miR-107靶向FoxM1对鼻咽癌细胞生长和侵袭及迁移影响

目的microRNA与鼻咽癌(nasopharyngeal carcinoma,NPC)发生发展密切相关,但miR-107在鼻咽癌中的作用机制仍不清楚。本研究检测miR-107在鼻咽癌组织中的表达水平,并初步探讨其对鼻咽癌细胞增殖、迁移和侵袭影响的分子机制。方法收集2013-02-02-2013-12-24于佛山市第一人民医院经病理活检确诊的86例NPC组织和42例慢性鼻咽炎组织。采用原位杂交技术检测组织中miR-107的表达。分析miR-107表达与鼻咽癌患者临床特征及5年生存率的关系。双荧光素酶实验检测miR-107和人叉头框蛋白M1(forkhead box M1,FoxM1)在鼻咽癌细胞中的相互作用。在鼻咽癌细胞系CNE-2中转染miR-107 mimics和mimics NC,并设立mock对照组,采用实时荧光定量PCR法检测FoxM1 mRNA的表达量,蛋白质印迹实验检测FoxM1蛋白的表达量。然后通过CCK8、划痕和Transwell小室法检测鼻咽癌细胞的增殖、迁移和侵袭能力。结果miR-107在鼻咽癌组织中的阳性表达率(15.12%,13/86)低于慢性鼻咽炎组织(83.33%,35/42),差异有统计学意义,χ^2=96.37,P<0.001;与T分期(χ^2=24.816,P<0.001)和N分期(χ^2=29.368,P<0.001)有关联,且鼻咽癌miR-107阳性患者的5年生存率高于miR-107阴性组,χ^2=6.380,P=0.043。miR-107靶向调控FoxM1的3′UTR,降低了荧光素酶的表达活性,F=363.600,P<0.001。miR-107 mimics组FoxM1mRNA(F=267.600,P<0.001)和蛋白表达水平(F=101.300,P<0.001)低于mimics NC和mock组。与mimics NC和mock组相比,miR-107 mimics组细胞的增殖(F=1192.000,P<0.001)和迁移、侵袭(F=210.600,P<0.001)能力均降低。结论miR-107在鼻咽癌患者中低表达,通过靶向抑制FoxM1的表达而抑制鼻咽癌细胞的增殖、迁移和侵袭能力。

miR-107 affects the proliferation,invasion and migration of human nasopharyngeal carcinoma cells by targeting FoxM1

OBJECTIVE MicroRNA is closely related to nasopharyngeal carcinoma(NPC)development,and the role of miR-107 in NPC is still unclear.This study aimed to investigate the expression of miR-107 in human NPC and its effect on the proliferation,migration and invasion.METHODS Eighty-six NPC patients and forty-two non-NPC patients underwent surgical treatment at the First Ppeople’s Hospital of Foshan City from February 2 th,2013 to December 24 th,2013 were selected.In situ hybridization was used to detected the expression of miR-107 in NPC patients tissues and Non-NPC patients tissues was detected by in situ hybridization,and the relationship between miR-107 and the clinical pathological parameters in NPC was analyzed.Dual luciferase reporter gene assay was used to verify the interaction between miR-107 and forkhead box protein M1(FoxM1).miR-107 mimics and unrelated sequence control(mimics NC)were transfected into CNE-2 cells using LipofectamineTM2000.The expression of FoxM1 were detected using real-time PCR and western blot analysis.The proliferation of the NPC cells was determined by cell counting and colony formation assay;wound healing assay and Transwell assay were used to analyze the changes in the cell migration and invasion abilities in each group.RESULTS The expression of miR-107 in NPC tissues(15.12%,13/86)was lower than that in Non-NPC tissues(83.33%,35/42;χ^2=96.37,P<0.001).Tumor tissue miR 107 expression was significantly associated with T stages(χ^2=24.816,P<0.001)and N stages(χ^2=29.368,P<0.001).Patients without miR-107 expression in tumours had significantly higher 5-year survival rate than those with miR-107 expression(χ^2=6.380,P=0.043).The miR-107 regulated the 3′UTR of FoxM1 and reduced the expression activity of luciferase(F=363.600,P<0.001).Compared with mimics NC and mock cells,the NPC cell lines with miR-107 mimics showed significantly lowered FoxM1 expressions at both the mRNA(F=267.600,P<0.001)and(F=101.300,P<0.001)protein levels.In addition,miR-107 mimics cells proliferation(F=1192.000,P<0.001),invasion and migration(F=210.600,P<0.001)were significantly decreased.CONCLUSION miR-107 is broadly distributed in NPC patients tissues at a relatively low level,and inhibit the proliferation,invasion and migration of NPC cells by targeting FoxM1.