Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
College of Chinese Medicine, School of Post-Baccalaureate Chinese Medicine, China Medical University, Taichung, Taiwan, R.O.C.
In Vivo. 2022 Mar-Apr;36(2):582-595. doi: 10.21873/invivo.12741.
BACKGROUND/AIM: Lung cancer notably contributes to tumor-associated mortality worldwide, and standard chemotherapy is used for lung cancer patients. However, its therapeutic efficacy remains unsatisfactory. This study aimed to evaluate the effects and molecular mechanisms of sorafenib and bufalin combination therapy on lung cancer cells in vitro.
NCI-H292 cells were treated with sorafenib, bufalin, and sorafenib in combination with bufalin. Cell viability, ROS production, Ca release, and mitochondrial membrane potential were examined by flow cytometric assay. Annexin V/PI staining and chromatin condensation were examined by the apoptosis assays. Finally the molecular mechanism of apoptosis-associated protein expression was investigated by western blotting.
NCI-H292 cells treated with sorafenib in combination with bufalin showed significantly decreased viability, enhanced cellular apoptosis, and DNA condensation when compared to that with sorafenib or bufalin alone. Moreover, the combination treatment exhibited higher reactive oxygen species (ROS) production and lower mitochondrial membrane potential (ΔΨm). The combined treatment resulted in higher expression of SOD but lower catalase compared to sorafenib treatment alone. Compared to sorafenib or bufalin treatment alone, the combination treatment resulted in lower Bcl-2 expression but higher Bax, Bad, APAF-1, caspase-3, and caspase-9.
Sorafenib in combination with bufalin shows more potent cytotoxic effects and cell apoptosis than sorafenib or bufalin treatment alone in NCI-H292 cells. The combined treatment significantly enhanced apoptotic cell death in NCI-H292 lung cancer cells by activating ROS-, mitochondria-, and caspase-signaling pathways in vitro.
背景/目的：肺癌是全球肿瘤相关死亡的主要原因，标准的化疗用于肺癌患者。然而，其治疗效果仍不尽人意。本研究旨在评估索拉非尼和蟾毒灵联合治疗对体外肺癌细胞的作用及其分子机制。
用索拉非尼、蟾毒灵和索拉非尼联合蟾毒灵处理 NCI-H292 细胞。通过流式细胞术检测细胞活力、ROS 产生、Ca 释放和线粒体膜电位。通过凋亡检测检测 Annexin V/PI 染色和染色质浓缩。最后通过 Western blot 研究凋亡相关蛋白表达的分子机制。
与单独使用索拉非尼或蟾毒灵相比，用索拉非尼联合蟾毒灵处理的 NCI-H292 细胞显示出明显降低的活力、增强的细胞凋亡和 DNA 浓缩。此外，联合治疗表现出更高的活性氧（ROS）产生和更低的线粒体膜电位（ΔΨm）。与单独使用索拉非尼治疗相比，联合治疗导致 SOD 表达增加而过氧化氢酶表达降低。与单独使用索拉非尼或蟾毒灵治疗相比，联合治疗导致 Bcl-2 表达降低而 Bax、Bad、APAF-1、caspase-3 和 caspase-9 表达升高。
与单独使用索拉非尼或蟾毒灵相比，索拉非尼联合蟾毒灵在 NCI-H292 细胞中表现出更强的细胞毒性作用和细胞凋亡作用。联合治疗通过激活 ROS-、线粒体-和 caspase 信号通路，显著增强了 NCI-H292 肺癌细胞的凋亡性细胞死亡。
