环状 RNA 细胞衰老相关基因 1 通过调控 CTNNBIP1 控制抗苯并[a]芘-7,8-反式-9,10-环氧化物转化 16HBE 细胞迁移。
The Circ_CARM1 controls cell migration by regulating CTNNBIP1 in anti-benzo[a]pyrene-trans-7,8-dihydrodiol-9,10-epoxide-transformed 16HBE cells.
机构信息
Institute for Chessmical Carcinogenesis, Guangzhou Medical University, Guangzhou, 511436, China.
Institute for Chessmical Carcinogenesis, Guangzhou Medical University, Guangzhou, 511436, China.
出版信息
Toxicol Lett. 2021 Sep 15;348:40-49. doi: 10.1016/j.toxlet.2021.05.007. Epub 2021 May 27.
BACKGROUND
Circular RNAs (circRNAs) have an important role in the development and progression of human tumors, including lung cancer. Yet, their role in lung cancer induced by benzo(a)pyrene (B[a]P) remains unclear. In this study, circRNA chips and qRT-PCR were used to examine downregulated circRNAs in malignantly transformed 16HBE cells (16HBE-T) induced by B[a]P. Five down-regulated circRNAs were found, among which hsa_circ_0004552 (circ_CARM1) had the most significant downregulation. Consequently, the role of circ_CARM1 on 16HBE-T cells biological behavior was further examined using several in vitro experiments.
MATERIALS AND METHODS
Detecting RNA expression via qRT-PCR. Fluorescence in situ hybridization (FISH) was used to identify the localization of circ_CARM1 in 16HBE-T. The effect of circ_CARM1 on cell behavior (cell migration, proliferation, and apoptosis) was explored by transfecting cells with a vector carrying an overexpression and then using wound healing, transwell migration assay, and flow cytometry. Also, the regulation mechanism for circ_CARM1, miR-1288-3p, and CTNNBIP1 was studied by Dual-Luciferase® Reporter (DLR™) Assay System and western blotting.
RESULTS
Reduced expression of circ_CARM1 is observed in 16HBE-T. The overexpression of circ_CARM1 further inhibited the migration of 16HBE-T cells but did not affect cell proliferation and apoptosis. Furthermore, bioinformatic analysis and Dual-Luciferase® Reporter (DLR™) Assay System showed that the competitive binding of circ_CARM1 and miR-1288-3p enhanced the expression of CTNNBIP1, thereby inhibiting the migration of 16HBE-T cells.
CONCLUSION
Downregulation of circ_CARM1 can stimulate the expression of miR-1288-3p, thereby reducing the expression of CTNNBIP1, spurring cell migration.
背景
环状 RNA（circRNAs）在包括肺癌在内的人类肿瘤的发生和发展中具有重要作用。然而，它们在苯并（a）芘（B[a]P）诱导的肺癌中的作用尚不清楚。在这项研究中，使用 circRNA 芯片和 qRT-PCR 检测了 B[a]P 诱导的恶性转化 16HBE 细胞（16HBE-T）中下调的 circRNAs。发现了 5 个下调的 circRNAs，其中 hsa_circ_0004552（circ_CARM1）下调最显著。因此，进一步通过几种体外实验研究了 circ_CARM1 对 16HBE-T 细胞生物学行为的作用。
材料和方法
通过 qRT-PCR 检测 RNA 表达。使用荧光原位杂交（FISH）鉴定 circ_CARM1 在 16HBE-T 中的定位。通过转染带有过表达载体的细胞，然后使用划痕愈合、Transwell 迁移测定和流式细胞术研究 circ_CARM1 对细胞行为（细胞迁移、增殖和凋亡）的影响。此外，通过双荧光素酶报告基因（DLR）检测系统和 Western blot 研究了 circ_CARM1、miR-1288-3p 和 CTNNBIP1 的调控机制。
结果
在 16HBE-T 中观察到 circ_CARM1 的表达降低。circ_CARM1 的过表达进一步抑制了 16HBE-T 细胞的迁移，但不影响细胞增殖和凋亡。此外，生物信息学分析和双荧光素酶报告基因（DLR）检测系统显示，circ_CARM1 和 miR-1288-3p 的竞争性结合增强了 CTNNBIP1 的表达，从而抑制了 16HBE-T 细胞的迁移。
结论
circ_CARM1 的下调可刺激 miR-1288-3p 的表达，从而降低 CTNNBIP1 的表达，促进细胞迁移。
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