BCL2A1 regulates Canady Helios Cold Plasma-induced cell death in triple-negative breast cancer

Saravana R K Murthy,Michael Keidar,Lawan Ly,Olivia Jones,Xiaoqian Cheng,Jerome Canady,Taisen Zhuang,Giacomo Basadonna

doi:10.1038/s41598-022-07027-4

Abstract

Breast cancer is the leading cause of cancer death among women. Triple-negative breast cancer (TNBC) has a poor prognosis and frequently relapses early compared with other subtypes. The Cold Atmospheric Plasma (CAP) is a promising therapy for prognostically poor breast cancer such as TNBC. The Canady Helios Cold Plasma (CHCP) induces cell death in the TNBC cell line without thermal damage, however, the mechanism of cell death by CAP treatment is ambiguous and the mechanism of resistance to cell death in some subset of cells has not been addressed. We investigate the expression profile of 48 apoptotic and 35 oxidative gene markers after CHCP treatment in six different types of breast cancer cell lines including luminal A (ER+ PR+/−HER2−), luminal B (ER+PR+/−HER2+), (ER−PR−HER2+), basal-like: ER−PR−HER2− cells were tested with CHCP at different power settings and at 4 different incubation time. The expression levels of the gene markers were determined at 4 different intervals after the treatment. The protein expression of BCL2A1 was only induced after CHCP treatment in TNBC cell lines (p < 0.01), whereas the HER2-positive and ER, PR positive cell lines showed little or no expression of BCL2A1. The BCL2A1 and TNF-alpha expression levels showed a significant correlation within TNBC cell lines (p < 0.01). Silencing BCL2A1 mRNA by siRNA increased the potency of the CHCP treatment. A Combination of CHCP and CPI203, a BET bromodomain inhibitor, and a BCL2A1 antagonist increased the CHCP-induced cell death (p < 0.05). Our results revealed that BCL2A1 is a key gene for resistance during CHCP induced cell death. This resistance in TNBCs could be reversed with a combination of siRNA or BCL2A1 antagonist-CHCP therapy.

Highlights

Breast cancer is the most common cause of cancer death among women w orldwide[1] and it exhibits diverse molecular features that reflect the high heterogeneity which complicates the clinical treatment[2]
To systematically investigate the molecular basis for survival after Cold Atmospheric Plasma (CAP) treatment by breast cancer cell lines which are classified into their intrinsic subtypes such as luminal A (ER+progesterone receptor (PR)+/−human epidermal growth factor receptor 2 (HER2)−), luminal B (ER+PR+/-HER2+), basal-like (ER−PR−HER2−), and HER2-positive (ER−PR−HER2+), we conducted quantitative real time PCR analysis to screen the genes that are differentially expressed after Canady Helios Cold Plasma (CHCP) treatment
ABL proto-oncogene 1, non-receptor tyrosine kinase (ABL1), Fas-associated via death domain (FADD) and Tumor Protein p53 (TP53) mRNA expression were significantly (p < 0.05) down-regulated after CHCP treatment compared to untreated mock samples, whereas death associated protein kinase 1 (DAPK1) expression was not detected in either of the sample groups

Summary

Introduction

Breast cancer is the most common cause of cancer death among women w orldwide[1] and it exhibits diverse molecular features that reflect the high heterogeneity which complicates the clinical treatment[2]. At high concentrations and duration of CHCP treatment, most of the breast cancer cells often undergo apoptosis due to the release of reactive oxygen and nitrogen species (RONS) and oxidative stress-induced cell toxicity of these s pecies[15]. The mechanism of such oxidative stress-induced cell death process is broadly discussed in various studies[16–18]. Our previous studies of CHCP treatment on subtypes of breast cancer cell lines showed that its ability to reduce breast cancer viability by 92–99% regardless of the status of the receptors on these cells at the most optimal power setting and time of t reatment[12]. This study was undertaken to identify genes that could predict response to CHCP treatment on subtypes of breast cancer cell lines and the genes that instigate resistance to cell death induced by CHCP treatment

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Results

Conclusion