Exogenous p53 upregulated modulator of apoptosis (PUMA) decreases growth of lung cancer A549 cells.

Chun-Ju Liu,Da-Ya Luo,Hui-Fang Wan,Fu-Sheng Wan,Wei-Feng Zhu,Jun-Ping Yang,Xia-Li Zhang,Xiao-Jun Yang

doi:10.7314/apjcp.2015.16.2.741

Chun-Ju Liu, Da-Ya Luo + Show 6 more

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https://doi.org/10.7314/apjcp.2015.16.2.741

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Abstract

To investigate the influence of exogenous p53 upregulated modulator of apoptosis (PUMA) expression on cell proliferation and apoptosis in human non-small cell lung cancer A549 cells and transplanted tumor cell growth in nude mice. A549 cells were divided into the following groups: control, non- carrier (NC), PUMA (transfected with pCEP4- (HA) 2-PUMA plasmid), DDP (10 μg/mL cisplatin treatment) and PUMA+DDP (transfected with pCEP4-(HA)2-PUMA plasmid and 10 μg/mL cisplatin treatment). The MTT method was used to detect the cell survival rate. Cell apoptosis rates were measured by flow cytometry, and PUMA, Bax and Bcl-2 protein expression levels were measured by Western blotting. Compared to the control group, the PUMA, DDP and PUMA+DDP groups all had significantly decreased A549 cell proliferation (p<0.01), with the largest reduction in the PUMA+DDP group. Conversely, the apoptosis rates of the three groups were significantly increased (P<0.01), and the PUMA and DDP treatments were synergistic. Moreover, Bax protein levels significantly increased (p<0.01), while Bcl-2 protein levels significantly decreased (p<0.01). Finally, both the volume and the weights of transplanted tumors were significantly reduced (p<0.01), and the inhibition ratio of the PUMA+DDP group was significantly higher than in the single DDP or PUMA groups. Exogenous PUMA effectively inhibited lung cancer A549 cell proliferation and transplanted tumor growth by increasing Bax protein levels and reducing Bcl-2 protein levels.

Highlights

One of the most common malignant tumors that threatens human health, lung cancer has the highest mortality worldwide
The greatest inhibition was in the p53 upregulated modulator of apoptosis (PUMA)+DDP group, indicating that there was a synergistic reaction between PUMA and DDP treatments. These results show that the combined application of PUMA and DDP greatly inhibited A549 cell proliferation
The apoptosis rate of the NC group was not different from the control group (p>0.05). These results indicate that PUMA gene transfection promoted A549 cell apoptosis, and that even more apoptosis resulted from combined PUMA and DDP treatment

Summary

Introduction

One of the most common malignant tumors that threatens human health, lung cancer has the highest mortality worldwide. In China, lung cancer ranks second, or even first in some cities, as the cause for malignant tumor death (Zaffaroni et al, 2007; Watanabe et al, 2009; Rouibaa et al, 2013). It has been reported that overexpression of the anti-apoptotic gene Bcl-2 and decreased expression of the pro-apoptotic gene Bax could inhibit cellular apoptosis, promoting the important mechanisms of tumor development - cell survival and proliferation (Meng et al, 2010; Perez et al, 2013). It is important to study Bcl-2 and Bax gene expression in pre-cancerous pathological changes for early lung cancer diagnoses

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