Smokeless tobacco extract inhibits proliferation and promotes apoptosis in oral mucous fibroblasts

Li,Lei; Zhou,Xiaoqing; Wang,Yanran

doi:10.3892/ol.2018.9252

Smokeless tobacco extract inhibits proliferation and promotes apoptosis in oral mucous fibroblasts

Authors:
- Lei Li
- Xiaoqing Zhou
- Yanran Wang
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Affiliations: Department of Stomatology, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China, Department of Oral and Maxillofacial Surgery, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China
Published online on: August 2, 2018 https://doi.org/10.3892/ol.2018.9252
Pages: 5066-5074
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The consumption of smokeless tobacco extract (STE) is growing rapidly, and it has been implicated in several human diseases including diabetes, inflammation and a number of types of cancer. The toxicity of STE requires evaluation, as it is known to induce numerous public health issues. To investigate whether STE serves a role in cultured human oral mucosa fibroblasts (hOMFs), the present study examined HOMF morphology with inverted microscopy and immunofluorescence staining. The cell viability was measured with MTT assays, which detected the cell apoptosis rate via flow cytometry. The activities of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) were measured via flow cytometry and commercial kits, subsequent to exposing the cells to various concentrations of STE. Reverse transcription quantitative polymerase chain reaction and western blot analyses were used to demonstrate that the mRNA and the protein expression levels of cell cycle‑associated genes (cyclin‑dependent kinase inhibitor 1 and cyclin D1), apoptosis‑associated genes [B cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associatied X protein], tumor protein (p53), nuclear factor kappa light chain enhancer of activated B cells (NF‑κB)‑transcription factor (p65) signaling pathways, NF‑E2‑related factor 2 (Nrf2), heme oxygenase‑1 (HO‑1) and NAD(P)H: quinoneoxidoreductase1 (NQO1).The results indicated that the hOMF cells were positive for cytokeratin staining. STE induced G1‑S cell cycle progression and cell apoptosis by regulating the cell cycle or apoptosis‑associated proteins. STE treatment increased the concentrations of ROS and MDA, and decreased the concentrations of SOD and CAT. STE unregulated phosphorylated‑p53, NF‑κB p65, Nrf2, HO‑1, and NQO1 expression levels in the hOMF cells. The present study demonstrated that STE appears to promote oral disease.

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