Nitric Oxide Donor DETA/NO Inhibits the Growth of Endometrial Cancer Cells by Upregulating the Expression of RASSF1 and CDKN1A.

Sana Waheed,David A Wink,Viqar Syed,Yovanni Casablanca,Robert Ys Cheng,G Larry Maxwell

doi:10.3390/molecules24203722

Abstract

Nitric oxide (NO) is implicated in several biological processes, including cancer progression. At low concentrations, it promotes cell survival and tumor progression, and at high concentrations it causes apoptosis and cell death. Until now, the impact of NO donors has not been investigated on human endometrial tumors. Four cancer cell lines were exposed to different concentrations of DETA/NO for 24 to 120 h. The effects of DETA/NO on cell proliferation and invasion were determined utilizing MTS and Boyden chamber assays, respectively. The DETA/NO induced a dose and time-dependent reduction in cell viability by the activation of caspase-3 and cell cycle arrest at the G0/G1 phase that was associated with the attenuated expression of cyclin-D1 and D3. Furthermore, the reduction in the amount of CD133-expressing cancer stem-like cell subpopulation was observed following DETA/NO treatment of cells, which was associated with a decreased expression of stem cell markers and attenuation of cell invasiveness. To understand the mechanisms by which DETA/NO elicits anti-cancer effects, RNA sequencing (RNA-seq) was used to ascertain alterations in the transcriptomes of human endometrial cancer cells. RNA-seq analysis revealed that 14 of the top 21 differentially expressed genes were upregulated and seven were downregulated in endometrial cancer cells with DETA/NO. The genes that were upregulated in all four cell lines with DETA/NO were the tumor suppressors Ras association domain family 1 isoform A (RASSF1) and Cyclin-dependent kinase inhibitor 1A (CDKN1A). The expression patterns of these genes were confirmed by Western blotting. Taken together, the results provide the first evidence in support of the anti-cancer effects of DETA/NO in endometrial cancer.

Highlights

Endometrial carcinoma ranks as the fourth most common cancer in women in America, with61,880 new cases and 12,160 deaths anticipated in 2019 [1]
To investigate the inhibitory effects of DETA/Nitric oxide (NO), four endometrial cancer cells were treated with various doses of DETA/NO for 24, 48, 72, 96, and 120 h, and cell viability was assessed using MTS
Some cell lines were more responsive to DETA/NO than others

Summary

Introduction

Endometrial carcinoma ranks as the fourth most common cancer in women in America, with61,880 new cases and 12,160 deaths anticipated in 2019 [1]. There is an increase in the number of patients in an advanced stage or with a high histological grade, which reflects poor prognosis. The treatment of early-stage disease, commonly consisting of surgery followed by chemotherapy, is very effective with a cure rate of 90%. Over two-thirds of patients are diagnosed with advanced stages, in which. Nitric oxide (NO), a free radical, is involved in several diseases including cancer [2]. It is endogenously synthesized in various tissues by the transformation of L-arginine to L-citrulline in three distinct isoforms of nitric oxide synthase (NOS). Inducible nitric oxide synthase (iNOS) is the calcium-independent isoform and is induced by cytokines, endotoxins, and hypoxia under oxidative stress. Inducible nitric oxide synthase (iNOS) is the calcium-independent isoform and is induced by cytokines, endotoxins, and hypoxia under oxidative stress. iNOS produces significant concentrations (micromolar range) of NO for a longer period of time ranging from hours to days [4,5]

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