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目的:观察黄芩提取化合物单体成分SBX对人NSCLC细胞系H1975和H1944细胞株增殖的影响,初步探讨SBX对NSCLC细胞系抑制的机制。方法:体外培养人H1975和H1944细胞株,以0、10、25、50、100和200μmol/L的SBX处理细胞,应用MTT法检测SBX对细胞增殖的影响,筛选出敏感细胞株。应用流式细胞术检测SBX对敏感细胞株细胞周期的影响,应用Protein Pathway Array技术检测SBX对敏感细胞株蛋白表达的影响。结果:SBX对两种类型的NSCLC细胞株均有抑制作用,并呈剂量依赖性,对H1975和H1944细胞株的IC50值分别为(38.3±1.25)和(62.5±1.76)μmol/L。SBX对H1975和H1944细胞抑制作用比较,对H1975细胞抑制作用更强,P<0.001。流式细胞术显示,SBX对H1975细胞周期的影响主要是引起G0/G1期阻滞和凋亡增加,与空白对照组相比,P<0.001。50和100μg/mL的SBX分别作用于H1975细胞后,阻滞在G0/G1期的细胞由35.47%分别升至54.36%和73.37%,S期的细胞由61.60%分别降至41.63%和19.14%。应用Protein Pathway Array技术分析SBX对H1975蛋白表达的影响显示,共检测166个磷酸化和非磷酸化蛋白,其中53个蛋白表达,有差异性表达的蛋白共15个。其中Cdk6、EGFR、Survivin和mTor蛋白表达水平上调,而p27、p-RB、Cyclin D1、Cyclin E、XIAP、p53、p-p53、p-AKT、Akt、Notch4和Cdk4蛋白表达水平下调。结论:SBX能够抑制人NSCLC细胞系H1975和H1944细胞株的增殖,敏感细胞株为H1975;SBX主要影响H1975的EGFR和Notch信号传导通路,并通过以上信号传导通路影响细胞周期相关蛋白表达水平的变化,导致细胞周期G0/G1期阻滞,凋亡增加。
OBJECTIVE: To observe the effects of SBX, an extract of Scutellaria baicalensis Georgi, on the proliferation of human NSCLC cell lines H1975 and H1944, and to explore the mechanism of SBX inhibition on NSCLC cell lines. METHODS: Human H1975 and H1944 cell lines were cultured in vitro. Cells were treated with 0, 10, 25, 50, 100 and 200 μmol / L SBX. The effect of SBX on cell proliferation was detected by MTT assay. The effect of SBX on the cell cycle of sensitive cell lines was detected by flow cytometry. Protein Pathway Array was used to detect the effect of SBX on the protein expression of the sensitive cell lines. RESULTS: SBX inhibited both NSCLC cell lines in a dose-dependent manner, with IC50 values of (38.3 ± 1.25) and (62.5 ± 1.76) μmol / L for H1975 and H1944 cell lines, respectively. Compared with H1975 and H1944 cells, SBX showed stronger inhibitory effect on H1975 cells (P <0.001). Flow cytometry showed that the effect of SBX on the cell cycle of H1975 was mainly caused by the arrest of G0 / G1 phase and the increase of apoptosis. Compared with the blank control group, P <0.001.50 and 100 μg / mL of SBX acted on H1975 cells Afterwards, the number of cells arrested in G0 / G1 phase increased from 35.47% to 54.36% and 73.37%, respectively, and the S phase cells decreased from 61.60% to 41.63% and 19.14%, respectively. Analysis of the effect of SBX on the expression of H1975 protein using Protein Pathway Array showed that a total of 166 phosphorylated and non-phosphorylated proteins were detected, of which 53 proteins were expressed and 15 proteins were differentially expressed. The expression of Cdk6, EGFR, Survivin and mTor were up-regulated while the expressions of p27, p-RB, Cyclin D1, Cyclin E, XIAP, p53, p-AKT, Akt, Notch4 and Cdk4 were down-regulated. CONCLUSIONS: SBX can inhibit the proliferation of H1975 and H1944 cell lines, and the sensitive cell line is H1975. SBX mainly affects the EGFR and Notch signaling pathways of H1975 and affects the expression of cell cycle-related proteins through the above signal transduction pathways , Leading to cell cycle G0 / G1 arrest, apoptosis increased.