摘要
目的:探讨橙皮苷对转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导的人非小细胞肺癌细胞上皮-间质转化的影响。方法:以A549人非小细胞肺癌细胞系作为研究对象。用MTT法,分别检测橙皮苷对正常培养条件或TGF-β1刺激下细胞增殖的影响。应用2种方法评估橙皮苷对TGF-β1刺激条件下A549细胞的上皮间质转化:圆形度值定量评估细胞的形态变化;双抗体夹心酶联免疫吸附法(enzyme linked-immuno-sorbent assay,ELISA)检测上皮细胞钙黏蛋白(E-cadherin,E-cad)、ɑ-平滑肌肌动蛋白(ɑ-smooth muscle actin,ɑ-SMA)、基质金属蛋白酶抑制因子-1(tissue inhibitors of metalloproteinases-1,TIMP-1)和基质金属蛋白酶-9(metallopreoteinases-9,MMP-9)水平。结果:0~40μmol·L-1的橙皮苷对正常培养条件或TGF-β1刺激后A549细胞的增殖没有显著影响。5 ng·mL-1 TGF-β1能够诱导A549细胞的上皮间质转化:由上皮特征的铺路石状的细胞形态转化为梭形形态,圆形度值减少;E-cad分泌量减少和ɑ-SMA合成量增加。另外,MMP-9分泌量增加,MMP-9/TIMP-1比值增加。在细胞出现形态改变后,应用橙皮苷(20μmol·L-1或40μmol·L-1)能够降低ɑ-SMA、MMP-9水平和MMP-9/TIMP-1比值,增加E-cad水平,对细胞的圆形度值没有影响。结论:橙皮苷减轻TGF-β1诱导的非小细胞肺癌细胞的上皮间质转化程度,对非小细胞肺癌的转移和扩散有一定的抑制作用。
OBJECTIVE To study the effects of hesperidin on epithelial mesenchymal transition of human non-small cell cancer cells（A549 cells）induced by TGF-β1.METHODS MTT assay was applied to evaluate cellular proliferation either in normal culture or after stimulation of TGF-β1.Then5 ng·mL-1 TGF-β1 was given to cells and the cellular shape changed from pebble to spindle shape 12 hours after the stimulation.Several doses of hesperidin were given to the cells,respectively.Two methods were applied to evaluate EMT：cellular roundness value for cell morphology,and ELISA for E-cad,ɑ-SMA,MMP-9 and TIMP-1 levels.RESULTS Whether under normal culture conditions or after TGF-β1 stimulation,there was no significant effect on cellular viability in 0～40μmol·L^-1 of hesperidin.EMT of the cells was induced by 5 ng·mL-1 TGF-β1 with cellular morphological changes from cobblestone to spindle shape,cellular roundness value and E-cad level decreased,ɑ-SMA level,MMP-9level and MMP-9/TIMP-1 ratio increased.Hesperidin（20 or 40μmol·L-1）added to the cells after morphological changes resulted in significantly increasedɑ-SMA and MMP-9 levels and MMP-9/TIMP-1 ratio,increased E-cad level,but no significant effect on roundness value.CONCLUSION Hesperidin can alleviate EMT induced by TGF-β1in A549 cells,and inhibit metastasis and diffusion of non-small cell lung cancer cells to certain extent.
作者
任周新
余海滨
李丽
沈俊岭
刘学芳
REN Zhou-xin YU Hai-bin LI Li SHEN Jun-ling LIU Xuefang(Henan University of Traditional Chinese Medicine, Henan Zhengzhou 450046,China Collaborative Innovation Center for Respiratory Disease Diagnosis And Treatment ＆ Chinese Medicine Development of Henan Province, Henan Zhengzhou 450046,China First Affiliated Hospital of Henan University of TCM, Henan Zhengzhou 450000, China Hospital of Henan Academy Institute of Traditional Chinese Medicine, Henan Zhengzhou 450000, China)
出处
《中国医院药学杂志》
CAS
北大核心
2017年第8期690-693,共4页
Chinese Journal of Hospital Pharmacy
基金
国家自然科学基金项目(编号:81473650)
河南省高等学校重点科研项目计划(编号:16A360002)
关键词
橙皮苷
非小细胞肺癌细胞
增殖
上皮间质转化
hesperidin
non-small cell lung cancer（NSCLC）
proliferation
epithelial mesenchymal transition（EMT）
