肝细胞生长因子诱导慢性髓细胞性白血病K562细胞抗凋亡效应研究

发布时间：2018-04-24 10:33

本文选题：肝细胞生长因子 + 慢性髓细胞性白血病　；参考：《中国药学杂志》2017年03期

【摘要】：目的观察肝细胞生长因子(hepatocyte growth factor,HGF)对经凋亡诱导剂足叶乙苷(etoposide,VP-16)诱导后慢性髓细胞性白血病(CML)K562细胞凋亡的抑制作用,并分析其分子机制。方法采用苏木素-伊红(HE)染色、吖啶橙染色(AO)染色对凋亡细胞形态特征变化进行定性/半定量分析;采用Annexin V-FITC/PI双染、JC-1染色检测细胞膜表面的PS外翻和完整性及线粒体膜电位分析凋亡细胞生化特征变化;采用荧光定量聚合酶链反应(PCR)检测Bcl-2、Bax、Caspase-3、Caspase-9等凋亡相关基因mRNA表达的变化,综合评价HGF抗凋亡效应,并阐述其分子机制。结果 HE法、AO法发现HGF+VP-16组凋亡率明显低于VP-16组(P0.05、P0.05),提示HGF可显著抑制凋亡的发生;Annexin V-FITC/PI双染法、JC-1染色法发现HGF+VP-16组早期凋亡细胞明显低于VP-16组(P0.05、P0.001),提示HGF具有抗K562细胞早期凋亡效应;凋亡相关基因mRNA表达检测结果发现,HGF+VP-16组的Bcl-2 mRNA表达量明显高于VP-16组(P0.001),而Bax mRNA、Caspase-3 mRNA、Caspase-9 mRNA表达量明显低于VP-16组(P0.05、P0.001、P0.001),证实HGF抑制凋亡基因表达,同时促进抗凋亡基因的表达,提示HGF具抗凋亡效应。结论 HGF显著抑制经VP-16诱导的CML K562细胞的凋亡,该抗凋亡效应可能通过HGF/c-Met途径调控PI3K/AKT通路而实现。
[Abstract]:Objective to observe the inhibitory effect of hepatocyte growth factor (HGF) on the apoptosis of chronic myeloid leukemia (CML) K562 cells induced by etoposideside VP-16, and to analyze its molecular mechanism. Methods the morphological changes of apoptotic cells were qualitatively and semi-quantitatively analyzed by hematoxylin eosin (HE) staining and acridine orange staining (AOO). Annexin V-FITC/PI double staining JC-1 staining was used to detect PS valgus and integrity on cell membrane surface and mitochondrial membrane potential to analyze the biochemical characteristics of apoptotic cells, and fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the changes of mRNA expression of apoptosis-related genes such as Bcl-2Caspase-3Caspase-9. The antiapoptotic effect of HGF was comprehensively evaluated and its molecular mechanism was expounded. Results the apoptosis rate of HGF VP-16 group was significantly lower than that of VP-16 group P0.05 P0.05A, which suggested that HGF could significantly inhibit apoptosis. The results showed that the early apoptotic cells in HGF VP-16 group were significantly lower than that in VP-16 group P0.05 and P0.001, suggesting that HGF had anti-K562 thin cells. Early apoptosis; The expression of Bcl-2 mRNA in HGF VP-16 group was significantly higher than that in VP-16 group, while the expression of Caspase-3 mRNA-caspase-9 mRNA in Bax mRNAs was significantly lower than that in VP-16 group. It was confirmed that HGF inhibited the expression of apoptotic gene and promoted the expression of anti-apoptotic gene. The results suggest that HGF has anti-apoptosis effect. Conclusion HGF significantly inhibits the apoptosis of CML K562 cells induced by VP-16, and the anti-apoptotic effect may be achieved through the regulation of PI3K/AKT pathway by HGF/c-Met pathway.
【作者单位】：宁波市第一医院;温州医科大学浙江省医学遗传学重点实验室;宁波市泌尿肾病医院;
【基金】：浙江省医药卫生科技项目(2014KYB243) 宁波市科技计划( 2014C50015、2013A610244、2013A610219、2010A610031)
【分类号】：R73-36

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