Wu Jie, Sun Lulu, Liu Tingting, Dong Gang
Department of Ultrasound Intervention, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, People's Republic of China.
Onco Targets Ther. 2021 Jan 11;14:221-237. doi: 10.2147/OTT.S269589. eCollection 2021.
Cancer cells could show the characteristics of cancer stem cells (CSCs) through epithelial-mesenchymal transition (EMT). EZH2 was associated with EMT. Ultrasound-targeted microbubble destruction (UTMD) could enhance gene transfection efficiency. Here, we explored the effect of UTMD-mediated shEZH2 on liver CSCs.
EZH2 expression in liver cancer and the overall survival of liver cancer patients were analyzed by bioinformatics. Liver CSCs (CD133HuH7) were sorted by flow cytometry. After transfection of shEZH2 through UTMD (UTMD-shEZH2) or liposome (LIP-shEZH2), the viability, proliferation, sphere formation, migration, and invasion of CD133HuH7 cells were detected by MTT, colony formation, tumor-sphere formation, wound healing, and transwell assays, respectively. A mice subcutaneous-xenotransplant tumor model was established by injecting CD133HuH7 or CD133HuH7 cells into the limbs of mice. Tumor weight and volume were documented. The expressions of EZH2, EMT-related factors, and STAT3/PI3K/AKT pathway-related factors in CD133HuH7 cells or tumor tissues were detected by RT-qPCR, Western blot, or immunohistochemical.
EZH2 was high-expressed in liver cancer, and the patients with high expression of EZH2 had a poor survival. CD133 HuH7 cells had higher EZH2 expression, higher viability, and stronger sphere-forming and tumor-forming abilities than CD133 HuH7 cells. ShEZH2 inhibited the viability, proliferation, sphere formation, migration, and invasion of CD133 HuH7 cells, decreased the weight and volume of the xenotransplant tumor, inhibited the expressions of EZH2, Vimentin, N-Cadherin, Twist-1, p-STAT3, p-PI3K, and p-AKT, and increased E-Cadherin expression. UTMD-shEZH2 caused a stronger effect on CD133 HuH7 cells than LIP-shEZH2.
UTMD-mediated shEZH2 inhibited the stemness and EMT of liver CSCs in vitro and in vivo through regulating the STAT3/PI3K/AKT pathway.
癌细胞可通过上皮-间质转化（EMT）表现出癌症干细胞（CSCs）的特征。EZH2与EMT相关。超声靶向微泡破坏（UTMD）可提高基因转染效率。在此，我们探讨了UTMD介导的shEZH2对肝脏CSCs的影响。
通过生物信息学分析肝癌中EZH2的表达及肝癌患者的总生存期。采用流式细胞术分选肝脏CSCs（CD133HuH7）。通过UTMD（UTMD-shEZH2）或脂质体（LIP-shEZH2）转染shEZH2后，分别采用MTT法、集落形成法、肿瘤球形成法、伤口愈合法和Transwell法检测CD133HuH7细胞的活力、增殖、球形成、迁移和侵袭能力。通过将CD133HuH7或CD133HuH7细胞注射到小鼠肢体建立小鼠皮下异种移植瘤模型。记录肿瘤重量和体积。通过RT-qPCR、Western blot或免疫组化检测CD133HuH7细胞或肿瘤组织中EZH2、EMT相关因子以及STAT3/PI3K/AKT通路相关因子的表达。
EZH2在肝癌中高表达，EZH2高表达的患者生存期较差。与CD133 HuH7细胞相比，CD133 HuH7细胞具有更高的EZH2表达、更高的活力以及更强的球形成和肿瘤形成能力。ShEZH2抑制了CD133 HuH7细胞的活力、增殖、球形成、迁移和侵袭，降低了异种移植瘤的重量和体积，抑制了EZH2、波形蛋白、N-钙黏蛋白、Twist-1、p-STAT3、p-PI3K和p-AKT的表达，并增加了E-钙黏蛋白的表达。UTMD-shEZH2对CD133 HuH7细胞的作用比LIP-shEZH2更强。
UTMD介导的shEZH2通过调节STAT3/PI3K/AKT通路在体外和体内抑制肝脏CSCs的干性和EMT。
