Model of Human Tongue Squamous Cell Lines Stably Transfected with Human Papillomavirus (HPV)16 E6 and E7 Genes and Biological Characteristic Analysis.

Zilian Lan,Hengyuan Guo,Zhaoshou Yang,Zifan Yang,Xuan Pan,Ziyao Jia,Junyan Liu

doi:10.1155/2021/9968691

Abstract

Background Human tongue squamous cell carcinoma (TSCC) is the most common oral cancer with the highest human papillomavirus (HPV) infection rate in oral cancer. The purpose of this study was to research the correlation between HPV and TSCC. Method Plasmid pEGFP/HPV16 E6E7 and plasmid pEGFP/no HPV16 E6E7 were constructed. TSCC cell lines SCC9 and SCC15 were infected by liposome transfection and would be highly selected by antibiotic. Fluorescence imaging, PCR, and Western blot were used to detect the expression of HPV16 E6E7 in cells. The biological characteristics were detected by CCK-8, wound healing assay, qRT-PCR, and Western blot. Result TSCC cell lines transfected with HPV16 E6E7 gene were successfully established and identified. And the proliferation and migration ability of the TSCC cell lines infected with HPV16 E6E7 gene were significantly stronger than that of the blank group. Conclusion TSCC cell lines infected with HPV16 E6E7 with significantly higher ability of proliferation and migration were more malignant than those not infected with HPV16 E6E7.

Highlights

Oral cancer is the sixth most common cancer in the world, with high morbidity and mortality, accounting for more than 50% of confirmed patients, among which human tongue squamous cell carcinoma (TSCC) is the most common oral cancer [1]
The target sequence inserted into the plasmid vector which was designed for high expression of the fusion gene HPV16E6E7 was detected by the application of gene sequence analysis
SCC9 transfected with the pEGFP/no HPV16 E6E7 SCC9 transfected with the pEGFP/HPV16 E6E7 SCC15 transfected with the pEGFP/no HPV16 E6E7 SCC15 transfected with the pEGFP/ HPV16 E6E7

Summary

Introduction

Oral cancer is the sixth most common cancer in the world, with high morbidity and mortality, accounting for more than 50% of confirmed patients, among which human tongue squamous cell carcinoma (TSCC) is the most common oral cancer [1]. The evercreasing clinical epidemiological and experimental evidence in recent decades shows that oral cancer has close relationships with sexual behavior such as oral-vagina, oral-anal, and oral-penis [7,8,9]. It is a comfortable and encouraging finding that HPV-associated OSCC have better clinical prognosis: the reported 3-year and 5-year OS rates even, respectively, increased to 93% and 89%, which means HPV has a BioMed Research International promising effect on improving the prognosis of patients with oral squamous cell carcinomas [10, 11]. TSCC cell lines infected with HPV16 E6E7 with significantly higher ability of proliferation and migration were more malignant than those not infected with HPV16 E6E7

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