肺癌体外细胞药敏试验指导盐酸埃克替尼临床用药的基础研究

发布时间：2018-05-14 11:39

本文选题：埃克替尼 + 肺癌　；参考：《川北医学院》2017年硕士论文

【摘要】：目的:初步探讨盐酸埃克替尼对肺癌细胞的增殖抑制作用与EGFR基因突变状态的关系,为临床上细胞药敏试验作为EGFR-TKIs疗效预测方法之一提供参考。方法:以体外培养的A549和HCC827肺癌细胞株为实验对象,通过形态学观察和免疫细胞化学染色法鉴定两细胞株,采用x TAG液相芯片技术检测细胞株EGFR基因突变状态,CCK-8法检测埃克替尼对两细胞株的增殖抑制作用,Annexin V-FITC/PI双染法检测埃克替尼对两细胞株的促凋亡作用,实验数据应用SPSS21.0软件进行统计学分析,检验水准为P0.05。结果:1)两细胞株形态学表现和免疫细胞化学染色情况均符合A549和HCC827肺癌细胞株的特性。2)EGFR基因突变状态:A549细胞株为野生型,HCC827细胞株为19外显子缺失突变型(p.E746_A750del)。3)埃克替尼对A549细胞株作用48h的半数抑制浓度(IC50)为9.65±0.78μmol/L,对HCC827细胞株作用的IC50为0.12±0.01μmol/L,两者比较差异有统计学意义(P=0.000)。4)选用0.1μmol/L盐酸埃克替尼作用A549和HCC827细胞株48h,两者细胞抑制率分别为0和(45.31±1.33)%(P=0.000),细胞凋亡率分别为(1.81±0.47)%和(62.25±2.45)%(P=0.000);选用1μmol/L盐酸埃克替尼作用A549和HCC827细胞株48h,两者细胞抑制率分别为(18.95±0.78)%和(75.53±2.98)%(P=0.000),两者细胞凋亡率分别为(39.91±2.04)%和(89.26±3.81)%(P=0.000)。结论:1)本课题初步证实了免疫细胞化学染色鉴定A549和HCC827肺癌细胞株的可行性,研究中的免疫细胞化学染色结果可能成为其他研究中鉴定该两细胞株时的参考依据;2)课题中埃克替尼对EGFR基因突变型的HCC827肺腺癌细胞株的增殖抑制作用明显强于EGFR基因野生型的A549肺癌细胞株,提示研究方法真实、可靠,可为临床上EGFR-TKIs细胞药敏试验的实施提供一定参考。
[Abstract]:Objective: to investigate the relationship between the inhibitory effect of Ektinib hydrochloride on the proliferation of lung cancer cells and the mutation status of EGFR gene, and to provide a reference for clinical chemosensitivity test as one of the methods for predicting the curative effect of EGFR-TKIs. Methods: A549 and HCC827 lung cancer cell lines were cultured in vitro and identified by morphological observation and immunocytochemical staining. X TAG liquid chip technique was used to detect the mutation status of EGFR gene in cell line and CCK-8 method was used to detect the inhibitory effect of Ectini on the proliferation of two cell lines. Annexin V-FITC/PI double staining was used to detect the apoptosis-promoting effect of Ectini on the two cell lines. The experimental data were analyzed by SPSS21.0 software, and the test level was P0.05. Results the morphological features and immunocytochemical staining of the two cell lines were consistent with the characteristics of A549 and HCC827 lung cancer cell lines. IC50 was 9.65 卤0.78 渭 mol / L in A549 cell line and 0.12 卤0.01 渭 mol / L in HCC827 cell line for 48 h. The difference was statistically significant (P < 0.05). The inhibition rate of A549 cell line and HCC827 cell line treated with 0.1 渭 mol/L epetinib hydrochloride for 48 h was significant. The cell apoptosis rates were 1.81 卤0.47% and 62.25 卤2.45%, respectively, and the inhibitory rates of 1 渭 mol/L hydrochloride on A549 and HCC827 cells for 48 h were 18.95 卤0.78% and 75.53 卤2.98%, respectively, and the apoptotic rates were 39.91 卤2.04% and 89.26 卤3.81%, respectively. Conclusion 1) the feasibility of identification of A549 and HCC827 lung cancer cell lines by immunocytochemical staining was preliminarily confirmed. The results of immunocytochemical staining in the study may be the reference basis for identifying the two cell lines in other studies. The inhibitory effect of Ectini on the proliferation of EGFR mutant HCC827 lung adenocarcinoma cell lines is significantly stronger than that of other studies. EGFR gene wild-type lung cancer cell line A549, The results suggest that the research method is true and reliable, and can provide some reference for the clinical application of EGFR-TKIs cell drug sensitivity test.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2

【参考文献】