HPV16 E6 gene polymorphisms and the functions of the mutation site in cervical cancer among Uygur ethnic and Han nationality women in Xinjiang, China

Huizhen Xin,Dongmei Li,Weinan Zheng,Haichen Long,Hongtao Li,Chunhe Zhang,Zemin Pan,Zhenzhen Pan,Xiangyi Zhe,Renfu Shao

doi:10.1186/s12935-022-02506-0

Abstract

BackgroundTo investigate the genotype distribution of human papillomavirus (HPV) in infected Uygur and Han women in Xinjiang, China; analyze the HPV16 E6 gene polymorphism site and relationship with the development of cervical cancer.MethodsThe HPV16 E6 sequence was analyzed using the European standard prototype to perform an evolutionary tree. HPV16 E6-T295/T350, G295/G350, and T295/G350 GV230 vectors were stably transfected into cervical cancer C33A cells to analyze the cell proliferation, migration and invasion, apoptosis by CCK8 and clonogenic assays, transwell and cell scratch assays, FACS experiments.ResultsThe total HPV infection rate was 26.390% (760/2879), whereas the Uygur 22.87% (196/857) and the Han was 27.89% (564/2022) (P < 0.05). Among 110 mutations, 65 cases of E6 genes were mutated at nucleotide 350 (T350G) with the leucine changing to valine (L83V). Moreover, there were 7 cases of E6 gene mutated at nucleotide 295 (T295G) with aspartic changing to glutamic (D64E). When E6 vector(s) of mutations sites were transfected into C33A cells, they were found to promote cellular proliferation, migration, invasion, and inhibit apoptosis. T295/G350-E6 was significantly stronger than G295/G350 and T295/T350, G295/G350 was significantly stronger than T295/T350 (P < 0.05). The T295/G350 had the strongest effect on C33A cells and G295/G350 was significantly stronger than T295/T350 (P < 0.05).ConclusionsThe positive HPV infection rates differed between the Uygur and Han in Xinjiang, China, and the genotype distribution of infection was different. After transfecting C33A cells with different eukaryotic expression vectors, the T295/G350 mutation site promoted the proliferation, migration, and invasion of C33A cells to a greater extent than G295/G350; however, G295/G350 had a stronger effect than T295/T350.

Highlights

To investigate the genotype distribution of human papillomavirus (HPV) in infected Uygur and Han women in Xinjiang, China; analyze the HPV16 Early stage gene 6 (E6) gene polymorphism site and relationship with the development of cervical cancer
HPV16 E6 gene sequencing and polymorphism analysis The full-length sequence of the HPV16 E6 gene was determined in 110 HPV16-positive patients
Expression of HPV16 E6 in cervical cancer C33A cells Asian strain (As) GV230 empty vector, HPV16 E6-T295/T350 prototype vector, HPV16 E6-G295/G350 mutation vector, and HPV16 E6-T295/G350 mutation vector were transfected into cervical cancer C33A cells, and the expression of HPV16 E6 was observed with a microscope after 48 h

Summary

Introduction

To investigate the genotype distribution of human papillomavirus (HPV) in infected Uygur and Han women in Xinjiang, China; analyze the HPV16 E6 gene polymorphism site and relationship with the development of cervical cancer. More than 20 of the detected HPV types are classified as high-risk HPV, of which HPV16 is most closely related to cervical cancer. The two most common high-risk types are HPV16 and HPV18, which cause about 70% of all cervical cancer. Cervical basal cell differentiation is a signal for HPV to transition from a latent infection to a productive infection [2]. During this process, the host cell genome integrates with the HPV genome and releases the progeny virus, resulting in a persistent HPV infection. HPV16 is the most common highrisk HPV responsible for causing cervical cancer, the mechanism by which it promotes cervical cancer is under investigation

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