Sasivimolrattana Thanayod, Gunawan Aileen, Wattanathavorn Warattaya, Pholpong Chavis, Chaiwongkot Arkom, Bhattarakosol Pattarasinee, Bhattarakosol Parvapan
Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand.
Center of Excellence in Applied Medical Virology, Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
PeerJ. 2024 Dec 6;12:e18601. doi: 10.7717/peerj.18601. eCollection 2024.
Cervical cancer remains a significant global health concern, ranking as the fourth most prevalent cancer among women worldwide. Human papillomaviruses (HPV) transcribe many genes that might be responsible for cervical cancer development. This study aims to investigate the correlation between the expression of HPV16 early genes and the mRNA expression of human FOXO3a, a tumor suppressor gene, in association with various stages of cervical precancerous lesions.
Eighty-five positive HPV16 DNA cervical swab samples were recruited and categorized based on cytology stages, , negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cell cannot exclude HSIL (ASC-H), high-grade squamous intraepithelial lesion (HSIL). RT-qPCR was performed to amplify HPV16E1, E4, E6, E6*I, E7, and human FOXO3a mRNA expression in all samples. The relative expression of those genes was calculated using GAPDH as a control. Detection of FOXO3a mRNA expression in the cervical cancer cell line by RT-qPCR and meta-analysis of FOXO3a expression using the RNA-Seq dataset by GEPIA2 were analyzed to support the conclusions.
Among the cervical samples, HPV16E1 and E7 were significantly increased expression correlating to disease severity. HPV16E4 mRNA expression was 100% detected in all LSIL samples, with a significant increase observed from normal to LSIL stages. Conversely, FOXO3a mRNA expression decreased with disease severity, and the lowest expression was observed in HSIL/squamous cell carcinoma (SCC) samples. In addition, similar results of FOXO3a downregulation were also found in the cervical cancer cell line and RNA-Seq dataset of cervical cancer samples.
HPV16 early mRNA levels, including E1 and E7, increase during cancer progression, and downregulation of FOXO3a mRNA is a characteristic of cervical cancer cells and HSIL/SCC. Additionally, HPV16E4 mRNA expression was consistently detected in all LSIL samples, suggesting the presence of active viral replication. These findings might lead to further investigation into the interplay between HPV gene expression and host cell factors for targeted therapeutic strategies in cervical cancer management.
宫颈癌仍然是一个重大的全球健康问题，在全球女性中是第四大最常见的癌症。人乳头瘤病毒（HPV）转录许多可能与宫颈癌发展相关的基因。本研究旨在探讨HPV16早期基因的表达与肿瘤抑制基因人FOXO3a的mRNA表达之间的相关性，以及与宫颈上皮内瘤变各阶段的关系。
收集85份HPV16 DNA阳性的宫颈拭子样本，并根据细胞学阶段进行分类，即上皮内病变或恶性肿瘤阴性（NILM）、意义不明确的非典型鳞状细胞（ASC-US）、低级别鳞状上皮内病变（LSIL）、不能排除高级别鳞状上皮内病变的非典型鳞状细胞（ASC-H）、高级别鳞状上皮内病变（HSIL）。对所有样本进行RT-qPCR，以扩增HPV16 E1、E4、E6、E6*I、E7和人FOXO3a的mRNA表达。以GAPDH作为对照计算这些基因的相对表达。通过RT-qPCR检测宫颈癌细胞系中FOXO3a mRNA的表达，并使用GEPIA2通过RNA-Seq数据集对FOXO3a的表达进行荟萃分析，以支持研究结论。
在宫颈样本中，HPV16 E1和E7的表达与疾病严重程度显著相关且明显增加。在所有LSIL样本中均检测到HPV16 E4 mRNA表达，从正常阶段到LSIL阶段观察到显著增加。相反，FOXO3a mRNA表达随疾病严重程度降低，在HSIL/鳞状细胞癌（SCC）样本中观察到最低表达。此外，在宫颈癌细胞系和宫颈癌样本的RNA-Seq数据集中也发现了FOXO3a下调的类似结果。
HPV16早期mRNA水平，包括E1和E7，在癌症进展过程中升高，FOXO3a mRNA下调是宫颈癌细胞和HSIL/SCC的一个特征。此外，在所有LSIL样本中均持续检测到HPV16 E4 mRNA表达，表明存在活跃的病毒复制。这些发现可能会促使进一步研究HPV基因表达与宿主细胞因子之间的相互作用，以制定宫颈癌管理的靶向治疗策略。
