二氢杨梅素对人肺腺癌细胞增殖和凋亡的影响及其机制探讨  

Effects of dihydromyricetin on proliferation and apoptosis of human lung adenocarcinoma cells and its mechanism

作　　者：卢平方 梁柱[1] 石伟成[2] 张展飞[1] 蔡小碧[1] 陈春源[1] 林育超[3] 王志刚[1] 

机构地区：[1]广东医学院附属医院心胸外科,广东湛江524001 [2]武警广东省总队医院胸外科,广东广州510507 [3]广东省农垦中心医院胸外科,广东湛江524002

出　　处：《肿瘤》2015年第10期1098-1105,共8页Tumor

摘　　要：目的 :研究二氢杨梅素(dihydromyricetin)对人肺腺癌NCI-H1975细胞增殖和凋亡的影响,并初步探讨其可能的分子作用机制。方法 :采用CCK-8法检测不同浓度(10、25、50、75和100μmol/L)的二氢杨梅素分别作用不同时间(12、24和48 h)对人肺腺癌NCI-H1975细胞增殖的抑制作用。FCM法检测不同浓度(25、50和100μmol/L)的二氢杨梅素作用24 h对肺腺癌NCI-H1975细胞凋亡及细胞周期的影响,同时在倒置相差显微镜下观察NCI-H1975细胞的形态学变化。采用实时荧光定量PCR和蛋白质印迹法分别检测不同浓度(25、50和100μmol/L)的二氢杨梅素作用肺腺癌NCI-H1975细胞24 h后Bcl-2、Bax和Bad等基因的表达水平变化。结果 :二氢杨梅素对人肺腺癌NCI-H1975细胞增殖有明显的抑制作用(P<0.05),且存在时效和量效关系(P值均<0.05)。二氢杨梅素能显著促进肺癌细胞的凋亡(P<0.01),但不诱导细胞周期阻滞(P>0.05)。二氢杨梅素作用后NCI-H1975细胞中Bcl-2 m RNA及其蛋白表达显著下调(P值均<0.05),而Bax m RNA及其蛋白表达显著升高(P值均<0.05);二氢杨梅素作用后Bad m RNA及其蛋白表达无明显变化(P值均>0.05),但Bad蛋白的磷酸化水平显著上调(P<0.05)。结论 :二氢杨梅素可以通过调控线粒体凋亡途径中Bcl-2蛋白家族的表达,介导人肺癌细胞的生长抑制,提示二氢杨梅素可能是一种有效治疗肺癌的潜在药物。Objective To investigate the effects of dihydromyricetin on proliferation and apoptosis of human lung adenocarcinoma cell line NCI-H1975 in vitro, and to explore its molecular mechanism.Methods： After treatment with different concentrations of dihydromyricetin （10, 25, 50, 75 and 100 μmol/L） for different time （12, 24 and 48 h）, the proliferation of human lung adenocarcinoma NCI-H1975 cells was detected by CCK-8 assay. The cell cycle and apoptosis rate of NCI-H1975 cells treated with different concentrations of dihydromyricetin （25, 50 and 100 μmol/L） for 24 h were determined by flow cytometry, and the change of cell morphology was observed by inverted phase contrast microscopy. Real-time fluorescent quantitative-PCR and Western blotting were used to analyze the expression levels of Bcl-2, Bax and Bad genes in NCI-H1975 cells treated with 25, 50 or 100 pmol/L dihydromyricetin for 24 h. Results： Dihydromyricetin significantly inhibited the proliferation of human lung adenocarcinoma NCI-H1975 cells in a dose- and time-dependent manner （P 〈 0.05）. Dihydromyricetin significantly induced apoptosis of NCI-H1975 cells （P 〈 0.01）, but it did not induce cell cycle arrest （P 〉 0.05）. Furthermore, dihydromyricetin significantly decreased the expression levels of Bcl-2 mRNA and protein in NCI-H1975 cells （both P 〈 0.05）, and up-regulated the expression levels of Bax mRNA and protein as well as the expression level of phospho-Bad protein （all P 〈 0.05）, but did not change the expression levels of Bad mRNA and protein （both P 〉 0.05）. Conclusion： Dihydromyricetin can suppress the growth of human lung adenocarcinoma cells through regulating the expression of Bcl-2 protein family in mitochondrial apoptotic pathway. Dihydromyricetin may be a potential drug for the treatment of lung carcinoma.

关 键 词：肺肿瘤 二氢杨梅素 基因 BCL-2 细胞增殖 细胞凋亡 

分 类 号：R284.1[医药卫生—中药学]