Determination of Apoptotic Stage of HCC827 Lung Adenocarcinoma Cell Line, Following Calcium Sulfide Nanoclusters Treatment

Abstract

Lung cancer is the number one cause of death independently of gender. By the time of the initial diagnosis, more than 50% of all lung cancer patients have already developed secondary tumors due to metastases. Despite current treatments such as surgery, chemotherapy, and radiotherapy, the prognosis for these patients still poor. Subsequently, there exists the need for more efficient treatments in order to increase the survival rate of patients suffering this disease. Previously, in‐vitro experiments in our lab demonstrated that Calcium Sulfide (CaS) nanoclusters induced apoptosis and necrosis in the non‐small lung cell adenocarcinoma (NSCLC) cell line HCC827. Using the Flow Cytometry technology, in this project we aimed to determine the effect of CaS in these cells evaluating apoptotic markers including, Caspase‐3/7 activity, Bcl‐2 phosphorylation, mitochondria potential, DNA‐damage, and oxidative stress. We hypothesized that CaS will show significant differences in these assays when compared to the Dimethyl sulfoxide (DMSO) control. We added a single dose of CaS [3.8μM] (experimental treatment), DMSO (control vehicle) or Etoposide [10 nM] (positive control) at time 0 and then collected the data after 24, 48, and 72 hours. As our results, we did not observe any statistically significant effect on apoptotic markers, Caspase‐3/7 and Bcl‐2. Analysis of the Mitopotential assay showed an statistical increase (p≤0.01) of dead cells population at 48 hours and of depolarized/lives cells at 72 hours compared to DMSO, in HCC827 cells treated with CaS. However, at 48 hours cells treated with CaS decrease the numbers of non‐affected cells (p≤0.05), increase DNA double‐strand breaks (p≤0.001), decrease the activation of Ataxia‐telangiectasia mutated kinase (ATM) and the phosphorylation of H2A.X significantly (p≤0.01). These results suggested that the detrimental effect of CaS on malignant HCC827 cell line is influenced by the accumulation of oxidative stress damage also affecting the cell cycle as our lab previously reported. Additional studies are ongoing to validate these results. Within the next month we plan to study the lung non‐malignant cells MRC‐5 using the same assays.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.