环状RNA FAT1(hsa_circ_0001461)通过调控miR⁃181d⁃5p/miR⁃199a⁃5p促进口腔鳞状细胞癌进展
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R739.85

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浙江省医药卫生科技计划项目(2021KY297);宁波市医学重点扶植学科(2022-F20)


Circular RNA FAT1(hsa_circ_0001461)promotes the progression of oral squamous cell carcinoma by regulating miR⁃181d⁃5p/miR⁃199a⁃5p
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    摘要:

    目的:探讨环状RNA FAT1(hsa_circ_0001461)-miR-181d-5p/miR-199a-5p轴在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞生物学功能中的作用及机制。方法:收集 30 例 OSCC 患者的临床组织,采用实时定量 PCR 检测 circ- FAT1、miR-181d-5p及miR-199a-5p在OSCC组织和细胞中的表达。通过StarBase和双荧光素酶报告基因实验验证 miR-181d- 5p/miR-199a-5p与circFAT1的靶向关系。利用CCK-8、Transwell、流式细胞术检测 circFAT1-miR-181d-5p/miR-199a-5p轴对于 OSCC细胞活力、迁移、侵袭、凋亡和周期的影响。采用免疫印迹法结合LY294002处理检测PI3K/Akt/mTOR信号通路相关蛋白的表达。结果:在OSCC中circFAT1表达升高,miR-181d-5p、miR-199a-5p表达下降(P < 0.001)。circFAT1可以同时靶向miR- 181d-5p和miR-199a-5p(P < 0.01)。沉默circFAT1、过表达miR-181d及miR-199a-5p抑制了OSCC细胞活力、迁移、侵袭和周期进展、诱导凋亡,同时抑制PI3K/Akt/mTOR 通路的激活;过表达circFAT1、抑制miR-181d-5p及miR-199a-5p则发挥相反作用 (P < 0.05)。miR-181d-5p、miR-199a-5p可部分逆转circFAT1对OSCC细胞生物学功能的影响(P<0.05)。LY294002预处理逆转了过表达circFAT1对PI3K/Akt/mTOR通路和细胞活力的影响(P < 0.05)。结论:circFAT1通过调控miR-181d-5p和miR-199a-5p 促进OSCC细胞恶性进展。

    Abstract:

    Objective:To explore the role and mechanism of circular RNA FAT1(hsa_circ_0001461)-miR-181d-5p/miR-199a-5p axis in the biological functions of oral squamous cell carcinoma(OSCC)cells. Methods:The clinical tissues of 30 OSCC patients were collected,and the expressions of circFAT1,miR -181d -5p and miR -199a -5p in OSCC tissues and cells were detected by real -time quantitative PCR. starBase and dual luciferase reporter genes were used to verify the targeting relationship between miR-181d-5p/miR- 199a-5p and circFAT1. Functional experiments include CCK-8,Transwell,and flow cytometry to detect the effects of circFAT1-miR- 181d-5p/miR-199a-5p axis on OSCC cell viability,migration,invasion,apoptosis and cycle. Western blot combined with LY294002 treatment was used to detect the expression of PI3K/Akt/mTOR signaling pathway related proteins. Results:The circFAT1 expression increased,and miR-181d-5p and miR-199a-5p expressions decreased in OSCC(P < 0.001). CircFAT1 could target miR-181d-5p and miR-199a-5p(P < 0.01). Silencing circFAT1,miR-181d-5p mimic and miR-199a-5p mimic inhibited OSCC cell viability,migration, invasion and cycle progression,induced apoptosis,and inhibited the activation of PI3K/Akt/mTOR pathway;overexpressed circFAT1, miR-181d-5p inhibitor and miR-199a-5p inhibitor played the opposing roles(P < 0.05). MiR-181d-5p and miR-199a-5p partially reversed the effect of circFAT1 on the biological functions of OSCC cells(P < 0.05). LY294002 pretreatment reversed the effect of overexpression of circFAT1 on PI3K/Akt/mTOR pathway and cell viability(P < 0.05). Conclusion:circFAT1 promotes the malignant progression of OSCC cells by miR-181d-5p/miR-199a-5p.

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陈吉俊,王梁,马诞骅,高红燕,石宇远.环状RNA FAT1(hsa_circ_0001461)通过调控miR⁃181d⁃5p/miR⁃199a⁃5p促进口腔鳞状细胞癌进展[J].南京医科大学学报(自然科学版),2023,(2):196-206

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  • 在线发布日期: 2023-02-16
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