扶正抗癌方对吉非替尼在肺癌H1650、A549细胞上的增效作用  

The gefitinib-sensitivity enhancing effects of FZKA decoction on H1650 and A549 cells via MET/PI3K

作　　者：杨小兵[1] 庄媛媛 龙顺钦[1] 周宇姝[1] 黎金华 吴万垠[1] YANG Xiao-bing;ZHUANG Yuan-yuan;LONG Shun-qin;ZHOU Yu-shu;LI Jin-hua;WU Wan-yin(Department of Oncology,Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510370,Guangdong,China)

机构地区：[1]广东省中医院肿瘤科,广东广州510370 [2]广东省中医院口腔科,广东广州510370 [3]广州中医药大学第二临床医学院,广东广州510006

出　　处：《广东医学》2018年第20期3002-3009,共8页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81503507;81273965);广东省自然科学基金资助项目(编号:2015A030310245);广东省科学技术厅-广东省中医药科学院联合科研项目(编号:2016A020226035)

摘　　要：目的观察扶正抗癌(FZKA)方联合吉非替尼(Gefitinib)对非小细胞肺癌H1650、A549细胞增殖的影响,并探讨其作用的分子机制。方法用空白对照组、FZKA方组(0. 4、0. 8、1. 2、1. 6、2. 0、2. 4、2. 8、3. 2mg/m L)、Gefitinib组(2、4、8、10、15、20、25μmol/L)、FZKA方(1. 6 mg/m L)联合Gefitinib组(2、4、8、10、15、20、25μmol/L,联合用药组)分别干预细胞后,采用MTT法分别检测其对H1650、A549细胞活力的影响,分别计算出半数致死率(IC50); q PCR法检测空白对照组、FZKA方组、Gefitinib组及联合用药组原癌基因c-MET转录水平(mRNA)的变化; Western blot法分析MET、p-Met(Tyr1234/1235)、PI3K蛋白的表达。结果与空白对照组相比,FZKA方、Gefitinib能显著抑制H1650、A549细胞增殖并呈浓度依赖性。与Gefitinib单药相比,FZKA方联合Gefitinib的抑制效果优于Gefitinib单药,FZKA方对Gefitinib具有增敏作用。与空白对照组相比,FZKA方以浓度依赖性下调MET蛋白的表达(P <0. 05,P <0. 01),以时间依赖性下调p-MET (Tyr1234/1235)蛋白的表达(P <0. 05,P <0. 01)。FZKA方联合Gefitinib下调MET mRNA和MET、PI3K蛋白表达效果优于FZKA方、Gefitinib单独用药(P <0. 05)。结论 FZKA方对Gefitinib起到增敏作用,这个作用可能是通过调控MET/PI3K通路来实现的。Objective To observe the Gefitinib-sensitivity enhancing effects and its molecular mechanism of FZKA decoction on H1650 and A549 cells. Methods The methyl-thiazolyl-tetrazolium（ MTT） assay was used to detect the effects of FZKA decoction on cell proliferation and apoptosis. H1650 and A549 cells were assigned into blank group,FZKA decoction groups（ 0. 4,0. 8,1. 2,1. 6,2. 0,2. 4,2. 8 and 3. 2 mg/m L）,Gefitinib groups（ 2,4,8,10,15,20 and 25 μmol/L） and FZKA decoction（ 1. 6 mg/m L） combined with gefitinib groups（ 2,4,8,10,15,20 and 25μmol/L）. The quantitative real-time PCR（ q PCR） was used to detect the MET mRNA. Western blot assay was used to detect the expression levels of phosphatase and tensin homolog deleted on chromosome ten（ MET）,Phospho-MET（ p-MET） and phosphatidylinositol-3-kinase（ PI3 K）. The correlation analysis was performed. Results As compared with the blank group,the proliferation of H1650 and A549 cells were significantly inhibited by FZKA decoction in a concentration-dependent manner（ P〈0. 01）,and the inhibiting effect of combination of FZKA decoction and gefitinib was significantly better than FZKA decoction alone（ P〈0. 01）. As compared with the blank group,the mRNA and protein expression of MET were upregulated in a concentration-dependent manner（ P〈0. 05）. The expression of MET and PI3 K was significantly lower in FZKA decoction and gefitinib combination than gefitinib alone（ P〈0. 05）. Conclusion FZKA decoction enhances the gefitinib-sensitivity of H1650 and A549 cells via MET down-regulation.

关 键 词：扶正抗癌方 吉非替尼 增敏 MET PI3K 

分 类 号：R734.2[医药卫生—肿瘤]