Induction of Apoptosis in the Human Prostate Cancer Cell Line DU-145 by a Novel Micronutrient Formulation

M Waheed Roomi,Matthias Rath,Neha Shanker,Aleksandra Niedzwiecki

doi:10.4236/ojapo.2015.41002

Abstract

Prostate cancer, the most frequently diagnosed cancer in men, primarily affects males aged 55 and older and is more common in African Americans than Caucasians. Once the cancer has metastasized, current treatments are generally ineffective. We have identified a novel anti-neoplastic agent, a specifically designed nutrient mixture (NM), containing ascorbic acid, lysine, proline and green tea extract that demonstrates a broad spectrum of anti-tumor activity against a number of human cancer cell lines. In a previous study NM significantly inhibited prostate tumor in nude mice. In this study, we tested whether the formulation exerts its anti-tumor effects through induction of apoptosis on prostate cancer cell line DU-145. The effect of the nutrient mixture (NM) on cell growth inhibition in DU-145 cells was examined by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Morphological changes and caspase activation associated with apoptosis induction was checked by H&E staining and Live Green Caspase assay, respectively. The NM was found to be slightly toxic to DU-145 cells at 100 μg/ml, but significantly toxic at 500 μg/ml and 1000 μg/ml. Percentage of cells undergoing apoptosis also increased from 6% at 100 μg/ml to 49% at 500 μg/ml and 83% at 1000 μg/ml, with greater number of cells showing morphological changes such as condensed nuclei and an acidophilic cytoplasm at higher concentrations. For the purpose of comparison, NM was also tested on a normal human dermal fibroblast (NHDF) cell line which exhibited far less apoptosis induction as compared to DU-145 cells. The percentage of cells undergoing apoptosis in case of NHDF cells was 7% at 100 μg/ml, 25.6% at 500 μg/ml and 76.5% at 1000 μg/ml. Our results demonstrate that the NM is effective in inhibiting cancer cell viability and inducing apoptosis in prostate cancer DU-145 cells and can thus be used as an effective treatment for prostate cancer.

Highlights

Prostate cancer, the leading non-cutaneous cancer and the second leading cause of death in men in the U.S, primarily affects males of age 55 and above and is more common in African American males than white males [1]
In this study we investigated in a DU-145 prostate cancer cell line whether the inhibition of prostate tumors by nutrient mixture (NM) was due to induction of apoptosis
Nutrient Mixture Decreased the Viability of Prostate Cancer Cells To determine whether the nutrient formulation is cytotoxic to prostate cancer cells DU-145, we used the MTT assay to check for cell viability at various NM dosages

Summary

Introduction

The leading non-cutaneous cancer and the second leading cause of death in men in the U.S, primarily affects males of age 55 and above and is more common in African American males than white males [1]. It accounted for approximately 15% of all cancer diagnosis and 6.6% of all cancer deaths in men in 2012. Standard treatment of prostate cancer consists of surgery (prostatectomy), hormonal therapy and radiotherapy. While prostate carcinomas are initially responsive to surgery and hormonal therapies when localized, they eventually become more aggressive and unresponsive to standard treatments

Methods

Results

Conclusion