ABSTRACT

Hepatocellular carcinoma (HCC) is a malignancy of considerable concern due to its continuous increase in morbidity and mortality. This study attempts to identify the molecules that play a key role in the progression of HCC, explore its potential mechanism, and provide more target choices for targeted therapy. Using overexpression plasmid and shRNA, CKS1B was respectively overexpressed and knocked down to explore its biological function roles in HCC progression and development. MTT and colony formation assays showed that knockdown of CKS1B inhibited the survival and proliferation of HCC cell lines (Hep3B and Huh7). The flow cytometry and western blot analysis showed that knockdown of CKS1B significantly induced the apoptosis of Hep3B and Huh7 cells. The wound healing and transwell invasion assays showed that knockdown of CKS1B had a significant inhibitory effect on the migration and invasion of Hep3B and Huh7 cells. These functional tests confirmed that CKS1B acts as an oncogene that regulates the malignant progression of HCC. Moreover, this study also demonstrated that knockdown of CKS1B inhibited the activation of JAK/STAT3 pathway, evidenced by the significantly downregulated p-STAT3 protein expression. Furthermore, knockdown of CKS1B also downregulated STAT3 target genes TIMP-1, Bcl-2 and VEGF, which were involved in controlling cell apoptosis and migration. On the contrary, overexpression of CKS1B caused the completely opposite results. Taken together, CKS1B acts as an oncogene to promote the proliferation and metastasis of HCC cells by activating JAK/STAT3 signaling pathway.

摘要

由于其发病率和死亡率的持续增加，肝细胞癌 (HCC) 是一种备受关注的恶性肿瘤。本研究试图找出在HCC进展中起关键作用的分子，探索其潜在机制，为靶向治疗提供更多的靶点选择。使用过表达质粒和shRNA，分别过表达和敲低CKS1B，以探索其在HCC进展和发展中的生物学功能作用。MTT 和集落形成分析表明，CKS1B 的敲低抑制了 HCC 细胞系（Hep3B 和 Huh7）的存活和增殖。流式细胞术和蛋白质印迹分析表明，CKS1B 的敲低显着诱导了 Hep3B 和 Huh7 细胞的凋亡。伤口愈合和 transwell 侵袭实验表明，CKS1B 的敲低对 Hep3B 和 Huh7 细胞的迁移和侵袭具有显着的抑制作用。这些功能测试证实了 CKS1B 作为调节 HCC 恶性进展的癌基因。此外，这项研究还表明，CKS1B 的敲低抑制了 JAK/STAT3 通路的激活，这可以通过 p-STAT3 蛋白表达的显着下调来证明。此外，CKS1B 的敲低还下调了 STAT3 靶基因 TIMP-1、Bcl-2 和 VEGF，这些基因参与控制细胞凋亡和迁移。相反，CKS1B 的过度表达导致完全相反的结果。综上所述，CKS1B 作为致癌基因通过激活 JAK/STAT3 信号通路促进 HCC 细胞的增殖和转移。