Department of Gastroenterology, Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
J Gastroenterol Hepatol. 2019 Oct;34(10):1860-1868. doi: 10.1111/jgh.14692. Epub 2019 Jun 28.
Secreted protein acidic and rich in cysteine (SPARC) is a matricellular glycoprotein that plays a significant role in tumor development. SPARC has been indicated that promotes tumorigenesis, metastasis, and poor prognosis in prostate cancer and lung cancer. Therefore, we sought to investigate the molecular mechanisms of SPARC in regulating hepatocellular carcinoma (HCC).
We used spheroids cultured in serum-free culture medium to obtain liver cancer stem cells. Flow cytometric analysis was performed to investigate percentage of CD133 cells in liver cancer cells. Real-time polymerase chain reaction and western blot were used to assess gene expression in cell lines. Transwell and wound healing assays were performed to indicate cell migration of HCC.
Secreted protein acidic and rich in cysteine was upregulated in spheres formation in HCC cells. Overexpression of SPARC enhanced the ability to form tumor spheres and increased CD133 and Oct4 expressions. Besides, SPARC promoted the migration and epithelial-mesenchymal transition in HCC cells. Importantly, SPARC overexpression stimulated the formation of subcutaneous tumors in nude mice.
Our findings suggest that SPARC overexpression promotes tumor growth, inducing epithelial-mesenchymal transition and acquisition of a stem cell phenotype. What is more, research elucidating the biological mechanisms of SPARC may be beneficial to liver cancer treatment.
富含半胱氨酸的酸性分泌蛋白(SPARC)是一种细胞外基质糖蛋白,在肿瘤的发生发展中发挥着重要作用。SPARC 已被证实可促进前列腺癌和肺癌的肿瘤发生、转移和预后不良。因此,我们试图研究 SPARC 在调控肝细胞癌(HCC)中的分子机制。
我们使用无血清培养培养基培养球体来获得肝癌干细胞。采用流式细胞术分析检测肝癌细胞中 CD133 细胞的百分比。实时聚合酶链反应和蛋白质印迹法用于检测细胞系中的基因表达。Transwell 和划痕愈合实验用于指示 HCC 细胞的迁移。
在 HCC 细胞的球体形成中,富含半胱氨酸的酸性分泌蛋白上调。SPARC 的过表达增强了形成肿瘤球体的能力,并增加了 CD133 和 Oct4 的表达。此外,SPARC 促进了 HCC 细胞的迁移和上皮-间充质转化。重要的是,SPARC 的过表达刺激了裸鼠皮下肿瘤的形成。
我们的研究结果表明,SPARC 的过表达促进肿瘤生长,诱导上皮-间充质转化和获得干细胞表型。更重要的是,研究 SPARC 的生物学机制可能有益于肝癌的治疗。