REST/NRSF Silencing Modifies Neuronal Gene Expression in siRNA-Treated HeLa Cells - A Preliminary Exploration in the Search for Neuronal Biomarkers of Cervical Cancer

Sep 24, 2026

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REST (RE1-silencing transcription factor) diminution is linked to transcriptional relaxation, neuropeptide overexpression, and phenotype redefinition in neuroendocrine cancers, yet its impact on cervical cancer remains insufficiently characterized. This work evaluates neuronal gene expression in cancerous cells with reduced REST using tissue samples from southern Mexico, immunostaining of REST, in silico target selection, and RT-PCR in siRNA-treated HeLa cells. Results show REST reduction in premalignant lesions, SCC, ADC, and cancer cell lines, with altered RE1-containing gene expression (e.g., CgA, CHRNB2, BDNF, CRF, RASSF1A), supporting preliminary biomarker potential.

LanguageEnglish

CategoryHealthcare

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Pages8

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Abstract

Background and Objectives

Materials and Methods

Results

Conclusions

Introduction

medicina Communication REST/NRSF Silencing Modiﬁes Neuronal Gene Expression in siRNA-Treated HeLa Cells: A Preliminary Exploration in the Search for Neuronal Biomarkers of Cervical Cancer Karen Cort²s-Sarabia 1, Luz Del Carmen Alarcân-Romero 1, Miguel 􀂁ngel Mendoza-Catal¡n 1, Juan Carlos Carpio-Pedroza 2, Eduardo Castañeda-Saucedo 1 and Carlos Ortuño-Pineda 1, * Citation: Cortés-Sarabia, K.; Alarcón-Romero, L.D.C.; Mendoza-Catalán, M.Á.; Carpio-Pedroza, J.C.; Castañeda-Saucedo, E.; Ortuño-Pineda, C. REST/NRSF Silencing Modiﬁes Neuronal Gene Expression in siRNA-Treated HeLa Cells: A Preliminary Exploration in the Search for Neuronal Biomarkers of Cervical Cancer. Medicina 2023, 59, 537. [https://doi.org/10.3390/](https://doi.org/10.3390/)[ ](https://doi.org/10.3390/)medicina59030537 Academic Editor: Konstantinos Dimas Received: 13 December 2022 Revised: 3 March 2023 Accepted: 6 March 2023 Published: 9 March 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) . 1 Facultad de Ciencias Qu½mico-Biolâgicas, Universidad Autânoma de Guerrero, Av. L¡zaro C¡rdenas s/n, Chilpancingo 39090, Mexico 2 Departamento de Parasitolog½a, Instituto de Diagnâstico y Referencia Epidemiolâgicos “Dr. Manuel Mart½nez B¡ez”, Francisco de P. Miranda 177, Mexico City 01480, Mexico * Correspondence: [ortunoc@outlook.com](ortunoc@outlook.com); Tel.: +52-55-3154-3508 Abstract: Background and Objectives: REST (RE1-silencing transcription factor) diminution is associated with transcriptional relaxation, neuropeptide overexpression, and phenotype redeﬁnition inneuroendocrine cancers, but this effect has barely been studied in cervical cancer (CC). We previously reported reduced expressions of REST in samples with premalignant lesions and CC; however, the transcriptional consequences for neural genes associated with reduced REST expression in CC are unknown. Therefore, the objective of this work was to evaluate the expression of neuronal genes in cancerous cells with reduced expression levels of REST. Materials and Methods: Here, we monitored levels of REST by immunostaining along the premalignant lesions and in invasive cervical squamous cell carcinoma (SCC) and endocervical adenocarcinoma (ADC) in tissue samples from female patients from southern Mexico and the derivative cell lines SiHa and HeLa, respectively. Next, we selected REST target genes in silico and explored the effect of REST silencing by RT-PCR in siRNA-treated HeLa cells. Results: The results show a REST diminution in premalignant lesions, SCC, ADC, and cancerous cell lines. Further REST silencing in HeLa cells altered the expression of genes containing the RE1 (Restrictive Element 1) sequence, including CgA (chromogranin A), CHRN􀀌2 (cholinergic receptor nicotinic 􀀌 2 subunit), BDNF (brain-derived neurotrophic factor), CRF (corticotropin-releasing factor), and RASSF1A (Ras association domain family 1) . Conclusions: This work provides preliminary evidence of the role of REST loss in the transcriptional regulation of its target genes in HeLa cells, which could have positive implications for the search for new biomarkers of cervical cancer. Keywords: neural phenotype; REST targets; transcriptional regulation; HeLa cells; cervical intraepithelial neoplasia 1. Introduction In the epithelial tissues of mammals, REST (also known as neuron-restrictive silencer factor, NRSF) binds to targets of the RE1 (Restrictive element 1) sequence and represses neural gene transcription by orchestrating chromatin inactivation. In contrast, the degradation of REST promotes neurogenesis via transcriptional relaxation during embryogenesis and neuronal phenotype maintenance

FAQs

What is the study objective regarding REST in cervical cancer?

To evaluate neuronal gene expression in cancerous cells with reduced REST levels, addressing transcriptional consequences relevant to cervical cancer biomarker discovery.

How were REST levels monitored and which models were used?

REST was monitored by immunostaining in premalignant lesions and invasive cervical SCC and ADC tissue samples, and in corresponding derivative cell lines SiHa and HeLa.

Which neuronal genes changed after REST silencing in siRNA-treated HeLa cells?

REST silencing altered expression of RE1-containing genes including CgA, CHRNB2, BDNF, CRF, and RASSF1A.

Sep 24, 2026

Uploaded by Logic