摘要
目的观察体外联用青蒿琥酯对顺铂抗前列腺癌活性的影响并研究其机制。方法人前列腺癌细胞系LNCaP分为对照组、青蒿琥酯组、顺铂组、顺铂+青蒿琥酯组、顺铂+青蒿琥酯+MET质粒组。采用噻唑蓝(MTT)法检测前列腺癌细胞系LNCaP的细胞活力抑制率;流式细胞术检测LNCaP细胞的凋亡;Western blot实验检测LNCaP细胞Met表达水平,AKT和Bad磷酸化水平,caspase-9和caspase-3活化水平;免疫共沉淀法检测Bad与Bcl-xl及Bcl-2相互作用。结果顺铂+青蒿琥酯组LNCaP细胞活力抑制率(61.4±4.9)%和凋亡率(34.8±2.9)%均高于顺铂单处理组的细胞活力抑制率(14.7±1.1)%和凋亡率(12.5±1.0)%(P均<0.05)。顺铂+青蒿琥酯组LNCaP细胞Met表达水平,AKT和Bad磷酸化水平均低于顺铂组,而顺铂+青蒿琥酯组LNCaP细胞Bad与Bcl-xl及Bcl-2结合水平,caspase-9和caspase-3活化水平均高于顺铂组(P均<0.05)。青蒿琥酯发挥对顺铂的辅助治疗作用依赖于Met的抑制,顺铂+青蒿琥酯+Met质粒组LNCaP的细胞活力抑制率(20.3±1.4)%和凋亡率(15.5±1.2)%均低于顺铂+青蒿琥酯组的细胞活力抑制率(61.4±4.9)%和凋亡率(34.8±2.9)%(P均<0.05)。结论青蒿琥酯可能通过Met/AKT/Bad途径增强顺铂对前列腺癌细胞的杀伤活性。
Objective To observe the effect of artesunate on the anti-prostate cancer activity of cisplatin in vitro and to study its mechanism. Methods Human prostate cancer cell line LNCaP was divided into control group, artesunate group, cisplatin group, cisplatin+artesunate group, cisplatin + artesunate + MET plasmid group. MTT assay was used to detect the cell viability inhibition rate of LNCaP cells;flow cytometry was performed to detect the apoptosis of cells;Western blot was conducted to detect the expression of Met, AKT and Bad phosphorylation levels, caspase-9 and caspase-3 activation levels;co-immunoprecipitation was used to test the interaction between Bad and Bcl-xl and Bcl-2. Results The inhibition rate of LNCaP cell viability[(61.4±4.9)%] and apoptotic rate [(34.8±2.9)% in the cisplatin + artesunate group were higher than those in the cisplatin group[(14.7±1.1)% and (12.5±1.0)%, respectively], all P<0.05. The levels of Met expression, AKT and Bad phosphorylation in the cisplatin + artesunate group were lower than those in the cisplatin group, while the interaction of Bad with Bcl-xl/Bcl-2, and activation of caspase-9 and caspase-3 in the cisplatin + artesunate group were higher than those in the cisplatin group, all P<0.05. The adjuvant effect of artesunate on cisplatin was dependent on the inhibition of Met. The inhibition rate of cell viability[(20.3±1.4)%] and apoptosis rate[(15.5±1.2)%] of cells in the group of cisplatin + artesunate + Met plasmid were lower than those in the cisplatin + artesunate group[(61.4±4.9)% and (34.8±2.9)%, respectively], all P<0.05. Conclusion Artesunate could enhance the cisplatin-induced cytotoxicity against prostate cancer cells via the Met/AKT/Bad pathway.
作者
周丽娜
ZHOU Lina(Department of Medical Oncology, Tongde Hospital of Zhejiang province, Hangzhou, Zhejiang province, 310000, China)
出处
《浙江中西医结合杂志》
2019年第6期446-450,共5页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
