外泌体微小RNA-183-5p诱导淋巴管生成促进肺腺癌转移  

Exosome-transmitted microRNA-183-5p induces lung adenocarcinoma lymphangiogenesis and metastasis

作　　者：方乐平 沈泓妤 许菊青 徐伟 王琳 Fang Leping;Shen Hongyu;Xu Juqing;Xu Wei;Wang Lin(Department of Hematology and Oncology,Jiangsu Province Geriatric Hospital,Nanjing,210029,China;Gusu School,Nanjing Medical University,Suzhou,215026,China;Department of General Surgery,the First Affiliated Hospital of Nanjing Medical University,Nanjing,210029,China)

机构地区：[1]江苏省老年病医院血液肿瘤科,南京210029 [2]南京医科大学姑苏学院,苏州215026 [3]南京医科大学附属第一医院普外科,南京210029

出　　处：《中华实验外科杂志》2023年第10期2043-2046,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金项目(82103574);江苏省自然科学基金项目(BK20201076);南京医科大学科技发展基金项目(NMUB2018181)。

摘　　要：目的探讨外泌体微小RNA(miR)-183-5p在肺腺癌(LUAD)中作用。方法通过定量反转录聚合酶链反应(qRT-PCR)验证LUAD患者外周血miR-183-5p的表达,通过5-乙炔基-2’-脱氧尿苷(EdU)增殖、侵袭试验和淋巴管形成试验探讨其在LUAD中的作用。利用生物信息学预测和荧光素酶报告基因实验,确定miR-183-5p的靶基因。采用t检验或单因素方差分析进行统计分析。结果miR-183-5p在LUAD患者血浆表达高于健康组(2.746±1.111比1.346±0.474,t=8.195,P<0.05),且在淋巴结(LN)转移中表达高于LN阴性者(3.503±1.304比2.320±0.706,t=4.174,P<0.05),在LUAD血浆外泌体中表达也高于健康组(2.071±0.931比0.830±0.633,t=6.039,P<0.05)。EdU增殖实验表明miR-183-5p mimic及其对照组和miR-183-5p inhibitor及其对照组个数分别为83.000±4.000、55.670±5.508、25.330±5.508、50.330±5.132,差异有统计学意义(F=65.180,P<0.05),4组中淋巴管形成实验结果分别为1.400±0.141、0.983±0.080、0.733±0.103、1.006±0.030,差异有统计学意义(F=24.150,P<0.05),Transwell各组穿膜细胞数分别为84.000±6.557、53.000±4.000、40.670±6.429、59.000±7.000,差异有统计学意义(F=26.710,P<0.05)。C端小磷酸酶类似物(CTDSPL)是miR-183-5p的靶点,上调或下调人淋巴内皮细胞(HLEC)中miR-183-5p,CTDSPL的表达呈相反改变(1.010±0.076比0.383±0.025,1.540±0.098比0.983±0.071,F=127.300,P<0.05)。结论外泌体miR-183-5p在LUAD癌组织和血浆中均高表达,并可能通过靶向CTDSPL诱导淋巴管生成,可能成为LUAD的标志物。Objective To explore the role of exosomal microRNA(miR)-183-5p in lung adenocarcinoma(LUAD).Methods We verified miR-183-5p expression in peripheral plasma of LUAD patients by quantitative reverse-transcription polymerase chain reaction(qRT-PCR),EdU proliferation,migration assays,tube formation were performed to explore the role of exosomal miR-183-5p.Bioinformatic prediction and luciferase reporter assay were used to identify target gene.Statistical analysis was performed by t test or one-way ANOVA.Results The expression of miR-183-5p in plasma of LUAD patients was higher than that of healthy group(2.746±1.111 vs.1.346±0.474,t=8.195,P<0.05),also higher in patients with lymph node(LN)metastasis than LN negative patients(3.503±1.304 vs.2.320±0.706,t=4.174,P<0.05).Besides,the expression in LUAD plasma exosomes was higher than that of healthy group(2.071±0.931 vs.0.830±0.633,t=6.039,P<0.05).The number of EdU-positive cells in miR-183-5p mimic,negative control(NC)mimic,miR-183-5p inhibitor,NC inhibitor groups were 83.000±4.000,55.670±5.508,25.330±5.508,50.330±5.132,respectively.The difference was statistically significant(F=65.180,P<0.05).The results of lymphatic vessel formation in the 4 groups were 1.400±0.141,0.983±0.080,0.733±0.103,1.006±0.030,respectively,and the differences were statistically significant(F=24.150,P<0.05).The number of Transwell penetrating cells in each group was 84.000±6.557,53.000±4.000,40.670±6.429,59.000±7.000,respectively,and the difference was statistically significant(F=26.710,P<0.05).C-terminal domain small phosphatase-like(CTDSPL)is the target of miR-183-5p.Up-regulated or down-regulated miR-183-5p in human lymphatic endothelial cells(HLEC),the expression of CTDSPL was reversed(1.010±0.076 vs.0.383±0.025,1.540±0.098 vs.0.983±0.071,F=127.300,P<0.05).Conclusion Exosomal miR-183-5p is upregulated both in LUAD tumor tissue and plasma,and might induce lymphangiogenesis by targeting CTDSPL,it may become a good marker for LUAD.

关 键 词：外泌体 肺腺癌 淋巴管形成 微小RNA-183-5p 

分 类 号：R734.2[医药卫生—肿瘤]