Hu Yilin, Zang Weijie, Feng Ying, Mao Qinsheng, Chen Junjie, Zhu Yizhun, Xue Wanjiang
Department of Gastrointestinal Surgery, Affliated Hospital of Nantong University, Medical School of Nantong University, 20 Xisi Street, Nantong, Jiangsu, 226001, China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, 226001, China.
Cancer Cell Int. 2024 May 27;24(1):184. doi: 10.1186/s12935-024-03359-5.
Cancer-induced pre-metastatic niches (PMNs) play a decisive role in promoting metastasis by facilitating angiogenesis in distant sites. Evidence accumulates suggesting that microRNAs (miRNAs) exert significant influence on angiogenesis during PMN formation, yet their specific roles and regulatory mechanisms in gastric cancer (GC) remain underexplored.
miR-605-3p was identified through miRNA-seq and validated by qRT-PCR. Its correlation with the clinicopathological characteristics and prognosis was analyzed in GC. Functional assays were performed to examine angiogenesis both in vitro and in vivo. The related molecular mechanisms were elucidated using RNA-seq, immunofluorescence, transmission electron microscopy, nanoparticle tracking analysis, enzyme-linked immunosorbent assay, luciferase reporter assays and bioinformatics analysis.
miR-605-3p was screened as a candidate miRNA that may regulate angiogenesis in GC. Low expression of miR-605-3p is associated with shorter overall survival and disease-free survival in GC. miR-605-3p-mediated GC-secreted exosomes regulate angiogenesis by regulating exosomal nitric oxide synthase 3 (NOS3) derived from GC cells. Mechanistically, miR-605-3p reduced the secretion of exosomes by inhibiting vesicle-associated membrane protein 3 (VAMP3) expression and affects the transport of multivesicular bodies to the GC cell membrane. At the same time, miR-605-3p reduces NOS3 levels in exosomes by inhibiting the expression of intracellular NOS3. Upon uptake of GC cell-derived exosomal NOS3, human umbilical vein endothelial cells exhibited increased nitric oxide levels, which induced angiogenesis, established liver PMN and ultimately promoted the occurrence of liver metastasis. Furthermore, a high level of plasma exosomal NOS3 was clinically associated with metastasis in GC patients.
miR-605-3p may play a pivotal role in regulating VAMP3-mediated secretion of exosomal NOS3, thereby affecting the formation of GC PMN and thus inhibiting GC metastasis.
癌症诱导的前转移微环境（PMN）通过促进远处部位的血管生成在促进转移中起决定性作用。越来越多的证据表明，微小RNA（miRNA）在PMN形成过程中对血管生成有显著影响，但其在胃癌（GC）中的具体作用和调控机制仍未得到充分研究。
通过miRNA测序鉴定出miR-605-3p，并通过qRT-PCR进行验证。分析其在GC中与临床病理特征及预后的相关性。进行功能实验以检测体内外血管生成情况。利用RNA测序、免疫荧光、透射电子显微镜、纳米颗粒跟踪分析、酶联免疫吸附测定、荧光素酶报告基因测定和生物信息学分析阐明相关分子机制。
miR-605-3p被筛选为可能调控GC血管生成的候选miRNA。miR-605-3p低表达与GC患者较短的总生存期和无病生存期相关。miR-605-3p介导的GC分泌外泌体通过调节源自GC细胞的外泌体一氧化氮合酶3（NOS3）来调控血管生成。机制上，miR-605-3p通过抑制囊泡相关膜蛋白3（VAMP3）的表达减少外泌体分泌，并影响多囊泡体向GC细胞膜的转运。同时，miR-605-3p通过抑制细胞内NOS3的表达降低外泌体中NOS3水平。人脐静脉内皮细胞摄取GC细胞来源的外泌体NOS3后，一氧化氮水平升高，诱导血管生成，建立肝脏PMN并最终促进肝转移的发生。此外，GC患者血浆中外泌体NOS3水平高与转移在临床上相关。
miR-605-3p可能在调控VAMP3介导的外泌体NOS3分泌中起关键作用，从而影响GC PMN的形成，进而抑制GC转移。
