外泌体lncRNA HOTAIR促进食管癌恶性转移的潜在分子机制研究  

Potential molecular mechanism of lncRNAs HOTAIR in malignant metastasis of esophageal cancer

作　　者：卢开进 申江峰[1] 韩光 陈泉[1] LU Kaijin;SHEN Jiangfeng;HAN Guang;CHEN Quan(Department of Thoracic Surgery,The Affiliated Taizhou People’s Hospital of Nanjing Medical University,Taizhou 225300,China)

机构地区：[1]泰州市人民医院(南京医科大学附属泰州人民医院)胸外科,江苏泰州225300

出　　处：《细胞与分子免疫学杂志》2025年第3期236-244,共9页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省卫生健康委科研课题(WJ2023J0105)。

摘　　要：目的 阐明食管癌相关成纤维细胞(CAF)来源外泌体(Exo)携带HOX转录本反义基因RNA(lncRNA HOTAIR)促进食管鳞状细胞癌(ESCC)转移的分子机制。方法 从肿瘤组织中收集CAF,和从距离肿瘤至少5 cm的邻近正常组织中获得非癌相关成纤维细胞(NF)。将从CAF或NF收集的条件培养基中分离Exo(CAF-Exo和NF-Exo),将CAF-Exo和NF-Exo与人ESCC细胞系TE-1细胞一起孵育24 h,CCK-8法检测细胞增殖能力,划痕实验、 Transwell^(TM)实验检测细胞的侵袭和迁移能力。将TE-1细胞分为以下2组:NC组和KD组。NC组和KD组分别转染对照siRNA和靶向HOTAIR的siRNA。测定HOTAIR敲低对细胞增殖、侵袭、迁移、糖酵解的影响。结果 CAF-Exo比NF-Exo更显著地促进TE-1细胞的增殖。与NF-Exo组相比,CAF-Exo处理的TE-1细胞的侵袭和迁移能力显著提高。此外,CAF-Exo处理抑制了上皮钙黏蛋白(E-cadherin)的表达并增强了神经钙黏蛋白(N-cadherin)的表达。HOTAIR在CAF中的表达显著高于NF中的表达。与NF-Exo相比,CAF-Exo中HOTAIR的表达水平也显著上调。与NC组相比,KD组TE-1细胞的增殖、侵袭和迁移能力显著降低。与NC组相比,KD组TE-1细胞中作为糖酵解的关键限速酶的己糖激酶2(HK2)、细胞外酸化率(ECAR)和三磷酸腺苷/二磷酸腺苷(ATP/ADP)比率显著下调。结论CAF来源Exo的HOTAIR可能通过调节ESCC细胞中糖酵解参与转移和上皮间质转化(EMT)。Objective To elucidate the molecular mechanism by which exosomes(Exo)derived from cancer-associated fibroblasts(CAF)carrying HOX transcript antisense intergenic RNA(lncRNA HOTAIR)promote the metastasis of esophageal squamous cell carcinoma(ESCC).Methods CAFs were collected from tumor tissues,and non-cancer associated fibroblasts(NFs)were obtained from adjacent normal tissues at least 5 cm away from the tumor.Exosomes(CAFs-Exo and NFs-Exo)were isolated from conditioned media collected from CAFs or NFs.CAFs-Exo and NFs-Exo were incubated with human ESCC cell line TE-1 for 24 hours,and CCK-8 was used to determine the cell proliferation ability.Scratch test and Transwell test were performed to determine the cell migration and invasion ability.TE-1 cells were divided into the following two groups:NC group and KD group.The NC group and KD group were transfected with control siRNAs or siRNAs targeting HOTAIR respectively.The effects of HOTAIR knock-down on cell proliferation,migration,invasion and glycolysis were determined.Results CAFs-Exo promoted the proliferation of TE-1 cells more significantly than NFs-Exo.Compared with NFs-Exo group,the migration and invasion ability of TE-1 cells treated with CAFs-Exo were improved significantly.In addition,CAFs-Exo treatment inhibited the expression of E-cadherin and enhanced the expression of N-cadherin.The expression of HOTAIR in CAFs was significantly higher than that in NFs.Compared with NFs-Exo,the expression level of HOTAIR in CAFs-Exo increased significantly. Compared with NC group, the proliferation, migration and invasion of TE-1 cells in KD group decreased significantly. Compared with NC group, hexokinase 2 (HK2), extracellular acidification rate (ECAR) and ATP/ADP ratio of TE-1 cells in KD group decreased significantly. Conclusion HOTAIR, an exosome derived from CAFs, may be involved in metastasis and EMT by regulating glycolysis in ESCC cells.

关 键 词：食管癌 食管癌相关成纤维细胞(CAF) lncRNA HOTAIR 糖酵解 

分 类 号：R392[医药卫生—免疫学] R735.1[医药卫生—基础医学]