural Science Foundation of China (grant number 30472036). We deeply appreciated the help of Professor Chou Ting-Chao (Preclinical Pharmacology Core Laboratory, Memorial Sloan–Kettering Cancer Center, New York, NY, USA) who donated the CompuSyn software and provided careful guidance on the drug combination study.

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Authors and Affiliations

  1. State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, 100191, China

    Jia-ning Fu, Jing Li, Xiao-yuan Ren & Hui-hui Zeng

  2. School of Pharmaceutical Sciences, Peking University, Mailbox 74, Beijing, 100191, China

    Jia-ning Fu, Jing Li, Qiang Tan, Xiao-yuan Ren & Hui-hui Zeng

  3. Peking University First Hospital, Beijing, 100034, China

    Qiang Tan

  4. College of Life Sciences, Peking University, Beijing, 100075, China

    Han-wei Yin, Kun Xiong & Tian-yu Wang

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  1. Jia-ning Fu
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Correspondence to Hui-hui Zeng.

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Jia-ning Fu and Jing Li have contributed equally to this work.

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Fu, Jn., Li, J., Tan, Q. et al. Thioredxin reductase inhibitor ethaselen increases the drug sensitivity of the colon cancer cell line LoVo towards cisplatin via regulation of G1 phase and reversal of G2/M phase arrest. Invest New Drugs 29, 627–636 (2011). https://doi.org/10.1007/s10637-010-9401-y

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