Tang Zuxiong, He Jun, Zou Jiayue, Yu Shufei, Sun Xiaoming, Qin Lei
Department of Hepato-Pancreatico-Biliary Surgery, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Department of General Surgery, Suzhou Wuzhong People's Hospital, Suzhou, Jiangsu, China.
PeerJ. 2021 Apr 13;9:e11200. doi: 10.7717/peerj.11200. eCollection 2021.
Cancer cell resistance to chemotherapy drugs such as Gemcitabine, Oxaliplatin, Cisplatin, Doxorubicin, and 5-fluorouracil account for the main reason of chemotherapy failure for HCC patients, especially for those with advanced HCC or metastasis patients. This emerging resistance limits the effectiveness and clinical application of these chemotherapy drugs. Previous studies reported that drug-resistant tumor cell-derived exosomes could transfer their resistance property to tumor sensitive cells in some cancer, including lung and gastric cancer. This study sought to explore whether HepG2/DDP cell-derived exosomes transmit cisplatin (DDP) resistance to HepG2 and other HCC sensitive cells, and provide considerable guidance for HCC nursing with Cisplatin DDP clinically.
The HepG2 DDP-resistant cell line (HepG2/DDP) was established, and the exosomes from both HepG2/DDP and HepG2 cells were isolated and named ES-2, ES-1, respectively. HepG2 or SMMC-7721 or Huh7 cells were treated with DDP or DDP + ES-2, and HepG2/DDP cells were treated with ES-1. Then, the activation of drug resistance via HepG2/DDP exosomes transfer to HepG2, SMMC-7721 and Huh7 cells were assessed by cell viability assay and ROS formation. Moreover, the relative expression of P-glycoprotein (P-gp) was measured by western blot analysis.
HepG2/DDP cell-derived exosomes were successfully isolated from cisplatin-resistant HepG2 cells, and named ES-2. Cell viability of HepG2 or SMMC-7721 or Huh7 cells treated with DDP + ES-2 was enhanced compared with that of DDP treatment group. Also, the concentration of ROS generated in cells under DDP or DDP + ES-2 treatment was strongly increased compared with that of control, although the concentration of ROS was clearly smaller in DDP + ES-2 treatment group compared with DDP treatment. At the same time, the expression of P-gp was upregulated on the ES-2 surface.
The results mentioned above clarified that HepG2/DDP cell-derived exosomes conferred cisplatin resistance to HepG2 and other HCC cell lines, and provided a new significance for improving the effectiveness of DDP in treating HCC.
癌细胞对吉西他滨、奥沙利铂、顺铂、阿霉素和5-氟尿嘧啶等化疗药物产生耐药性是肝癌患者化疗失败的主要原因，尤其是晚期肝癌患者或发生转移的患者。这种新出现的耐药性限制了这些化疗药物的有效性和临床应用。此前的研究报道，耐药肿瘤细胞衍生的外泌体可将其耐药特性转移至某些癌症（包括肺癌和胃癌）中的肿瘤敏感细胞。本研究旨在探究HepG2/DDP细胞衍生的外泌体是否会将顺铂（DDP）耐药性传递给HepG2及其他肝癌敏感细胞，并为临床上使用顺铂进行肝癌护理提供重要指导。
建立HepG2顺铂耐药细胞系（HepG2/DDP），分别从HepG2/DDP和HepG2细胞中分离出外泌体，分别命名为ES-2、ES-1。用DDP或DDP + ES-2处理HepG2或SMMC-7721或Huh7细胞，用ES-1处理HepG2/DDP细胞。然后，通过细胞活力测定和活性氧生成评估HepG2/DDP外泌体转移至HepG2、SMMC-7721和Huh7细胞后耐药性的激活情况。此外，通过蛋白质印迹分析测定P-糖蛋白（P-gp）的相对表达。
成功从顺铂耐药的HepG2细胞中分离出HepG2/DDP细胞衍生的外泌体，并命名为ES-2。与DDP处理组相比，用DDP + ES-2处理的HepG2或SMMC-7721或Huh7细胞的细胞活力增强。此外，与对照组相比，DDP或DDP + ES-2处理的细胞中产生活性氧的浓度显著增加，尽管DDP + ES-2处理组的活性氧浓度明显低于DDP处理组。同时，ES-2表面P-gp的表达上调。
上述结果表明，HepG2/DDP细胞衍生的外泌体赋予了HepG2和其他肝癌细胞系顺铂耐药性，并为提高顺铂治疗肝癌的有效性提供了新的意义。
