Effects of HPV-16 infection on hypopharyngeal squamous cell carcinoma and FaDu cells

Abstract

Hypopharyngeal squamous cell carcinoma is a common type of malignant tumor among head and neck squamous cell carcinomas (HNSCCs). Heavy smoking and/or drinking is associated with the development of HNSCC. However, HNSCC also occurs in individuals that do not drink or smoke, possibly due to infection with the human papilloma virus (HPV). HPV-16 has been shown to be closely associated with the occurrence of several types of cancers. However, its role in hypopharyngeal squamous cell carcinoma remains unclear. In the present study, we investigated the effects of HPV-16 on hypopharyngeal squamous cell carcinoma and FaDu cells. Lentiviral vectors were used to establish FaDu cells that expressed the E6 and E7 proteins of HPV-16. We used quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays and western blotting to detect and determine the levels of expression for E6-E7 mRNAs and proteins. Cell Counting Kit-8 (CCK-8) assays, enzyme-linked immunosorbent assays (ELISA), Transwell assays, and flow cytometry were used to assess the effects of HPV-16 E6-E7 on the proliferation, invasion, metastasis and apoptosis of FaDu cells. Expression of microRNAs was analyzed by qRT-PCR. We found that the expression levels of HPV-16 E6-E7 were increased in FaDu cells transfected with the lentiviral vector compared with that observed in the control cells. In addition, the rates of apoptosis were decreased in the transfected cells, while proliferation was increased. The average numbers of cells penetrating the Matrigel were significantly higher than those for the controls. We detected miR-363 and miR-15a, and their expression levels were significantly increased in the HPV-16-positive patients and in FaDu cells expressing HPV-16 E6-E7. We found that HPV-16 E6-E7 appeared to inhibit apoptosis, and to increase cell proliferation, invasion and metastasis. Furthermore, miR-363 and miR-15a were overexpressed in the hypopharyngeal squamous cell carcinoma samples infected with HPV-16, and in FaDu cells stably expressing HPV-16 E6-E7. These findings may provide a new clue of the mechanisms involved in the pathogenesis of HPV-16-positive hypopharyngeal squamous cell carcinoma.

miR‑490‑5p regulates the proliferation, migration, invasion and epithelial‑mesenchymal transition of pharyngolaryngeal cancer cells by targeting mitogen‑activated protein kinase kinasekinase 9

“…miR-490-5p expression was also detected in NP69, HaCaT, BICR 18, FaDu, HNE-3 and Detroit 562 cells, and the result demonstrated that miR-490-5p expression in pharyngolaryngeal cancer cells was decreased, particularly in BICR 18 and FaDu cells, which further confirmed that the expression of miR-490-5p was downregulated in pharyngolaryngeal cancer cells. In addition, laryngeal carcinoma is one of the most common head and neck neoplasms, which is mainly represented by squamous cell carcinoma was predominant, BICR 18 and FaDu cells are squamous cells (20,21). Therefore, BICR 18 and FaDu cells were selected for later experiments.…”

miR-15a induces cell apoptosis by targeting BCL2L2 and BCL2 in HPV-positive hypopharyngeal squamous cell carcinoma

“…In addition, several cellular miRNAs are differentially expressed in HPV-positive HNSCC cell lines and in HPV-negative HNSCC cell lines (27). In our previous study (24), we detected the expression of miR-15a in HNSCC tissues, and found that miR-15a expression was significantly higher in HPV-positive than in HPV-negative HNSCC. These results suggested that miR-15a plays an important role in HPV-positive HNSCC and acts as an anti-oncogene.…”

“…Our previous study (24) showed that miR-15a is highly expressed in HPV-positive tissues and cells. It may be speculated that HPV-positive tumors have a better prognosis.…”

A 3‐miRNA signature predicts survival of patients with hypopharyngeal squamous cell carcinoma after post‐operative radiotherapy

“…Wang found miR‐203 could inhibit tumour growth and metastasis through PDPN . MiR‐15a was up‐regulated in HPV‐positive HSCC and might induce tumour apoptosis via BCL2L2 and BCL2 …”

“…23 MiR-15a was up-regulated in HPV-positive HSCC and might induce tumour apoptosis via BCL2L2 and BCL2. 24,25 However, many miRNAs are oncogenic by targeting tumour suppressor genes. miR-21 was widely accepted as 'oncomiR', which was overexpressed in various types of cancers including lymphoma, oesophageal cancer, LSCC, HSCC and SCC of oral cavity.…”