Elsevier

Toxicology in Vitro

Volume 58, August 2019, Pages 35-41
Toxicology in Vitro

MEHP promotes the proliferation of oral cancer cells via down regulation of miR-27b-5p and miR-372-5p

https://doi.org/10.1016/j.tiv.2019.03.014Get rights and content

Highlights

  • MEHP triggers the proliferation of OSCC cells.

  • MEHP increases the expression of PCNA.

  • MEHP decreases the expression of miR-27b-5p and miR-372-5p.

  • c-Myc is involved in MEHP regulated expression of miR-27b-5p.

Abstract

Mono-2-ethyhexyl phthalate (MEHP), an environmental xenoestrogen, is widely used in the production of polyvinyl chloride materials and can be easily accumulated into human body. Emerging evidences showed that MEHP can regulate the progression of various cancers. Oral cancer cells could be directly exposed to MEHP during food and water digestion, while the roles of MEHP on the progression of oral cancer were rarely investigated. Our present study found that MEHP can trigger the proliferation of oral squamous cell carcinoma (OSCC) cells and increase the expression of proliferating cell nuclear antigen (PCNA). We checked the expression of various miRNAs which can target the 3’UTR of PCNA. Specifically, MEHP can decrease the expression of miR-27b-5p and miR-372-5p, which can directly bind with the 3’UTR of PCNA to inhibit its expression. Over expression of miR-27b-5p and miR-372-5p can abolish MEHP induced cell proliferation and expression of PCNA in OSCC cells. Further, MEHP can induce the expression of c-Myc, which can suppress the transcription of miR-27b-5p in OSCC cells. In vivo xenograft study on the basis of SCC-4 cells confirmed that MEHP can trigger the growth of OSCC and suppress the expression of miR-27b-5p and miR-372-5p. Collectively, our present study suggested that MEHP can promote the growth and progression of OSCC via down regulation of miR-27b-5p and miR-372-5p.

Introduction

Oral cancer is the most common subtype of head and neck cancers (Mascolo et al., 2012). Although oral cancers may originate in any of the tissues of the mouth, oral squamous cell carcinoma (OSCC) accounts for >90% of oral cancer (Torre et al., 2015). The 5-year survival rate of OSCC is approximately 63% (Noguti et al., 2012). The mortality of OSCC is high due to it is commonly seen in advanced stages before treatment (Saman, 2012). It has been reported that the incidence of OSCC is strongly associated with the risk factors such as alcohol consumption, tobacco abuse, chewing betel quid, and fruit and vegetable deficiency in diet (Warnakulasuriya, 2009). Therefore, the illustrations of risk factors responsible for the progression of OSCC will be great helpful for the cancer prevention.

Both epidemiological and laboratory studies indicated that estrogenic signals can regulate the progression of OSCC both in vitro and in vivo (Doll et al., 2015). It has been reported that environmental endocrine disrupting chemicals (EDCs) can regulate the progression of various cancers including OSCC (Gao et al., 2015). Di (2-ethylhexyl) phthalate (DEHP) is widely used in the production of polyvinyl chloride medical materials in our daily life, which could be absorbed into human body via food digestion and/or inhalation (Erythropel et al., 2014). It has been reported that DEHP can cause liver tumorigenesis presumably via peroxisome proliferator-activated receptor alpha (PPARalpha) dependent and independent manners (Ito et al., 2007; Voss et al., 2005). Mono-(2-ethylhexyl) phthalate (MEHP) is the most toxic metabolite of DEHP in human body (Trasande et al., 2013). It has been reported that MEHP can trigger proliferation of cervical cancer (Yang et al., 2018) and migration of Wilm's cancer via activation of NF-κB signals (Wang et al., 2017a). Although OSCC cells can be directly exposed to MEHP during food digestion or inhalation, the effect and related mechanisms of MEHP on the progression of OSCC remain unclear.

It has been reported that microRNAs (miRNAs) can bind to the 3′-untranslated region of mRNA and then regulate the progression of various cancer including OSCC (Nadeem et al., 2018). For example, miR-134 can target PDCD7 to reduce E-cadherin expression and enhance oral cancer progression (Peng et al., 2018). While miR-143 can suppress the growth and invasion of OSCC cells via targeting hexokinase 2 (Sun and Zhang, 2017). MEHP can regulate the expression of miRNAs in mammalian cells (Meruvu et al., 2016; Meruvu et al., 2016). The present study finds that MEHP can trigger the proliferation of OSCC cells and increase the expression of proliferating cell nuclear antigen (PCNA), an auxiliary protein of DNA polymerise-5 that is concentrated in the nucleus. Further, down regulation of miR-27b-5p and miR-372-5p was involved in MEHP triggered progression of OSCC.

Section snippets

Cell culture

The human OSCC SCC-4, SCC-9, and SCC-25 cells were purchased from the American Type Cell Collection (ATCC Manassas, VA, USA) and cultured in medium recommended by ATCC containing 5% heat-inactivated fetal bovine serum (FBS) and 10 μg/ml penicillin-streptomycin. Cells were maintained in a humidified incubator at 37 °C with atmosphere of 5% (v/v) CO2 in air. As to cell treatment, all plastic items and water were pretreated by enhanced sonochemical degradation to reduce the background

MEHP can trigger the proliferation of OSCC cells

In order to evaluation the potential effects of MEHP on the progression of OSCC, SCC-4, SCC-9 and SCC-25 cells were treated with 10 and 100 nM MEHP for 48 h. Our data showed that both 10 and 100 nM MEHP can trigger the proliferation of SCC-4, SCC-9 and SCC-25 cells after treated for 48 h (Fig. 1A, B and C). Further, 10 nM MEHP can trigger the proliferation of SCC-4 (Fig. 1D) and SCC-25 (Fig. 1E) cells via a time dependent manner. Western blot analysis showed that 10 nM MEHP can increase the

Discussion

Although MEHP can trigger proliferation of cervical cancer (Yang et al., 2018) and migration of Wilm's cancer via activation of NF-κB signals (Wang et al., 2017a), its effect on the oral cancer was not illustrated. Our present study found that MEHP can trigger the in vitro and in vivo growth of OSCC cells and upregulation of PCNA. This was due to that MEHP can down regulate the expression of miR-27b-5p and miR-372-5p in OSCC cells, which can directly target the 3’UTR of PCNA mRNA and then

Conflict of interest

The authors declare no conflict of interest.

Acknowledgements

This research was supported by the Health Department Issue of Heilongjiang, China (No. 2017156) and the Natural Science Foundation of Heilongjiang, China (No. QC2015124).

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