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Small inhibitor of Bcl-2, HA14-1, selectively enhanced the apoptotic effect of cisplatin by modulating Bcl-2 family members in MDA-MB-231 breast cancer cells

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Abstract

Inhibition or downregulation of Bcl-2 represents a new therapeutic approach to by-pass chemoresistance in cancer cells. Therefore, we explored the potential of this approach in breast cancer cells. Cisplatin and paclitaxel induced apoptosis in a dose-dependent manner in MCF-7 (drug-sensitive) and MDA-MB-231 (drug-insensitive) cells. Furthermore, when we transiently silenced Bcl-2, both cisplatin and paclitaxel induced apoptosis more than parental cells. Dose dependent induction of apoptosis by drugs was enhanced by the pre-treatment of these cells with HA14-1, a Bcl-2 inhibitor. Although the effect of cisplatin was significant on both cell lines, the effect of paclitaxel was much less potent only in MDA-MB-231 cells. To further understand the distinct role of drugs in MDA-MB-231 cells pretreated with HA14-1, caspases and Bcl-2 family proteins were studied. The apoptotic effect of cisplatin with or without HA14-1 pre-treatment is shown to be caspase-dependent. Among pro-apoptotic Bcl-2 proteins, Bax and Puma were found to be up-regulated whereas Bcl-2 and Bcl-xL were down-regulated when cells were pretreated with HA14-1 followed by paclitaxel or cisplatin. Enforced Bcl-2 expression in MDA-MB-231 cells abrogated the sensitizing effect of HA14-1 in cisplatin induced apoptosis. These results suggest that the potentiating effect of HA14-1 is drug and cell type specific and may not only depend on the inhibition of Bcl-2. Importantly, alteration of other pro-apoptotic or anti-apoptotic Bcl-2 family members may dictate the apoptotic response when HA14-1 is combined with chemotherapeutic drugs.

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Acknowledgment

This work was partially supported by Turkish Association for Cancer Research and Control, Terry Fox Cancer Research Grant-35-D/882.

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Authors and Affiliations

  1. Biological Sciences and Bioengineering Program, Faculty of Natural Sciences and Engineering, Sabanci University, Istanbul, Turkey

    Elif Damla Arisan, Ozgur Kutuk, Tugsan Tezil, Cagri Bodur & Huveyda Basaga

  2. Department of Genetics and Bioengineering, Yeditepe University, Kayisdagi, 34755, Istanbul, Turkey

    Dilek Telci

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  1. Elif Damla Arisan
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  2. Ozgur Kutuk
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Correspondence to Huveyda Basaga.

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10549_2009_343_MOESM1_ESM.pdf

Quantitative mRNA levels were determined by quantitative real time PCR assay following transfection of MCF-7 and MDA-MB-231 cells with Bcl-2 siRNA and scramble siRNA for 48 h. Bcl-2 mRNA copy number was calculated using standard curve of β-actin. These values are normalized by dividing with β-actin. Columns represent mean (±SEM) of independent two experiments and each experiment was repeated three times (DOC 22 kb)

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Arisan, E.D., Kutuk, O., Tezil, T. et al. Small inhibitor of Bcl-2, HA14-1, selectively enhanced the apoptotic effect of cisplatin by modulating Bcl-2 family members in MDA-MB-231 breast cancer cells. Breast Cancer Res Treat 119, 271–281 (2010). https://doi.org/10.1007/s10549-009-0343-z

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