Circ‐POLR3A accelerates TGF‐β2‐induced promotion in cell viability, migration, and invasion of lens epithelial cells via miR‐31/TXNIP signaling cascade
Circ‐POLR3A accelerates TGF‐β2‐induced promotion in cell viability, migration, and invasion of lens epithelial cells via miR‐31/TXNIP signaling cascade
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DOI：
10.1002/jbt.23144
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发表时间：
2022-06
影响因子：
3.6
通讯作者：
Huajun Wang;Guangying Zheng;Miaomiao Sun;Y. Chi
中科院分区：
文献类型：
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作者：
Huajun Wang;Guangying Zheng;Miaomiao Sun;Y. Chi
后囊混浊是白内障手术后的主要并发症，可导致继发性视力下降。环状RNA（circRNA）在多种细胞生物学过程中起着重要的调控作用。circRNA最常见的工作机制是充当microRNA海绵。本文分析了circRNA RNA聚合酶III亚基A（POLR 3A）在PCO中的作用及机制。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑测定分析细胞活力。通过transwell和伤口愈合试验评估细胞运动性。进行双荧光素酶报告基因和RNA下拉测定，以验证microRNA-31（miR-31）与circ-POLR 3A或硫氧还蛋白相互作用蛋白（TXNIP）之间的相互作用。用转化生长因子-β2（TGF-β2）处理SRA 01/04细胞建立PCO细胞模型。我们发现TGF-β2增强了SRA 01/04细胞的活力、迁移和侵袭能力。在PCO组织和TGF-β2诱导的SRA 01/04细胞中，Circ-POLR 3A表达上调。TGF-β2主要通过上调circ-POLR 3A来促进SRA 01/04细胞的活力和运动性。Circ-POLR 3A通过直接与miR-31相互作用来负调节miR-31水平。Circ-POLR 3A缺失对TGF-β2诱导的SFA 01/04细胞的影响可通过沉默miR-31而部分逆转。miR-31与TXNIP的3′-非翻译区直接结合。TXNIP过表达在很大程度上减弱了TGF-β2诱导的SRA 01/04细胞中miR-31过表达介导的作用。Circ‐ POLR 3A可通过吸收miR-31而提高TXNIP的蛋白表达。外泌体参与介导在SRA 01/04细胞中circ‐ POLR 3A的递送。总之，circ-POLR 3A通过靶向miR-31/TXNIP轴，促进了TGF-β2诱导的SRA 01/04细胞的细胞活力、迁移和侵袭。
Posterior capsular opacification (PCO) is the major complication after cataract surgery and can result in secondary vision loss. Circular RNAs (circRNAs) are reported to play critical regulatory roles in multiple cell biological processes. The most common working mechanism of circRNAs is by acting as microRNA sponges. Here, we analyzed the role and mechanism of circRNA RNA polymerase III subunit A (POLR3A) in PCO. Cell viability was analyzed by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Cell motility was assessed by transwell and wound healing assays. Dual‐luciferase reporter and RNA‐pull‐down assays were performed to verify the interaction between microRNA‐31 (miR‐31) and circ‐POLR3A or thioredoxin interacting protein (TXNIP). PCO cell model was established by treating SRA01/04 cells with transforming growth factor‐β2 (TGF‐β2). We found that TGF‐β2 enhanced SRA01/04 cell viability, migration, and invasion abilities. Circ‐POLR3A expression was upregulated in PCO tissues and TGF‐β2‐induced SRA01/04 cells. TGF‐β2 promoted the viability and motility of SRA01/04 cells largely by upregulating circ‐POLR3A. Circ‐POLR3A negatively regulated the miR‐31 level by directly interacting with it. Circ‐POLR3A absence‐induced influences in TGF‐β2‐induced SRA01/04 cells were partly reversed by silencing miR‐31. miR‐31 is directly bound to the 3′‐untranslated region of TXNIP. TXNIP overexpression largely attenuated miR‐31 overexpression‐mediated effects in TGF‐β2‐induced SRA01/04 cells. Circ‐POLR3A could elevate the protein expression of TXNIP by sponging miR‐31. Exosomes were involved in mediating the delivery of circ‐POLR3A in SRA01/04 cells. In conclusion, circ‐POLR3A contributed to TGF‐β2‐induced promotion of cell viability, migration, and invasion of SRA01/04 cells by targeting miR‐31/TXNIP axis.