Department of Vascular Surgery, Nanyang Central Hospital, Nanyang, Henan Province, China.
Bioengineered. 2022 Mar;13(3):6188-6195. doi: 10.1080/21655979.2022.2027064.
Long non-coding RNA (lncRNA) breast cancer-associated transcript 54 (BRCAT54) and microRNA-1269b (miR-1269b) are two critical ncRNAs in cancer biology, while their roles in hemangioma are unknown. Our preliminary sequencing data revealed their altered expression in hemangioma and predicted they could interact with each other. This study was therefore carried out to investigate the roles of BRCAT54 and miR-1269b in hemangioma, with a focus on their interaction. In this study, hemangioma samples donated by 20 infantile hemangioma patients at proliferating-phase and 20 infantile hemangioma patients at involuting-phase were used. The expression of BRCAT54 and miR-1269b in hemangioma samples, as well as hemangioma-derived endothelial cells (HDECs) and human umbilical vein endothelial cells (HUVECs) were detected by RT-qPCR. IntaRNA 2.0 was applied to predict the interaction between BRCAT54 and miR-1269b, which was then confirmed by RNA-RNA pulldown assay. Accumulation of BRCAT54 in the subcellular location of HDECs was detected by subcellular fractionation assay. The role of BRCAT54 and miR-1269b in cell proliferation has been explored by the BrdU assay. Compared to proliferating-phase tissues, involuting-phase tissues exhibited decreased expression levels of BRCAT54 and increased expression levels of miR-1269b. HDECs had decreased expression levels of BRCAT54 and increased expression levels of miR-1269b compared to that of HUVECs. In HDECs, BRCAT54, which was detected in both nuclear and cytoplasm fractions, directly interacted with miR-1269b. BRCAT54 and miR-1269b did not affect the expression of each other, while BRCAT54 suppressed the role of miR-1269b in enhancing the proliferation of HDECs. BRCAT54 may sponge miR-1269b to suppress the proliferation of HDECs.
长链非编码 RNA（lncRNA）乳腺癌相关转录物 54（BRCAT54）和 microRNA-1269b（miR-1269b）是癌症生物学中两种重要的非编码 RNA，但其在血管瘤中的作用尚不清楚。我们的初步测序数据显示它们在血管瘤中的表达发生了改变，并预测它们可能相互作用。因此，本研究旨在研究 BRCAT54 和 miR-1269b 在血管瘤中的作用，重点研究它们的相互作用。本研究使用了 20 例增殖期婴儿血管瘤患者和 20 例消退期婴儿血管瘤患者捐赠的血管瘤样本。通过 RT-qPCR 检测血管瘤样本、血管瘤衍生的内皮细胞（HDECs）和人脐静脉内皮细胞（HUVECs）中 BRCAT54 和 miR-1269b 的表达。应用 IntaRNA 2.0 预测 BRCAT54 和 miR-1269b 之间的相互作用，然后通过 RNA-RNA 下拉实验进行验证。通过亚细胞分离实验检测 BRCAT54 在 HDECs 亚细胞位置的积累。通过 BrdU 实验探讨了 BRCAT54 和 miR-1269b 在细胞增殖中的作用。与增殖期组织相比，消退期组织中 BRCAT54 的表达水平降低，miR-1269b 的表达水平升高。与 HUVECs 相比，HDECs 中 BRCAT54 的表达水平降低，miR-1269b 的表达水平升高。在 HDECs 中，BRCAT54 被检测到存在于核和细胞质部分，直接与 miR-1269b 相互作用。BRCAT54 和 miR-1269b 彼此不影响对方的表达，而 BRCAT54 抑制了 miR-1269b 增强 HDECs 增殖的作用。BRCAT54 可能通过海绵吸附 miR-1269b 抑制 HDECs 的增殖。
