肿瘤外泌体来源的miR-126-5p通过抑制NLRC3促进口腔鳞癌细胞的增殖和迁移

Tumor exosome-derived miR-126-5p promotes proliferation and migration of oral squamous carcinoma cells by inhibiting NLRC3

目的探索口腔鳞癌外泌体中miR-126-5p对肿瘤细胞生物学功能的影响。方法qPCR分析口腔鳞癌组织芯片、临床样本及口腔鳞癌外泌体中miR-126-5p的表达情况;荧光素酶报告基因法确定与miR-126-5p相互作用的分子,qPCR检测外泌体与共孵育后的肿瘤细胞中NOD样受体家族蛋白3(NLRC3)的表达情况;通过体外细胞增殖及划痕实验,以及体内肿瘤生长反映含有miR-126-5p外泌体对肿瘤细胞生物学功能的影响。结果芯片分析表明口腔鳞癌组织中miR-126-5p高表达,患者组织(P<0.05)及口腔鳞癌细胞系中miR-126-5p同样高表达(P<0.001),荧光素酶报告实验表明miR-126-5p与NLRC3相互作用,且miR-126-5p对NLRC3存在负性调控关系(P<0.01);此外,含有miR-126-5p的外泌体与细胞共孵育或沉默NLRC3可以促进肿瘤细胞增殖(P<0.001)及迁移(P<0.0001),且含有miR-126-5p的外泌体可以抑制NLRC3的体内表达(P<0.001),促进肿瘤的生长(P<0.001)。结论肿瘤细胞分泌的外泌体miR-126-5p可以抑制NLRC3的表达从而促进口腔鳞癌的进展。

Objective To explore the effect of miR-126-5p in exosome derived from oral squamous cell carcinoma(OSCC)on the biological function of tumor cells.Methods qPCR was used to analyze the expression levels of miR-126-5p in tissue microarray,clinical samples and exosome of oral squamous cell carcinoma.The molecules interacting with miR-126-5p were identified by luciferase reporter gene method,and the expression of NLRC3 in exosome and tumor cells after co-incubation was detected by qPCR.The effect of miR-126-5p exosome on the biological function of tumor cells was detected by cell proliferation and migration test in vitro,scratch test and tumor growth in vivo.Results The microarray analysis showed that miR-126-5p was highly expressed in OSCC tissue,and miR-126-5p was also highly expressed in the patients′tissue(P<0.05)and OSCC cell lines(P<0.001).The luciferase assay showed that miR-126-5p interacted with NLRC3,moreover miR-126-5p had the negative regulation relation on NLRC3(P<0.01).In addition,exosomes containing miR-126-5p coincubation with cells or silencing NLRC3 could promote the proliferation(P<0.001)and migration(P<0.0001)of tumor cells.Moreover the exosomes containing miR-126-5p could inhibit the in vivo expression of NLRC3(P<0.001)and promote the tumor growth(P<0.001).Conclusion Exosomes miR-126-5p secreted by tumor cells can inhibit the expression of NLRC3,thus promote the progression of OSCC.