Huang Xiao-Juan, Wang Yan, Wang Hui-Ting, Liang Zhao-Feng, Ji Cheng, Li Xiao-Xi, Zhang Lei-Lei, Ji Run-Bi, Xu Wen-Rong, Jin Jian-Hua, Qian Hui
Wujin Institute of Molecular Diagnostics and Precision Cancer Medicine of Jiangsu University, Wujin Hospital Affiliated with Jiangsu University, 2 Yong Ning North Road, Chang Zhou, 213017, Jiangsu, China.
Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, School of Medicine, Jiangsu University, 301 Xuefu Road, Zhenjiang, 212013, Jiangsu, China.
Cancer Cell Int. 2023 Aug 1;23(1):151. doi: 10.1186/s12935-023-02976-w.
Exosome, a component of liquid biopsy, loaded protein, DNA, RNA and lipid gradually emerges as biomarker in tumors. However, exosomal circRNAs as biomarker and function mechanism in gastric cancer (GC) are not well understood.
Differentially expressed circRNAs in GC and healthy people were screened by database. The identification of hsa_circ_000200 was verified by RNase R and sequencing, and the expression of hsa_circ_000200 was evaluated using qRT-PCR. The biological function of hsa_circ_000200 in GC was verified in vitro. Western blot, RIP, RNA fluorescence in situ hybridization, and double luciferase assay were utilized to explore the potential mechanism of hsa_circ_000200.
Hsa_circ_000200 up-regulated in GC tissue, serum and serum exosomes. Hsa_circ_000200 in serum exosomes showed better diagnostic ability than that of tissues and serum. Combined with clinicopathological parameters, its level was related to invasion depth, TNM staging, and distal metastasis. Functionally, knockdown of hsa_circ_000200 inhibited GC cells proliferation, migration and invasion in vitro, while its overexpression played the opposite role. Importantly, exosomes with up-regulated hsa_circ_000200 promoted the proliferation and migration of co-cultured GC cells. Mechanistically, hsa_circ_000200 acted as a "ceRNA" for miR-4659a/b-3p to increase HBEGF and TGF-β/Smad expression, then promoted the development of GC.
Our findings suggest that hsa_circ_000200 promotes the progression of GC through hsa_circ_000200/miR-4659a/b-3p/HBEGF axis and affecting the expression of TGF-β/Smad. Serum exosomal hsa_circ_000200 may serve as a potential biomarker for GC.
外泌体作为液体活检的一个组成部分，携带蛋白质、DNA、RNA和脂质，逐渐成为肿瘤中的生物标志物。然而，外泌体环状RNA在胃癌（GC）中的生物标志物作用及功能机制尚未完全明确。
通过数据库筛选GC患者和健康人中差异表达的环状RNA。采用RNase R和测序验证hsa_circ_000200的鉴定结果，并用qRT-PCR评估hsa_circ_000200的表达。在体外验证hsa_circ_000200在GC中的生物学功能。利用蛋白质免疫印迹法、RNA免疫沉淀、RNA荧光原位杂交和双荧光素酶报告基因检测来探究hsa_circ_000200的潜在作用机制。
hsa_circ_000200在GC组织、血清和血清外泌体中表达上调。血清外泌体中的hsa_circ_000200比组织和血清具有更好的诊断能力。结合临床病理参数，其水平与浸润深度、TNM分期和远处转移相关。在功能上，敲低hsa_circ_000200可抑制GC细胞在体外的增殖、迁移和侵袭，而其过表达则起相反作用。重要的是，hsa_circ_000200上调的外泌体促进了共培养GC细胞的增殖和迁移。机制上，hsa_circ_000200作为miR-4659a/b-3p的“竞争性内源RNA”，增加肝素结合表皮生长因子（HBEGF）和转化生长因子-β/信号转导和转录激活因子（TGF-β/Smad）的表达，进而促进GC的发展。
我们的研究结果表明，hsa_circ_000200通过hsa_circ_000200/miR-4659a/b-3p/HBEGF轴并影响TGF-β/Smad的表达促进GC进展。血清外泌体hsa_circ_000200可能作为GC的潜在生物标志物。
