Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
Department of General Surgery, Affiliated Hospital 2 of Nantong University, Nantong, China.
J Cell Mol Med. 2020 Apr;24(8):4830-4838. doi: 10.1111/jcmm.15161. Epub 2020 Mar 11.
Nowadays, research on CSCs is still in an initial stage, and there are few studies reporting the successful isolation and identification of CSCs. In the present study, we attempted to isolate CSCs through cultivating the cell line MKN45 in defined serum-free medium and study the expression of stem cell markers or related proteins (Oct3/4, Sox2, Nanog and CD44) in CSCs. Moreover, immunofluorescence staining was performed to validate the stem cell markers of spheroid body-forming cells. Further experiments were used to evaluate the SPOP expression in tumorsphere cells. In addition, ADAMTS9-AS2 is a lncRNA that contributes to the genesis and development of many cancers, including gastric cancer (GC). We found ADAMTS9-AS2 functioned as an anti-oncogene and positively correlated with the expression of SPOP in GC tissues by combining bioinformatics analyses. Furthermore, we reported that ADAMTS9-AS2 regulated the expression of SPOP in GC cells and tumorsphere cells to inhibit GC progression. Together, our results demonstrated that SPOP and ADAMTS9-AS2 can be potential targets for GC treatment.
目前,关于 CSCs 的研究仍处于起步阶段,少有研究成功分离和鉴定 CSCs。本研究通过在无血清限定培养基中培养 MKN45 细胞系,尝试分离 CSCs,并研究 CSCs 中干细胞标志物或相关蛋白(Oct3/4、Sox2、Nanog 和 CD44)的表达。此外,我们通过免疫荧光染色来验证球体形成细胞的干细胞标志物。进一步的实验用于评估肿瘤球细胞中 SPOP 的表达。此外,ADAMTS9-AS2 是一种长链非编码 RNA,它有助于许多癌症的发生和发展,包括胃癌(GC)。通过结合生物信息学分析,我们发现 ADAMTS9-AS2 在 GC 组织中作为一种抑癌基因,与 SPOP 的表达呈正相关。此外,我们报告 ADAMTS9-AS2 调节 GC 细胞和肿瘤球细胞中 SPOP 的表达,以抑制 GC 的进展。综上所述,我们的结果表明 SPOP 和 ADAMTS9-AS2 可能成为 GC 治疗的潜在靶点。