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目的:探讨葛根素对Lewis肺癌细胞中表皮生长因子受体酪氨酸激酶(EGFR-TK)特异性抑制机理。方法:不同浓度葛根素组加入到EGFR-TK、Lewis肺癌细胞株中,利用ELISA法研究葛根素对EGFR-TK的抑制作用机理,利用MTT法测定葛根素对Lewis肺癌细胞株生长的抑制作用,利用CD光谱法监测葛根素对EGFR-TK二级结构的影响,流式细胞术检测细胞凋亡率,采用Transwell小室法检测细胞侵袭率。结果:葛根素是可逆的竞争性EGFR-TK抑制剂(半抑制率IC50和抑制常数Ki分别为4.18±0.34μg/ml和2.12±0.21μg/ml),此外葛根素对Lewis肺癌细胞生长和侵袭具有良好的抑制能力,并能够明显的诱导Lewis肺癌细胞的凋亡;CD光谱分析表明葛根素诱导EGFR-TK的构象发生部分改变,α-螺旋含量逐渐增加,无规则卷曲逐渐减少。结论:葛根素能够特异性的抑制EGFR-TK的活性,表现出来良好的抗Lewis肺癌细胞生长、侵袭和诱导细胞凋亡的药理活性。
AIM: To investigate the specific inhibition mechanism of puerarin on epidermal growth factor receptor tyrosine kinase (EGFR-TK) in Lewis lung carcinoma cells. Methods: The puerarin groups were added into EGFR-TK and Lewis lung cancer cell lines with different concentrations. The inhibitory mechanism of puerarin on EGFR-TK was studied by ELISA. The inhibitory effect of puerarin on the growth of Lewis lung cancer cell lines was determined by MTT assay. The effect of puerarin on the secondary structure of EGFR-TK was monitored by CD spectroscopy. The apoptosis rate was detected by flow cytometry. The cell invasion rate was detected by Transwell chamber assay. RESULTS: Puerarin was a reversible competitive inhibitor of EGFR-TK (IC50 and IC50 of 4.18 ± 0.34μg / ml and 2.12 ± 0.21μg / ml, respectively). In addition, puerarin inhibited the growth and invasion of Lewis lung carcinoma cells The results showed that the conformation of EGFR-TK induced by puerarin changed partly, the α-helix content increased gradually, and the irregular curl decreased gradually. CONCLUSION: Puerarin can specifically inhibit the activity of EGFR-TK and show good pharmacological activity against Lewis lung cancer cells in growth, invasion and apoptosis induction.