HNRNPA2/B1 is upregulated in endocrine-resistant LCC9 breast cancer cells and alters the miRNA transcriptome when overexpressed in MCF-7 cells
HNRNPA2/B1 is upregulated in endocrine-resistant LCC9 breast cancer cells and alters the miRNA transcriptome when overexpressed in MCF-7 cells
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DOI：
10.1038/s41598-019-45636-8
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发表时间：
2019-07-01
影响因子：
4.6
通讯作者：
Rouchka, Eric C.
中科院分区：
文献类型：
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作者：
Klinge, Carolyn M.;Piell, Kellianne M.;Rouchka, Eric C.
MicroRNA 在乳腺癌中失调。异质核核糖核蛋白 A2/B1 (HNRNPA2/B1) 是初级 miRNA (pri-miRNA) 中 N(6)-甲基腺苷 (m6A) 标记的读取器，并促进 DROSHA 加工为前体 miRNA (pre-miRNA)。我们检查了 m6A 的写入器、读取器和擦除器的表达，并报告说，与亲本、他莫昔芬敏感的 MCF-7 细胞相比，HNRNPA2/B1 在他莫昔芬耐药的 LCC9 乳腺癌细胞中表达更高。为了研究 HNRNPA2/B1 表达的增加如何影响 miRNA 表达，HNRNPA2/B1 在 MCF-7 细胞中瞬时过表达（大约 5.4 倍），以进行全基因组 miRNA 分析 (miRNA-seq)。转染后48小时，分别有148和88个miRNA上调和下调；转染后72小时，分别上调和下调177和172个miRNA。 MetaCore 富集分析确定了乳腺癌中黄体酮受体作用和通过 miRNA 进行的转化生长因子 β (TGF β) 信号传导是上调 miRNA 的下游途径，而通过 SMAD 和 Notch 信号传导的 TGF β 信号传导是下调 miRNA 的途径。 HNRNPA2/B1 调节的 miRNA 的 mRNA 靶标的 GO 生物学过程包括对雌二醇和细胞基质粘附的反应。 qPCR 证实 HNRNPA2B1 下调 miR-29a-3p、miR-29b-3p 和 miR-222，上调 miR-1266-5p、miR-1268a、miR-671-3p。 HNRNPA2/B1 的短暂过度表达降低了 MCF-7 对 4-羟基他莫昔芬和氟维司群的敏感性，表明 HNRNPA2/B1 在内分泌抵抗中的作用。
MicroRNAs are dysregulated in breast cancer. Heterogeneous Nuclear Ribonucleoprotein A2/B1 (HNRNPA2/B1) is a reader of the N(6)-methyladenosine (m6A) mark in primary-miRNAs (pri-miRNAs) and promotes DROSHA processing to precursor-miRNAs (pre-miRNAs). We examined the expression of writers, readers, and erasers of m6A and report that HNRNPA2/B1 expression is higher in tamoxifen-resistant LCC9 breast cancer cells as compared to parental, tamoxifen-sensitive MCF-7 cells. To examine how increased expression of HNRNPA2/B1 affects miRNA expression, HNRNPA2/B1 was transiently overexpressed (similar to 5.4-fold) in MCF-7 cells for whole genome miRNA profiling (miRNA-seq). 148 and 88 miRNAs were up- and down-regulated, respectively, 48 h after transfection and 177 and 172 up- and down-regulated, respectively, 72 h after transfection. MetaCore Enrichment analysis identified progesterone receptor action and transforming growth factor beta (TGF beta) signaling via miRNA in breast cancer as pathways downstream of the upregulated miRNAs and TGF beta signaling via SMADs and Notch signaling as pathways of the downregulated miRNAs. GO biological processes for mRNA targets of HNRNPA2/B1-regulated miRNAs included response to estradiol and cell-substrate adhesion. qPCR confirmed HNRNPA2B1 downregulation of miR-29a-3p, miR-29b-3p, and miR-222 and upregulation of miR-1266-5p, miR-1268a, miR-671-3p. Transient overexpression of HNRNPA2/B1 reduced MCF-7 sensitivity to 4-hydroxytamoxifen and fulvestrant, suggesting a role for HNRNPA2/B1 in endocrine-resistance.