# Effects of Spiro-bisheterocycles on Proliferation and Apoptosis in Human Breast Cancer Cell Lines

Lamia Hamdan Ramdani, Oualid Tahli, Laetitia Delort, Caroline Decombat, Arthur Silva, Khaldoun Bachari, Marie-Paule Vasson, Florence Caldefie-Chezet

► To cite this version:

Lamia Hamdan Ramdani, Oualid Tahli, Laetitia Delort, Caroline Decombat, Arthur Silva, et al. Effects of Spiro-bisheterocycles on Proliferation and Apoptosis in Human Breast Cancer Cell Lines. Fourth International Congress of Translational Research in Human Nutrition ICTRHN 2017, Jun 2017, Clermont-Ferrand, France. 45 p., 2017, 4th International Congress of Translational Research in Human Nutrition & Cancer:Abstract Book. hal-01594048

HAL Id: hal-01594048

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Submitted on 26 Sep 2017

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# Effects of Spiro-*bisheterocycles* on Proliferation and Apoptosis in Human Breast Cancer Cell Lines

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UNIVERSITE
Clermont
Auvergne

Lamia HAMDAN RAMDANI<fnref n="1" />,<fnref n="2" />, Qualid TALHI<fnref n="1" />,<fnref n="4" />, Nadia TAIBI<fnref n="1" />, Laetitia DELORT<fnref n="2" />, Caroline DECOMBAT<fnref n="2" />, Artur SILVA<fnref n="4" />,
Khaldoun BACHARI<fnref n="1" />, Marie-Paule VASSON<fnref n="2" />,<fnref n="3" />, and Florence CALDEFIE-CHEZET<fnref n="2" />

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## Context and Objective

Breast cancer is the first cause of cancer death in women worldwide and a major concern to public health. Targeting p53-MDM2 interaction by using appropriate molecules offers an attractive strategy for p53 activation and a promising approach for new anticancer therapies. In this context, we provide an account of the evaluation of small molecules containing oxygen and nitrogen spiro-*bisheterocycles* in terms of effects on *in vitro* proliferation and apoptosis of human breast cancer cell lines (MCF-7 and MDA-MB231). The wider aim is to identify novel low-molecular-weight, easily-accessible synthetic spiro compounds for cancer chemotherapy.

## Experimental Procedures

* Spiro-*bisheterocyclic* compounds 2-7 were chemically synthesized.

* Cell proliferation was assessed by resazurin assay.

* Annexin V-FITC/ PI apoptosis assays was performed in order to demonstrate apoptotic effects of spiro-*bisheterocyclic* in breast cancer cell lines.

* Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to analyze the wild-type p53, MDM2, BAX and caspase 3 gene expression levels.

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## Results

<figure><img src="image_6.png" /><figcaption>Figure 1. Effect of compounds 2-7 on the *in vitro* growth and proliferation of human breast cancer cells. MCF-7 and MDA-MB231 cell line were treated with 10, 25, 50 and 100 µM of compounds 2-7 for 72 h.</figcaption></figure>

The treatment with compounds 3 and 6, corresponding to spiro [hydantoin-isoxazole] and spiro [hydantoin-oxazepine] respectively, resulted in a significant decrease of cell proliferation and the induction of the apoptosis in both breast cancer cell lines whereas spiro [hydantoin-diazepine] was only active against MDA-MB 231 (Figure 1).

<figure><img src="image_7.png" /><figcaption>Figure 2. Drug-induced apoptosis in breast cancer cell lines MCF-7 and MDA-MB-231. Bright-field (top row), and fluorescent images of annexin V-fluorescein isothiocyanate (FITC) (middle row) and propidium iodide (PI) (bottom row) staining.</figcaption></figure>

The images show significant induction of apoptosis with compounds 3 and 6 in MCF-7 cells, while strong apoptosis was observed with compounds 3, 4 and 6 in MDA-MB-231 cells (Figure 2).

<figure><img src="image_8.png" /><figcaption>Figure 3; Results of qRT-PCR detection of p53, MDM2, BAX and caspase-3 gene expression in MCF-7 and MDA-MB-231 cells, following treatment with 100 µM of compounds 3, 4 and 6 for 72 h.</figcaption></figure>

The qRT-PCR revealed an up-regulation of MDM2, strictly p53-dependent, and an increase of the expression of pro-apoptotic genes such as caspase 3 and BAX in MCF-7 wild-type p53 and MDA-MB-231 mutant p53 breast cancer cells (Figure 3).

## Conclusion

In summary, this study brings the first demonstration that new small spiro-*bisheterocyclic* molecules can sensitize breast cancer cells to apoptosis by targeting p53-MDM2 interaction. We also highlighted that these spiro compounds promote apoptosis via p53-independent pathway(s), suggesting that these compounds represent interesting candidates as therapeutic targets for the treatment of breast cancer.

<footnote n="1">¹CENTRE DE RECHERCHE SCIENTIFIQUE ET TECHNIQUE EN ANALYSES PHYSICO-CHIMIQUES (CRAPC), TIPAZA, ALGÉRIE;</footnote>

<footnote n="2">²UNIVERSITÉ CLERMONT AUVERGNE, INRA, UNH, UNITÉ DE NUTRITION HUMAINE, CRNH AUVERGNE, F-63000 CLERMONT-FERRAND, FRANCE</footnote>

<footnote n="3">³CENTRE JEAN-PERRIN, CLERMONT-FERRAND, FRANCE;</footnote>

<footnote n="4">⁴ORGANIC CHEMISTRY, NATURAL PRODUCTS AND AGRIFOOD (QOPNA), DEPARTMENT OF CHEMISTRY & QOPNA, UNIVERSITY OF AVEIRO, AVEIRO, PORTUGAL</footnote>