目的:电化学疗法,临床建立用于治疗(皮下)皮肤肿瘤,已在欧洲标准电化学疗法操作程序(ESOPE)的框架内标准化。由于化疗药物的常见副作用,最近的进展集中在非细胞毒性药物上,像钙,诱导细胞死亡(钙电穿孔)。因此,这项研究旨在确定博来霉素或顺铂的电化学疗法的疗效,或使用ESOPE方案在体外对人肝细胞癌细胞(HepG2)进行钙电穿孔。
方法:在化疗药物博来霉素或顺铂(0-20µM)进行电化学化疗后,用MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物)测定HepG2细胞活力,或钙电穿孔后(0-20mM),使用ESOPE方案(8个矩形脉冲,1000V/cm,100µs)与非电穿孔药物治疗相比。
结果:电穿孔样品中细胞活力显著降低,与未电穿孔的对照相比(差异27-75%)。博来霉素和钙电穿孔的电化学疗法,达到(几乎)完全细胞死亡(-1±3%和2.5±2%),在2.5µM和2.5mM的最低浓度下,分别。用2.5µM顺铂进行电化学治疗,显着降低细胞活力仅68%(±7%)。
结论:博来霉素或顺铂的电化学疗法,与单独的非电穿孔药物处理相比,使用ESOPE方案,或钙电穿孔在体外降低HepG2细胞活力方面更有效。当比较电化学时,HepG2细胞对博莱霉素比顺铂更敏感,浓度相似。钙电穿孔与博来霉素的电化学疗法具有相同的效果,但钙可能具有更好的安全性和一些治疗优势。
OBJECTIVE: Electrochemotherapy, clinically established for treating (sub)cutaneous tumors, has been standardized in the framework of the European Standard Operating Procedure on
Electrochemotherapy (ESOPE). Due to common side effects of chemotherapeutic drugs, recent advances focus on non-cytotoxic agents, like calcium, to induce cell death (calcium electroporation). Therefore, this study aims to determine the efficacy of
electrochemotherapy with bleomycin or cisplatin, or calcium electroporation on human hepatocellular carcinoma cells (HepG2) in vitro using the ESOPE protocol.
METHODS: HepG2 cell viability was measured with a MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay after electrochemotherapy with the chemotherapeutic drugs bleomycin or cisplatin (0-20 µM), or after calcium electroporation (0-20 mM), to determine its efficacy on HepG2 cells in vitro using the ESOPE protocol (8 rectangular pulses, 1000 V/cm, 100 µs) compared to non-electroporated drug treatment.
RESULTS: Cell viability was significantly lower in electroporated samples, compared to their non-electroporated controls (27-75% difference).
Electrochemotherapy with bleomycin and calcium electroporation, reached (almost) complete cell death (- 1 ± 3% and 2.5 ± 2%), in the lowest concentration of 2.5 µM and 2.5 mM, respectively.
Electrochemotherapy with 2.5 µM cisplatin, significantly decreased cell viability to only 68% (± 7%).
CONCLUSIONS: Electrochemotherapy with bleomycin or cisplatin, or calcium electroporation were more effective in reducing the HepG2 cell viability in vitro using the ESOPE protocol compared to the non-electroporated drug treatments alone. When comparing electrochemotherapy, HepG2 cells are more sensitive to bleomycin than cisplatin, in similar concentrations. Calcium electroporation has the same effectiveness as electrochemotherapy with bleomycin, but calcium potentially has a better safety profile and several treatment advantages.