EPB41L4A-AS1 regulates cervical cancer by proliferative cells: mendelian randomization and single-cell transcriptomics analyses.

Background: The current literature lacks reports on the roles of proliferative cells in tumorigenesis and causal relationship between proliferative cells and cervical cancer. This study aims to investigate the role and mechanism of proliferative cells in cervical cancer. Methods: Single-cell transcriptomics of cervical cancer were utilized to identify proliferative cells. Mendelian randomization (MR) and meta-analysis were employed to study the causal relationship between proliferative cells and cervical cancer. Additional assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry, gene set enrichment analysis (GSEA), and weighted gene co-expression network analysis (WGCNA) were exploited to study function of EPB41L4A-AS1 in the regulation of cell proliferation. Both complementary DNA (cDNA) microarray and GSEA were performed to elucidate the underlying mechanisms by which EPB41L4A-AS1 influenced proliferative cells. Results: Cervical cancer exhibited a higher proportion of proliferative cells in tumor tissue compared to healthy tissue, as evidenced by single-cell transcriptomics. Genes specifically expressed in proliferative cells were found to be predictive of the prognosis of cervical cancer patients [P=0.009; hazard ratio (high groups) =1.893; 95% confidence interval: 1.169-3.064]. Proliferative cells, rather than squamous or columnar epithelial cells, were causally associated with cervical cancer. Mechanistically, EPB41L4A-AS1 was found to regulate proliferative cells (P<0.005), described as EPB41L4A-AS1-regulated genes which were predominantly enriched in proliferative cells. The mapping of pathways associated with EPB41L4A-AS1-regulated genes to proliferative cells revealed a significant enrichment of mitosis-related pathways (normalized enrichment score >1). Furthermore, knockdown of EPB41L4A-AS1 resulted in an increased number of cells during the M phase (Sh-NC: 2N: 74.5%, S: 11.7%, 4N: 10.0%; Sh-EPB41L4A-AS1: 2N: 66.0%, S: 11.2%, 4N: 18.7%), thereby promoting cell proliferation. Conclusions: This study offered a novel perspective on the role of EPB41L4A-AS1 in regulating cervical cancer through its impact on proliferative cells.