miR-513b-5p靶向趋化因子CXCL8抑制喉鳞癌细胞增殖、迁移和侵袭

MiR-513b-5p Inhibits Proliferation,Migration and Invasion of Laryngeal Squamous Cell Carcinoma by Targeting CXCL8

侵袭转移是喉鳞癌预后不良的主要危险因素,肿瘤微环境中的趋化因子与侵袭转移密切相关。CXCL8是一种细胞因子样的分泌蛋白质,在多种肿瘤的恶性发展过程中发挥重要作用,但其在喉鳞癌的作用尚未阐明。本文以喉鳞癌细胞为研究对象,阐明CXCL8在喉鳞癌细胞的作用,并寻找靶向CXCL8的miRNA,为喉鳞癌的诊断和治疗提供新靶点。GEPIA网站显示,CXCL8在头颈肿瘤组织中高表达(P<0.05)。实时荧光定量PCR发现,CXCL8在喉鳞癌细胞中高表达,酶联免疫检测也发现,CXCL8在喉鳞癌细胞上清中分泌量较高(P<0.001)。CCK8检测证实,降低CXCL8表达会明显抑制喉鳞癌细胞FD-LSC-1和AMC-HN8增殖(平均抑制率分别为34.0%,19.5%);EdU实验也证实,降低CXCL8表达明显抑制喉鳞癌细胞的增殖(P<0.05)。Transwell小室实验证实,降低CXCL8表达明显抑制喉鳞癌细胞FD-LSC-1迁移和侵袭(平均抑制率分别为40.0%,38.5%);降低CXCL8表达也明显抑制喉鳞癌细胞AMC-HN8迁移和侵袭(平均抑制率分别为37.5%,53.5%)。生物信息学网站预测,CXCL 8可能是miR-513b-5p靶基因,双荧光素酶报告检测证实,miR-513b-5p能够与CXCL 8-3′UTR结合,在喉鳞癌细胞中过表达miR-513b-5p,CXCL 8 mRNA表达量下降60%(P<0.01)。细胞功能恢复实验证实,miR-513b-5p削弱喉鳞癌细胞增殖、迁移和侵袭的能力可被外源表达的CXCL8有效逆转(P<0.05)。综上所述,miR-513b-5p通过靶向CXCL 8抑制喉鳞癌细胞增殖、迁移和侵袭。

Metastasis is the main risk factor for poor prognosis of laryngeal squamous cell carcinoma(LSCC).Chemokines are closely related to metastasis in the tumor microenvironment.CXCL8 is a cytokine-like secreted protein that plays key roles in the malignant development of a variety of tumors,but has not been elucidated in LSCC.In this paper,we elucidated the role of CXCL8 in LSCC cells and found miRNAs that targeted CXCL8,which may become new targets for the diagnosis and treatment of LSCC.Firstly,the GEPIA showed that CXCL8 was highly expressed in head and neck cancer(P<0.05).The real-time fluorescence quantification(qRT-PCR)found that CXCL 8 was highly expressed in LSCC cells.The enzyme-linked immunoassay also found that CXCL8 was highly secreted in the supernatant of LSCC cells(P<0.001).Then,the CCK8 assay confirmed that knockdown of CXCL 8 significantly inhibited the proliferation of FD-LSC-1 and AMC-HN8 cells(the average inhibition rate was 34.0%and 19.5%,respectively);The EdU assay also confirmed that knockdown of CXCL 8 significantly inhibited the proliferation of LSCC cells(P<0.05).The transwell assay confirmed that knockdown of CXCL 8 also significantly inhibited the migration and invasion of FD-LSC-1 cell(average inhibition rate was 40.0%,38.5%,respectively);Knockdown of CXCL 8 also significantly inhibited the migration and invasion of AMC-HN8 cell(average inhibition rate was 37.5%,53.5%,respectively).The analysis of bioinformatics predicted that CXCL 8 may be a target of miR-513b-5p.The dual luciferase reporter assay confirmed that miR-513b-5p could bind to the CXCL 8-3′UTR.QRT-PCR assay also confirmed that overexpression of miR-513b-5p could decrease the 60%of the CXCL 8 expression(P<0.01).Cell function rescue assays found that overexpressed of CXCL 8 could effectively reversed proliferation,migration and invasion of LSCC cells weakened by miR-513b-5p(P<0.05).In summary,miR-513b-5p inhibited the proliferation,migration and invasion of LSCC cells by targeting CXCL 8.