Lichtenfels Martina, Almeida Lima Nunes Rafaella, Mendoza López Rossana Veronica, Alves da Silva Camila, Zeferino Luiz Carlos, de Souza Lino Vanesca, Longatto-Filho Adhemar, De Brot Louise, Rabelo-Santos Silvia Helena, Cornelio Daniela Baumann, Boccardo Enrique, de Farias Caroline Brunetto, Termini Lara
Ziel Biosciences Porto AlegreRio Grande do Sul Brazil Ziel Biosciences, Porto Alegre, Rio Grande do Sul, Brazil.
Universidade de Sao Paulo Faculdade de Medicina Hospital das Clinicas HCFMUSP São Paulo Brazil Center for Translational Research in Oncology (LIM/24), Instituto do Cancer do Estado de Sao Paulo, Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, São Paulo, Brazil.
Rev Bras Ginecol Obstet. 2025 Mar 17;47. doi: 10.61622/rbgo/2025rbgo4. eCollection 2025.
This study aimed to verify the relation between gastrin-releasing peptide receptor (GRPR), oncogenic Human Papillomavirus (HPV) and cervical lesions severity.
GRPR mRNA levels were evaluated in cervical cancer-derived cell lines and in primary keratinocytes expressing HPV16 oncogenes by RT-PCR. GRPR protein expression was assessed by immunohistochemistry in organotypic cell cultures derived from keratinocytes transduced with HPV16 oncogenes and in 208 cervical samples, including 59 non-neoplastic tissue, 28 cervical intraepithelial neoplasia grade 3 (CIN3), 44 squamous cell carcinomas (SCC) and 77 adenocarcinomas (ADC). Generic primers (GP5+/GP6+) were used to identify HPV infection in tissue samples. Experiments involving cell lines were analyzed through non-parametric tests (Kruskal Wallis), and Fisher's Exact Test for human tissues samples. All statistical tests were considered significant at p <0.05. Immunohistochemical evaluation was conducted independently and blindly by two observers (AD- LO). Any discordant findings were resolved through discussion to reach a consensus score.
GRPR mRNA levels were not increased in cells expressing HPV16 or HPV18 oncogenes. However, at the protein level, GRPR was upregulated in organotypic cell cultures containing HPV oncogenes. Besides, it was identified an association between GRPR expression and cervical lesion severity (p < 0.0001). The detection rate of high-risk HPV DNA was directly correlated with cervical disease. Nonetheless, HPV infection was not directly associated with GRPR in cervical samples.
GRPR expression is highly predictive of cervical lesion severity, irrespective of HPV infection and might contribute to improving patient's therapeutic management as well as being used a marker of disease progression.
本研究旨在验证胃泌素释放肽受体（GRPR）、致癌性人乳头瘤病毒（HPV）与宫颈病变严重程度之间的关系。
通过逆转录聚合酶链反应（RT-PCR）评估宫颈癌衍生细胞系和表达HPV16癌基因的原代角质形成细胞中GRPR mRNA水平。采用免疫组织化学法评估GRPR蛋白表达，检测对象包括转导HPV16癌基因的角质形成细胞来源的器官型细胞培养物以及208份宫颈样本，其中包括59份非肿瘤组织、28份宫颈上皮内瘤变3级（CIN3）、44份鳞状细胞癌（SCC）和77份腺癌（ADC）。使用通用引物（GP5+/GP6+）鉴定组织样本中的HPV感染。涉及细胞系的实验通过非参数检验（Kruskal Wallis）进行分析，人体组织样本采用Fisher精确检验。所有统计学检验以p<0.05为有统计学意义。免疫组织化学评估由两名观察者（AD-LO）独立且盲法进行。任何不一致的结果都通过讨论解决，以达成共识评分。
表达HPV16或HPV18癌基因的细胞中GRPR mRNA水平未升高。然而，在蛋白水平上，含有HPV癌基因的器官型细胞培养物中GRPR上调。此外，还发现GRPR表达与宫颈病变严重程度之间存在关联（p<0.0001）。高危HPV DNA的检出率与宫颈疾病直接相关。尽管如此，宫颈样本中HPV感染与GRPR无直接关联。
无论HPV感染情况如何，GRPR表达都高度预示宫颈病变的严重程度，可能有助于改善患者的治疗管理，并可作为疾病进展的标志物。
