茶皂素对肾癌细胞ACHN凋亡与迁移的机制研究  

Mechanism of Tea Saponin on Apoptosis and Migration of Renal Carcinoma Cell Line ACHN

作　　者：田迪 陈志云 吴冲 宋羚 赵英良 桂明英 马啸[1,3] TIAN Di;CHEN Zhi-yun;WU Chong;SONG Ling;ZHAO Ying-liang;GUI Ming-ying;MA Xiao(College of Tea,Yunnan Agricultural University,Kunming 650201,China;Research&Development Center,China Tobacco Yunnan Industrial Co.,Ltd.,Kunming 650231,China;Yunnan Plateau Characteristic Agricultural Industry Research Institute,Kunming 650201,China)

机构地区：[1]云南农业大学茶学院,昆明650201 [2]云南中烟工业有限责任公司技术中心,昆明650231 [3]云南省高原特色农业产业研究院,昆明650201

出　　处：《中国食物与营养》2023年第5期45-50,共6页Food and Nutrition in China

基　　金：云南省重大科技专项-绿色食品国际合作研究中心项目(项目编号:2019ZG009);云南中烟工业有限责任公司技术服务项目(项目编号:2020539200340152);云南省科技项目(项目编号:YCJ 2020-323)。

摘　　要：目的:探究肾癌细胞ACHN经不同浓度茶皂素(TS)组处理后,对细胞凋亡及迁移的影响及其相关机制。方法:以肾癌细胞ACHN为研究对象,在二甲基噻唑蓝比色法(MTT)细胞活性测定后,将细胞培养在含10%胎牛血清的高糖培养基中,分别用0、10、20、30 mg/L浓度的茶皂素处理细胞24 h。对细胞进行苏木精-伊红染色法(HE)、DAPI染色法(DAPI)观察不同茶皂素处理浓度时肾癌细胞的细胞形态及凋亡情况。划痕实验观察不同浓度茶皂素处理情况下细胞的迁移状况,蛋白免疫印迹法对调控细胞凋亡的蛋白,如Bax、Bcl-2、核转录因子-кB(NF-кB P53)、Caspase-9等与对细胞迁移有影响的基质金属蛋白酶-2(MMP-2)蛋白进行印迹分析。结果:在一定范围内,随着茶皂素处理浓度的增加肾癌细胞ACHN凋亡程度逐渐增大,对其迁移能力的抑制逐渐增加,对细胞凋亡有促进作用的Bax、P53、Caspase-9等蛋白的表达与无茶皂素处理组相比有增加趋势,而对细胞凋亡有抑制作用的蛋白Bcl-2表达则呈降低的趋势,促进细胞迁移的MMP-2蛋白表达也受到抑制。结论:茶皂素在肾癌治疗方面具有一定的潜在价值。Objective To investigate the effects of different concentrations of cordycepin on apoptosis and migration of renal cancer ACHN cell lines and the related mechanisms.Method ACHN cells were used as the research object.After MTT cell activity was determined,the cells were cultured in DMEM medium containing 10%fetal bovine serum and treated with 0,10,20,30 mg/L tea saponin for 24 h,respectively.He staining and DAPI staining were performed to observe the cell morphology and apoptosis of renal carcinoma cells treated with different concentrations of tea saponin.The migration status of cells treated with different concentrations of tea saponin was observed by scratch experiment.The proteins that regulate cell apoptosis,such as Bax,Bcl-2,P53,Caspase-9,and MMP-2 proteins of matrix metalloproteinase family,which have influence on cell migration,were analyzed by Western Blot.Result Within a certain range,with the increase of tea saponin treatment concentration,the apoptosis degree of ACHN in renal cancer cells gradually increased,and the inhibition of its migration ability gradually increased.Compared with the group without tea saponin treatment,the expression of Bax,P53,Caspase-9 and other proteins which can promote cell apoptosis showed an increasing trend.The expression of Bcl-2,which can inhibit cell apoptosis,was decreased,and the expression of MMP-2,which can promote cell migration,was also inhibited.Conclusion Tea saponin has potential value in the treatment of renal cancer.

关 键 词：茶皂素 肾癌细胞 细胞活性 凋亡 机制 

分 类 号：R737.11[医药卫生—肿瘤]