Chen Lanlan, Huang Shenhui, Huang Jincheng, Chen Qiujuan, Zhuang Qihong
Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Xiamen University, No. 55 Zhenhai Road, Xiamen City, Fujian Province 361001, China.
ACS Omega. 2021 Jun 29;6(27):17217-17227. doi: 10.1021/acsomega.1c00905. eCollection 2021 Jul 13.
HOX transcript antisense RNA (HOTAIR) is a long noncoding RNA (lncRNA) that promotes tumor growth and metastasis. Exosomes can mediate intracellular communication in cancer by transferring active molecules. However, the role and mechanism of HOTAIR in nonsmall cell lung cancer (NSCLC) are still unclear. This study mainly explores the role and mechanism of exosome-derived HOTAIR in NSCLC.
after the material characterization of the CD63 immune lipid magnetic bead (CD63-IMB), the exosomes in serum of NSCLC patients were captured through CD63-IMB for the corresponding biological characterization. Real-time quantitative reverse transcription PCR (qRT-PCR) was performed to detect the expression level of HOTAIR in tumor tissues, serum, and serum exosome from NSCLC patients. Subsequently, exosome secreted by NCI-H1975 cells with highly expressed HOTAIR was selected to treat low-expression A549 cells and HOTAIR knockdown on NCI-H1975 cells. In this way, action mechanisms of HOTAIR can be investigated by means of qRT-PCR, colony formation assays, and flow cytometry.
exosomes can be isolated by CD63-IMB, and taken up by cells effectively; the qRT-PCR results demonstrate that HOTAIR expressions are significantly upregulated in tumor tissues, serums, and exosomes isolated from serums of NSCLC patients. Clinicopathological correlation analysis shows that the upregulation of HOTAIR is closely associated with lymphatic metastasis and tumor node metastasis (TNM) staging ( < 0.05). HOTAIR expressions show a significant increase in A549 cells treated with exosomes derived from NCI-H1975 cells, signifying that both proliferation and migration of A549 cells are promoted, and HOTAIR depletion could inhibit the proliferation and migration of lung cancer cells.
HOTAIR is highly expressed in tumor tissues, serums, and serum exosomes of NSCLC patients and its expression has a significant correlation with lymphatic metastasis and TNM staging. Moreover, the exosome may promote NSCLC proliferation and migration through HOTAIR transportation. Therefore, exosome-derived HOTAIR is expected to be a new molecular marker for NSCLC diagnosis, and exosomal transmission of HOTAIR may provide a new approach to NSCLC diagnosis.
HOX转录反义RNA（HOTAIR）是一种促进肿瘤生长和转移的长链非编码RNA（lncRNA）。外泌体可通过转运活性分子介导癌症中的细胞间通讯。然而，HOTAIR在非小细胞肺癌（NSCLC）中的作用及机制仍不清楚。本研究主要探讨外泌体来源的HOTAIR在NSCLC中的作用及机制。
对CD63免疫脂质磁珠（CD63-IMB）进行材料表征后，通过CD63-IMB捕获NSCLC患者血清中的外泌体进行相应生物学表征。采用实时定量逆转录PCR（qRT-PCR）检测NSCLC患者肿瘤组织、血清及血清外泌体中HOTAIR的表达水平。随后，选择高表达HOTAIR的NCI-H1975细胞分泌的外泌体处理低表达的A549细胞，并对NCI-H1975细胞进行HOTAIR敲低。通过qRT-PCR、集落形成实验和流式细胞术研究HOTAIR的作用机制。
CD63-IMB可分离出外泌体，且能被细胞有效摄取；qRT-PCR结果显示，NSCLC患者肿瘤组织、血清及血清分离出的外泌体中HOTAIR表达均显著上调。临床病理相关性分析表明，HOTAIR上调与淋巴转移和肿瘤淋巴结转移（TNM）分期密切相关（<0.05）。用NCI-H1975细胞来源的外泌体处理的A549细胞中HOTAIR表达显著增加，表明A549细胞的增殖和迁移均得到促进，而HOTAIR缺失可抑制肺癌细胞的增殖和迁移。
HOTAIR在NSCLC患者的肿瘤组织、血清及血清外泌体中高表达，其表达与淋巴转移和TNM分期显著相关。此外，外泌体可能通过转运HOTAIR促进NSCLC的增殖和迁移。因此，外泌体来源的HOTAIR有望成为NSCLC诊断的新分子标志物，HOTAIR的外泌体传递可能为NSCLC诊断提供新途径。
