Department of Gynecology, YanTaiShan Hospital, YanTai City, China,
Chemotherapy. 2019;64(3):119-128. doi: 10.1159/000501633. Epub 2019 Oct 29.
To investigate whether TRIAP1inhibition affects the ovarian cancer cell resistance to cisplatin (DDP) via the Cyt c/Apaf-1/caspase-9 pathway by in vitro and in vivo experiments.
CCK8 assay was performed to find out how treatment with both TRIAP1 siRNA and DDP affects the cell viability of SKOV3 cells and DDP-resistant human ovarian carcinoma cell line SKOV3/DDP. SKOV3/DDP cells were transfected with control siRNA or TRIAP1 siRNA before 24 h of treatment with DDP (5 μg/mL). Flow cytometry was employed to detect cell apoptosis and Western blot to examine the expressions of Cyt c/Apaf-1/caspase-9 pathway-related proteins. SKOV3/DDP cells transfected with control siRNA or TRIAP1 siRNA were subcutaneously injected into BALB/c-nu/nu nude mice followed by the intraperitoneal injection of DDP (4 mg/kg). Cyt c/Apaf-1/caspase-9 pathway in transplanted tumors was detected by immunohistochemistry.
TRIAP1 expression declined in SKOV3 cells when compared with SKOV3/DDP cells. The proliferation rate was lower in SKOV3/DDP cells transfected with TRIAP1 siRNA combined with treatment of DDP (1, 2, 4, 6, 8, 16, 32 μg/mL) than in those transfected with control siRNA. Moreover, the TRIAP1 siRNA group had an increased SKOV3/DDP cell apoptosis rate with the activation of the Cyt c/Apaf-1/caspase-9 pathway. During DDP treatment, nude mice in TRIAP1 siRNA group had slower growth and smaller size of transplanted tumor than those in control siRNA group, with increased expression of Cyt c, Apaf-1, and caspase-9.
TRIAP1 inhibition may enhance the sensitivity of SKOV3/DDP cells to cisplatin via activation of the Cyt c/Apaf-1/caspase-9 pathway.
通过体外和体内实验研究 TRIAP1 抑制是否通过 Cyt c/Apaf-1/caspase-9 通路影响卵巢癌细胞对顺铂（DDP）的耐药性。
采用 CCK8 法检测 TRIAP1 siRNA 联合 DDP 处理对 SKOV3 细胞和耐 DDP 的人卵巢癌细胞系 SKOV3/DDP 细胞活力的影响。用 DDP（5 μg/ml）预处理 SKOV3/DDP 细胞 24 h 前，用对照 siRNA 或 TRIAP1 siRNA 转染 SKOV3/DDP 细胞。流式细胞术检测细胞凋亡，Western blot 检测 Cyt c/Apaf-1/caspase-9 通路相关蛋白表达。将对照 siRNA 或 TRIAP1 siRNA 转染的 SKOV3/DDP 细胞皮下注射到 BALB/c-nu/nu 裸鼠体内，然后腹腔注射 DDP（4 mg/kg）。免疫组化法检测移植瘤中 Cyt c/Apaf-1/caspase-9 通路。
与 SKOV3/DDP 细胞相比，SKOV3 细胞中 TRIAP1 表达下降。与转染对照 siRNA 的 SKOV3/DDP 细胞相比，转染 TRIAP1 siRNA 并联合 DDP（1、2、4、6、8、16、32 μg/ml）处理的 SKOV3/DDP 细胞增殖率较低。此外，TRIAP1 siRNA 组 SKOV3/DDP 细胞凋亡率升高，Cyt c/Apaf-1/caspase-9 通路激活。在 DDP 处理过程中，TRIAP1 siRNA 组裸鼠移植瘤生长较慢，体积较小，Cyt c、Apaf-1 和 caspase-9 表达增加。
TRIAP1 抑制可能通过激活 Cyt c/Apaf-1/caspase-9 通路增强 SKOV3/DDP 细胞对顺铂的敏感性。
