子宫内膜异位症是一种良性炎症性疾病，其特点是子宫内膜样组织在子宫外生长，是子宫内膜异位相关卵巢癌的危险因素。特别是卵巢内膜囊肿，即深层浸润性子宫内膜异位病变，被认为是卵巢透明细胞癌（OCCC）的前体病变。为了探究这种转录组景观，我们通过批量RNA和小RNA测序，比较了已被病理学证实为合并子宫内膜异位病变的OCCC（n = 4）与良性内膜囊肿（n = 4）之间的差异。蛋白质编码基因分析发现，在OCCC合并子宫内膜异位病变与内膜囊肿组织中，有2449个上调和3131个下调的蛋白质编码基因（DESeq2，P < 0.05，log2倍变化 > |1|）。基因集富集分析显示，细胞周期调节和DNA复制等通路上调，细胞因子受体信号通路和基质组下调。将临床样本与公开可获得的OCCC细胞株数据集进行通路激活分数比较，发现OCCC合并子宫内膜异位病变与OVTOKO、OVISE、RMG1、OVMANA、TOV21G、IGROV1和JHOC5细胞株之间存在显著的分子相似性。miRNA分析发现，有64个上调和61个下调的成熟miRNA分子（DESeq2，P < 0.05，log2倍变化 > |1|）。在OCCC合并子宫内膜异位病变中，miR-10a-5p代表超过21％的miRNA分子，并且显著上调（NGS：log2倍变化= 4.37，P = 2.43e-18； QPCR：8.1倍变化，P < 0.05）。体外下调miR-10a在OCCC细胞株中的表达水平与卡铂的50%抑制浓度（IC50）呈正相关（R2 = 0.93）。与非靶向控制miRNA转染相比，miR-10a在体外过表达显著降低细胞增殖（n = 6；P < 0.05）。类似地，细胞周期分析显示细胞从S期和G2期转移到G1期（n = 6；P < 0.0001）。生物信息学分析预测，OCCC合并子宫内膜异位病变中下调miR-10a-5p靶基因涉及受体信号通路、增殖和细胞周期进展。体外过表达miR-10a与预测的在增殖、细胞周期调节和细胞存活中起关键作用的miR-10a靶基因（SERPINE1，下调3倍；P < 0.05；CDK6，下调2.4倍；P < 0.05；RAP2A，下调2-3倍；P < 0.05）的表达显著降低。这些OCCC研究表明，miR-10a-5p是一种有影响力的潜在致癌分子，需要进一步研究。 版权所有 © 2023 Collins, Wang, Klymenko, Davis, Martinez, Zhang, So, Buechlein, Rusch, Creighton和Hawkins。

Endometriosis, a benign inflammatory disease whereby endometrial-like tissue grows outside the uterus, is a risk factor for endometriosis-associated ovarian cancers. In particular, ovarian endometriomas, cystic lesions of deeply invasive endometriosis, are considered the precursor lesion for ovarian clear-cell carcinoma (OCCC).To explore this transcriptomic landscape, OCCC from women with pathology-proven concurrent endometriosis (n = 4) were compared to benign endometriomas (n = 4) by bulk RNA and small-RNA sequencing.Analysis of protein-coding genes identified 2449 upregulated and 3131 downregulated protein-coding genes (DESeq2, P< 0.05, log2 fold-change > |1|) in OCCC with concurrent endometriosis compared to endometriomas. Gene set enrichment analysis showed upregulation of pathways involved in cell cycle regulation and DNA replication and downregulation of pathways involved in cytokine receptor signaling and matrisome. Comparison of pathway activation scores between the clinical samples and publicly-available datasets for OCCC cell lines revealed significant molecular similarities between OCCC with concurrent endometriosis and OVTOKO, OVISE, RMG1, OVMANA, TOV21G, IGROV1, and JHOC5 cell lines. Analysis of miRNAs revealed 64 upregulated and 61 downregulated mature miRNA molecules (DESeq2, P< 0.05, log2 fold-change > |1|). MiR-10a-5p represented over 21% of the miRNA molecules in OCCC with endometriosis and was significantly upregulated (NGS: log2fold change = 4.37, P = 2.43e-18; QPCR: 8.1-fold change, P< 0.05). Correlation between miR-10a expression level in OCCC cell lines and IC50 (50% inhibitory concentration) of carboplatin in vitro revealed a positive correlation (R2 = 0.93). MiR-10a overexpression in vitro resulted in a significant decrease in proliferation (n = 6; P< 0.05) compared to transfection with a non-targeting control miRNA. Similarly, the cell-cycle analysis revealed a significant shift in cells from S and G2 to G1 (n = 6; P< 0.0001). Bioinformatic analysis predicted that miR-10a-5p target genes that were downregulated in OCCC with endometriosis were involved in receptor signaling pathways, proliferation, and cell cycle progression. MiR-10a overexpression in vitro was correlated with decreased expression of predicted miR-10a target genes critical for proliferation, cell-cycle regulation, and cell survival including [SERPINE1 (3-fold downregulated; P< 0.05), CDK6 (2.4-fold downregulated; P< 0.05), and RAP2A (2-3-fold downregulated; P< 0.05)].These studies in OCCC suggest that miR-10a-5p is an impactful, potentially oncogenic molecule, which warrants further studies.Copyright © 2023 Collins, Wang, Klymenko, Davis, Martinez, Zhang, So, Buechlein, Rusch, Creighton and Hawkins.