小豆蔻素对黑色素瘤细胞的作用及机制  

Effects and mechanism of cardamom on melanoma cells

作　　者：张芬 戴耕武[1] 罗东升[1] 赖小梅[2] 杨镓宁[1] ZHANG Fen;DAI Geng-wu;LUO Dong-sheng;LAI Xiao-mei;YANG Jia-ning(Institute of Dermatology,Sichuan Academy of Medical Sciences.Sichuan Provincial People's Hospital,Chengdu 610072,Sichuan Province,China;Department of Dermatology,Sichuan Academy of Medical Sciences.Sichuan Provincial People's Hospital,Chengdu 610072,Sichuan Province,China)

机构地区：[1]四川省医学科学院·四川省人民医院皮研所,四川成都610072 [2]四川省医学科学院·四川省人民医院皮肤外科,四川成都610072

出　　处：《中国临床药理学杂志》2023年第22期3286-3290,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探究小豆蔻素对黑色素瘤B16-F10细胞增殖、凋亡、迁移和侵袭及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素(mTOR)信号通路的影响。方法B16-F10细胞分为对照组(B16-F10细胞未做任何处理),低、中、高剂量实验组(10、20和40μmol·L^(-1)小豆蔻素),LY294002组(20μmol·L^(-1)LY294002),高剂量+740Y-P组(40μmol·L^(-1)小豆蔻素+20μmol·L^(-1)740Y-P)。噻唑蓝(MTT)法检验B16-F10细胞的增殖能力;Annexin-V-FITC/PI双染法检测B16-F10细胞的凋亡率;划痕愈合实验检测B16-F10细胞的迁移能力;Transwell实验检测B16-F10细胞的侵袭能力;蛋白印迹法检测蛋白表达。结果对照组、高剂量实验组、LY294002组和高剂量+740Y-P组的细胞增殖率分别为(100.00±1.95)%、(39.59±3.75)%、(42.34±4.02)%和(91.32±11.34)%,细胞凋亡率分别为(6.97±0.59)%、(19.21±1.95)%、(19.02±2.02)%和(7.68±0.74)%,划痕愈合率分别为(67.85±5.29)%、(21.35±3.48)%、(24.02±3.70)%和(63.56±5.12)%,侵袭细胞数分别为(150.48±14.32)、(53.12±5.01)、(52.34±5.14)和(128.32±11.89)个,p-PI3K/PI3K分别为0.80±0.08、0.49±0.06、0.50±0.04和0.73±0.09,p-Akt/Akt分别为0.81±0.08、0.46±0.04、0.45±0.05和0.71±0.08,p-mTOR/mTOR分别为0.77±0.05、0.35±0.03、0.36±0.04和0.69±0.07。上述指标,高剂量实验组、LY294002组与对照组比较,高剂量+740Y-P组与高剂量实验组比较,差异均有统计学意义(均P<0.05)。结论豆蔻素可能通过抑制PI3K/Akt/mTOR信号通路抑制黑色素瘤B16-F10细胞增殖、迁移和侵袭,促进其凋亡。Objective To investigate the effects of cardamonin on the proliferation,apoptosis,migration,and invasion of melanoma B16-F10 cells and the phosphatidylinositol 3 kinase(PI3K)/serine/threonine kinase(Akt)/mammalian target of rapamycin(mTOR)signaling pathway.Methods B16-F10 cells cultured in vitro and divided into:Control group(B16-F10 cells without any treatment),experimental-L,-M,-H groups(10,20,40μmol·L^(-1)cardamom),LY294002 group(20μmol·L^(-1)LY294002),experimental-H+740Y-P group(40μmol·L^(-1)cardamonin+20μmol·L^(-1)740Y-P).Methyl thiazolyl tetrazolium(MTT)assay were applied to examine the proliferation ability of B16-F10 cells;Annexin-V-FITC/PI double staining was applied to detect the apoptosis rate of B16-F10 cells;the migration ability of B16-F10 cells was detected by scratch healing assay;Transwell assay was applied to detect the invasion abilities of B16-F10 cells;Western blot was applied to detect expression of proteins.Results The cell proliferation rates of control group,experimental-H group,LY294002 group and experimental-H+740Y-P group were(100.00±1.95)%,(39.59±3.75)%,(42.34±4.02)%and(91.32±11.34)%,respectively;the apoptosis rates were(6.97±0.59)%,(19.21±1.95)%,(19.02±2.02)%and(7.68±0.74)%,respectively;scratch healing rates were(67.85±5.29)%,(21.35±3.48)%,(24.02±3.70)%and(63.56±5.12)%,respectively;the number of invasive cells were(150.48±14.32),(53.12±5.01),(52.34±5.14)and(128.32±11.89),respectively;p-PI3K/PI3K ratios were 0.80±0.08,0.49±0.06,0.50±0.04 and0.73±0.09,respectively;p-Akt/Akt ratios were 0.81±0.08,0.46±0.04,0.45±0.05 and 0.71±0.08,respectively;p-m TOR/m TOR ratios were 0.77±0.05,0.35±0.03,0.36±0.04 and 0.69±0.07,respectively.The experimental-H groups,the LY294002 group were compared with the control group,the Experimental-H+740Y-P group and the experimental-H group were compared,and the differences of the obove indicators were statistically significant(P<0.05).Conclusion Cardamonin may inhibit the proliferation,migration and invasion of melanoma B16-F1

关 键 词：小豆蔻素 黑色素瘤 磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路 增殖 迁移 侵袭 

分 类 号：R28[医药卫生—中药学]