Prostate cancer is the main cause of cancer-related mortality in men around the world and an important health problem. DU-145 human prostate cancer cells provide an opportunity to investigate prostate cancer. Betaine has a number of anticancer effects, such as inactivation of carcinogens, inhibition of cancer cell proliferation, angiogenesis, and metastasis. However, there is no study investigating the effects of betaine on DU-145 cells. The aim of this study was to evaluate the effects of different concentrations of betaine on the oxidative stress, apoptosis, and inflammation on DU-145 cells. Firstly, we proved the cytotoxic activity of betaine (0 to 150 mg/ml) on DU-145 cells by using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and defined the optimal concentration of betaine. Then, by employing the doses found in MTT, the levels of antioxidant (GSH, SOD, CAT, and TAS) and oxidant (MDA and TOS) molecules, pro-inflammatory cytokines (TNF-a and IL-6), apoptotic proteins (CYCS and CASP3), and DNA fragmentation were measured. Morphological changes and apoptosis were evaluated using H&E technique, Bax and Bcl-2 immunohistochemistry. Results suggested that betaine caused oxidative stress, inflammation, inhibition of cell growth, apoptosis, and morphological alterations in DU-145 cells dose-dependently. Furthermore, treatments with increasing betaine concentrations decreased the antioxidant levels in cells. We actually revealed that betaine, known as an antioxidant, may prevent cell proliferation by acting as an oxidant in certain doses. In conclusion, betaine may act as a biological response modifier in prostate cancer treatment in a dose-dependent manner.

Objective: Acrylamide is a chemical utilized in various industries, and many studies have demonstrated its toxicity. The NIH/3T3 mouse embryonic cell line is the standard cell line of fibroblasts, which have a pivotal role with their versatile functions in the body. However, only two studies have attempted to investigate the effect of acrylamide on these crucial cells. To fill this knowledge gap, we aimed to determine the effects of acrylamide on NIH/3T3 cells. Method: First, we performed the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay and calculated the IC₅₀ dose of acrylamide. Then, we treated cells with the IC₅₀ dose of acrylamide for 24 h and determined whether the dominant death mode of NIH/3T3 cells was apoptosis or necrosis by annexin V and caspase 3/7 assays. Finally, we performed confocal microscopy and transmission electron microscope (TEM) analysis for observing the morphological alterations. Results: MTT assay results showed that acrylamide treatment reduced the viability of NIH/3T3 cells dose-dependently and that the IC₅₀ of acrylamide was 6.73 mM. Based on annexin V and caspase 3/7 assays, the dominant death mode of NIH/3T3 cells was determined to be apoptosis. Also, caspase 3/7 activities of the acrylamide-treated NIH/3T3 cells were three times greater than those of the untreated NIH/3T3 cells. Furthermore, we observed membrane blebbing, nuclear chromatin clumping, and cytoplasmic vacuolization in TEM analysis and apparent apoptotic bodies, nuclear fragmentations, and condensations in confocal microscopy. Conclusions: In conclusion, our results suggested that the IC₅₀ of acrylamide against NIH/3T3 cells for 24 h was 6.73 mM and that acrylamide exerted its cytotoxic and anti-proliferative effects on these cells mainly via apoptosis.

Background: Acrylamide is a very common compound even reaching up to our daily foods. It has been studied in a wealth of cell lines on which it proved to have various toxic effects. Among these cell lines, human lung adenocarcinoma cell line (A549) is one of that on which acrylamide’s toxicity has not been studied well yet. Aim: We intended to determine the half maximal inhibitory concentration (IC₅₀) dose of acrylamide and to investigate its cytotoxic, anti-proliferative and apoptotic effects on A549 cells. Methods: We determined the IC₅₀ dose by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Then, the mode of cell death was evaluated by flow cytometry using Annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. Next, we performed transmission electron microscopy (TEM) and confocal microscopy analyses for morphological alterations and apoptotic indices. Results: According to the MTT assay results, A549 cell viability decreases proportionally with increasing acrylamide concentrations and IC₅₀ for A549 was 4.6 mM for 24 h. Annexin-V FITC/PI assay results indicated that acrylamide induces apoptosis in 64% of the A549 cells. TEM and confocal microscopy analyses showed nuclear condensations, fragmentations, cytoskeleton laceration, and membrane blebbing, which are morphological characteristics of apoptosis. Conclusion: Our research suggests that acrylamide causes cytotoxic, anti-proliferative, and apoptotic effects on A549 cells at 4.6 mM IC₅₀ dose in 24 h.