穗花杉双黄酮对鼻咽癌细胞恶性生物学行为的影响  

Impacts of amentoflavone on the malignant biological behavior of nasopharyngeal carcinoma cells

作　　者：杨西国 杨西霞 朱婷 李莉 YANG Xi-guo;YANG Xi-xia;ZHU Ting;LI Li(Department of Otolaryngology,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan 271100,Shandong Province,China;Department of Pharmacy,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan 271100,Shandong Province,China;Department of Internal Medicine,the Eighth People’s Hospital of Jinan,Jinan 271100,Shandong Province,China)

机构地区：[1]济南市中西医结合医院耳鼻喉科,山东济南271100 [2]济南市中西医结合医院药剂科,山东济南271100 [3]济南市第八人民医院内科,山东济南271100

出　　处：《中国临床药理学杂志》2023年第21期3111-3115,共5页The Chinese Journal of Clinical Pharmacology

基　　金：山东省中医药科技基金资助项目(Z-2022003)。

摘　　要：目的研究穗花杉双黄酮(AF)调节Ras/Raf/促分裂原活化蛋白激酶激酶(MEK)/细胞外信号调节激酶(ERK)信号通路对鼻咽癌(NPC)细胞恶性生物学行为的影响。方法将NPC细胞株CNE-1随机分为对照组、低浓度AF组(AF-L组,50μmol·L^(-1)AF)、中浓度AF组(AF-M组,75μmol·L^(-1)AF)、高浓度AF组(AF-H组,125μmol·L^(-1)AF)和高浓度AF+Ras激活剂RASAL1组(AF-H+RASAL1组,125μmol·L^(-1)AF+2 ng·mL^(-1)RASAL1)。以噻唑蓝(MTT)实验检测细胞增殖能力;以划痕愈合实验检测细胞迁移能力;以Transwell实验检测细胞侵袭能力;以流式细胞术检测细胞凋亡率;以蛋白质印迹法检测细胞中Ras/Raf/MEK/ERK信号通路相关蛋白表达。结果对照组、AF-M组、AF-H组CNE-1细胞OD490值(48 h)为0.62±0.06、0.45±0.05、0.29±0.03;细胞划痕愈合率为(38.66±4.07)%、(23.14±2.20)%、(12.47±1.15)%;细胞侵袭数目为(137.59±10.25)、(103.42±8.49)、(76.38±7.96)个;Ras蛋白表达水平分别为0.84±0.08、0.59±0.06、0.34±0.03;Raf蛋白表达水平分别为0.79±0.08、0.53±0.05、0.24±0.02;MEK蛋白表达水平分别为0.87±0.09、0.64±0.06、0.28±0.03;ERK1/2磷酸化水平分别为0.75±0.08、0.50±0.05、0.21±0.02;细胞凋亡率分别为(4.25±0.63)%、(18.42±2.10)%、(35.28±2.26)%,以上指标,AF-M组、AF-H组与对照组比较,差异均有统计学意义(均P<0.05)。RASAL1减弱了AF对NPC细胞增殖、迁移和侵袭的抑制作用,抑制了其凋亡。结论AF可能通过下调Ras/Raf/MEK/ERK信号通路抑制NPC细胞增殖、迁移和侵袭,促进其凋亡。Objective To explore the offect of amentoflavone(AF)on the malignant biological behavior of nasopharyngeal carcinoma(NPC)cells by regulating the Ras/Raf/mitogen activated protein kinase(MEK)/extracellular signal regulated kinase(ERK)signaling pathway.Methods NPC cell line CNE-1 was randomly grouped into control group,low concentration AF group(AF-L group,50μmol·L^(-1) AF),medium concentration AF group(AF-M group,75μmol·L^(-1) AF),high concentration AF group(AF-H group,125μmol·L^(-1) AF),high concentration AF+Ras activator RASAL1 group(AF-H+RASAL1 group,125μmol·L^(-1) AF+2 ng·mL^(-1) RASAL1).MTT cytotoxicity experiment was applied to detect the proliferation ability of cells in each gro up;wound healing test was applied to detect cell migration ability;Transwell experiment was applied to detect cell invasion ability;flow cytometry was applied to detect cell apoptosis rate;Western Blot was applied to detect the expression of Ras/Raf/MEK/ERK signaling pathway related proteins in cells.Results The OD490 values(48 h)of CNE-1 cells in control group,AF-M group and AF-H group were 0.62±0.06,0.45±0.05 and 0.29±0.03;cell scratch healing rates were(38.66±4.07)%,(23.14±2.20)%,(12.47±1.15)%;the number of cells invaded were 137.59±10.25,103.42±8.49,76.38±7.96;Ras protein expression levels were 0.84±0.08,0.59±0.06,0.34±0.03;Raf protein expression levels were 0.79±0.08,0.53±0.05,0.24±0.02;the expression levels of MEK protein were 0.87±0.09,0.64±0.06,0.28±0.03,respectively;the phosphorylation levels of ERK1/2 were 0.75±0.08,0.50±0.05 and 0.21±0.02;the apoptosis rates were(4.25±0.63)%,(18.42±2.10)%and(35.28±2.26)%,respectively.There were statistically significant differences between AF-M group,AF-H group and control group(all P<0.05).RASAL1 weakened the inhibitory effects of AF on the proliferation,migration,and invasion of NPC cells,and inhibited their apoptosis.Conclusion AF may inhibit the proliferation,migration,and invasion of NPC cells and promote their apoptosis by down-regulating the Ras/Ra

关 键 词：穗花杉双黄酮 Ras/Raf/促分裂原活化蛋白激酶激酶/细胞外信号调节激酶信号通路 鼻咽癌 增殖 迁移 侵袭 凋亡 

分 类 号：R28[医药卫生—中药学]