细胞间充质-上皮转化因子在口腔舌鳞癌吉非替尼获得性耐药中的作用研究

Role of cellular-mesenchymal to epithelial transition factor on gefitinib acquired drug resistance in tongue squamous cell carcinoma

目的探讨细胞间充质-上皮转化因子(c-MET)在人舌鳞癌细胞吉非替尼获得性耐药中的作用及机制。方法体外培养CAL27R细胞并分为空白对照组、对照组、实验组和联合组。空白对照组给予二甲基亚砜;对照组给予5μmol·L^(-1)吉非替尼;实验组给予5μmol·L^(-1)SU11274;联合组给予5μmol·L^(-1)吉非替尼+5μmol·L^(-1)SU11274。4组均干预48 h。用噻唑蓝法检测细胞的增殖能力,用流式细胞术检测细胞的凋亡情况,用Western blot检测蛋白的表达情况。结果空白对照组、对照组、实验组和联合组的细胞存活率分别为(100.00±0)%,(96.53±0.35)%,(73.62±2.52)%和(43.13±8.51)%,细胞凋亡率分别为(7.12±1.30)%,(10.49±2.38)%,(12.32±1.75)%和(49.67±4.76)%,磷酸化蛋白激酶B蛋白相对表达量分别为1.13±0.01,1.02±0.04,0.94±0.01和0.74±0.01,蛋白激酶B蛋白相对表达量分别为2.45±0.70,2.20±0.05,1.74±0.02和0.96±0.02,磷酸化细胞外调节蛋白激酶蛋白相对表达量分别为1.42±0.06,1.40±0.02,0.99±0和0.91±0.02,细胞外调节蛋白激酶蛋白相对表达量分别为1.53±0.17,1.37±0.10,1.26±0.10和0.90±0.01,实验组和联合组的上述指标与空白对照组比较,差异均有统计学意义(均P <0.05)。结论 c-MET及其下游信号通路的激活是舌鳞癌细胞对吉非替尼获得性耐药形成的重要机制,可以将c-MET作为克服获得性耐药的潜在治疗靶点。

Objective To investigate the role and mechanism of cellular-mesenchymal to epithelial transition factor(c-MET) in gefitinib acquired drug resistance of human tongue squamous cell carcinoma.Methods CAL27 R cells was cultured in vitro and divided into blank control,control,experimental and combination groups.Blank control group was treated with dimethyl sulfoxide.Control group was treated with5 μmol·L^(-1) gefitinib.Experimental group was treated with 5 μmol·L^(-1)SU11274.Combination group was treated with 5 μmol·L^(-1) gefitinib and5 μmol·L^(-1) SU11274 for 48 h.Methyl thiazolyl tetrazolium assay was used to detect the proliferation of cells.Flow cytometry was used to detect apoptosis.Western Blot assay was used to detect the expressions level of protein.Results The cell survival rates of blank control,control,experimental and combination groups were(100.00±0) %,(96.53±0.35) %,(73.62±2.52) % and(43.13±8.51) %,the apoptosis rates were(7.12±1.30) %,(10.49±2.38) %,(12.32±1.75) % and(49.67±4.76) %,the relative expression levels of phosphorylated protein kinase B were 1.13±0.01,1.02±0.04,0.94±0.01 and 0.74±0.01,the relative expression levels of protein kinase B were 2.45±0.70,2.20±0.05,1.74±0.02 and 0.96±0.02,the relative expression levels of phosphorylated extracellular regulates protein kinase were 1.42±0.06,1.40±0.02,0.99±0 and 0.91±0.02,the relative expression levels of extracellular regulates protein kinase were 1.53±0.17,1.37±0.10,1.26±0.10 and 0.90±0.01,respectively.The above-mentioned indexes in experimental and combination groups were significantly different from those of blank control group(all P <0.05).Conclusion The activation of c-MET and its downstream signaling pathway is an important mechanism in gefitinib-resistant cells,so c-MET can be used as a potential therapeutic target to overcome acquired drug resistance in tongue squamous cell carcinoma.