Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Kasr El-Aini St., Cairo Postal Code: 11562, Egypt.
Department of Medical Biochemistry, National Cancer Institute, Cairo University, Kasr El-Aini st., Fom El-Khalig, Cairo Postal Code: 11796, Egypt.
Life Sci. 2022 Jul 15;301:120594. doi: 10.1016/j.lfs.2022.120594. Epub 2022 Apr 29.
Aim Hepatocellular carcinoma (HCC) is one of the leading causes of cancer death worldwide. Systemic chemotherapy such as cisplatin and multi-targeted receptor tyrosine kinase inhibitors, including sunitinib, has marginal activity and frequent toxicity. Recently, necroptosis has been investigated as a potential target in treating cancer. Our aim is to evaluate the influence of cisplatin-sunitinib combination on HepG2 cells regarding their cytotoxicity and implicated intracellular pathways.
The half-maximal inhibitory concentration (IC) values of cisplatin, sunitinib, and their combination were determined by Sulforhodamine-B assay. Bcl-2 and Bax protein levels were assayed using western blot. ELISA technique was used to measure pRIPK3/RIPK3, pERK/ERK, caspase-9, caspase-8, malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GPx).
Cisplatin-sunitinib combination exhibited a superior cytotoxic effect on HepG2 cells. Low concentrations of 4 μg/ml cisplatin and 2.8 μg/ml sunitinib showed significant Bcl-2 down-regulation and Bax up-regulation. The combined treatment also lowered pRIPK3/RIPK3 by 74% (p < 0.05) compared to the control. Significant increase in pERK/ERK by 3.9 folds over the normal control was also demonstrated. Moreover, combined treatment produced a significant 4 and 4.6 folds increase in caspase-9 and -8 levels. An increase in MDA level by 1.3 folds, a decrease in the intracellular GSH level by 63%, and an increase in GPx level by 1.17 folds were demonstrated.
Sunitinib modulated cisplatin effect on cytotoxicity, oxidative stress, apoptosis, necroptosis and MAPK pathways. Sunitinib enhanced cisplatin-induced apoptosis and increased oxidative stress, but decreased necroptosis. Combined cisplatin and sunitinib might be promising for treating advanced HCC.
目的 肝细胞癌（HCC）是全球癌症死亡的主要原因之一。顺铂等系统化疗和多靶点受体酪氨酸激酶抑制剂，包括舒尼替尼，活性有限，毒性频繁。最近，细胞坏死已被研究作为治疗癌症的潜在靶点。我们的目的是评估顺铂-舒尼替尼联合应用对 HepG2 细胞的细胞毒性及其涉及的细胞内途径的影响。
材料和方法 通过磺酰罗丹明 B 测定法确定顺铂、舒尼替尼及其组合的半最大抑制浓度（IC）值。使用 Western blot 测定 Bcl-2 和 Bax 蛋白水平。使用 ELISA 技术测量 pRIPK3/RIPK3、pERK/ERK、caspase-9、caspase-8、丙二醛（MDA）、谷胱甘肽（GSH）和谷胱甘肽过氧化物酶（GPx）。
主要发现 顺铂-舒尼替尼联合应用对 HepG2 细胞表现出优越的细胞毒性作用。低浓度的 4μg/ml 顺铂和 2.8μg/ml 舒尼替尼显示出明显的 Bcl-2 下调和 Bax 上调。与对照组相比，联合治疗还使 pRIPK3/RIPK3 降低了 74%（p<0.05）。还证明了 pERK/ERK 比正常对照组增加了 3.9 倍。此外，联合治疗还使 caspase-9 和 -8 水平分别增加了 4 倍和 4.6 倍。MDA 水平增加了 1.3 倍，细胞内 GSH 水平降低了 63%，GPx 水平增加了 1.17 倍。
意义 舒尼替尼调节了顺铂对细胞毒性、氧化应激、细胞凋亡、细胞坏死和 MAPK 途径的作用。舒尼替尼增强了顺铂诱导的细胞凋亡，并增加了氧化应激，但减少了细胞坏死。联合顺铂和舒尼替尼可能对治疗晚期 HCC 有希望。
