河北医科大学学报 ›› 2024, Vol. 45 ›› Issue (6): 687-694.doi: 10.3969/j.issn.1007-3205.2024.06.011

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细胞外囊泡装载仑伐替尼对肝癌细胞株HepG2增殖、侵袭及凋亡的影响

  

  1. 江汉大学附属湖北省第三人民医院肿瘤科，湖北 武汉 430000

  • 出版日期:2024-06-25 发布日期:2024-06-25
  • 作者简介:孙建海（1973-），男，江苏宜兴人，江汉大学附属湖北省第三人民医院主任医师，医学硕士，从事肿瘤疾病诊治研究。
  • 基金资助:
    湖北省卫健委2019年科研项目（WJ2019F184）

Effects of extracellular vesicles loaded with lenvatinib on the proliferation, invasion and apoptosis of hepatocellular carcinoma cell line HepG2

  1. Department of Oncology, the Third People′s Hospital of Hubei Province Affiliated to Jianghan University, Hubei Province, Wuhan 430000, China

  • Online:2024-06-25 Published:2024-06-25

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摘要： 目的 探讨细胞外囊泡装载仑伐替尼对肝癌细胞株HepG2增殖、侵袭及凋亡的影响。
 方法 设肝癌细胞株HepG2组、细胞外囊泡组（extracellular vesicles，EVs混悬液5 mL、106个/ mL）、仑伐替尼组（仑伐替尼5 mL、30 mg/L）、仑伐替尼载药囊泡组（EVs混悬液5 mL、106个/mL+仑伐替尼5 mL、30 mg/L），以上各组细胞每孔设6个平行样，培养72 h。培养结束后，测定细胞增殖、侵袭、迁移、凋亡水平，应用RT-PCR法和蛋白印迹法测定各组细胞 miR-482、CYR61表达水平。
 结果 与肝癌细胞株HepG2组比较，细胞外囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、凋亡率、miR-482、CYR61 mRNA和蛋白水平差异无统计学意义（P＞0.05）。与肝癌细胞株HepG2组和细胞外囊泡组比较，仑伐替尼组、仑伐替尼载药囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、CYR61 mRNA和蛋白水平降低，凋亡率、miR-482升高（P＜0.05）。与仑伐替尼组比较，仑伐替尼载药囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、CYR61 mRNA和蛋白水平降低，凋亡率、miR-482升高（P＜0.05）。
 结论 细胞外囊泡装载仑伐替尼后能明显增强仑伐替尼对肝癌细胞株HepG2的杀伤力，增加其抑制增殖、侵袭与促凋亡作用；其机制可能与细胞外囊泡装载仑伐替尼能促进肝癌细胞株HepG2高表达miR-482，低表达CYR61有关。


关键词: 癌, 肝细胞, 细胞外囊泡, 仑伐替尼

Abstract: Objective To investigate the effect of extracellular vesicle loaded with lenvatinib on the proliferation, invasion and apoptosis of hepatocellular carcinoma cell line HepG2. 
Methods Liver cancer cell line HepG2 group, extracellular vesicles group (EVs suspension 5 mL, 106/mL), lenvatinib group (lenvatinib 5 mL, 30 mg/L), and lenvatinib-loaded vesicle group (EVs suspension 5 mL, 106/mL+lenvatinib 5 mL, 30 mg/L). Six parallel samples were set up in each well of the above groups and cultured for 72 h. Cell proliferation, invasion, migration and apoptosis were measured after the end of culture. The expression levels of miR-482 and CYR61 in each group were detected by real-time PCR (RT-PCR) and Western blot. 
Results Compared with HepG2 group, the OD value, survival rate, the number of monoclonal clones formed, the number of transmembrane cells, the migration distance, the apoptosis rate, the mRNA and protein levels of miR-482 and CYR61 in the extracellular vesicle group had no significant changes (P＞0.05). Compared with the HepG2 group and the extracellular vesicle group, OD value, survival rate, number of monoclonal clones formed, number of transmembrane cells, migration distance, CYR61 mRNA and protein levels were decreased in the lenvatinib group and the lenvatinib-loaded vesicle group, while apoptosis rate and miR-482 were increased (P＜0.05). Compared with the lenvatinib group, the OD value, survival rate, the number of monoclonal clones formed, the number of transmembrane cells, the migration distance, CYR61 mRNA and protein levels were decreased in the lenvatinib-loaded vesicle group, while the apoptosis rate and miR-482 were increased (P＜0.05). 
Conclusion Extracellular vesicles loaded with lenvatinib can significantly enhance the killing effect of chemotherapy lenvatinib on hepatocellular carcinoma cell line HepG2, and increase its inhibitory effect on proliferation, invasion and pro-apoptosis. The mechanism may be related to the increased expression of miR-482 and decreased expression of CYR61 in HepG2 cells loaded with lenvatinib in extracellular vesicles. 


Key words: carcinoma, hepatocellular, extracellular vesicles, lenvatinib

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