Hsp90抑制剂Ganetespib联合培美曲塞对肺腺癌细胞的影响  

Effect of Hsp90 Inhibitor Ganetespib Combined with Pemetrexed on Lung Adenocarcinoma Cells

作　　者：杨晟强 王平[1] YANG Shengqiang;WANG Ping(Dept.of Thoracic Surgery,The 2nd Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650032,China)

机构地区：[1]昆明医科大学第二附属医院胸外科,云南昆明650032

出　　处：《昆明医科大学学报》2024年第7期30-41,共12页Journal of Kunming Medical University

基　　金：云南省科技厅科技计划基金资助项目(202201AY070001-120)。

摘　　要：目的探索Ganetespib和培美曲塞体外对人肺腺癌细胞增殖、凋亡和相关信号通路的影响。方法人肺腺癌细胞HCC827、H1975、A549分组(Ganetespib组、培美曲塞组、2药联用组),同时设置对照组(DMSO 0.2%),培养48 h后,Cell Titer-Glo(CTG)法体外检测抑制细胞增殖能力,Western blot检测细胞中Akt、p-Akt、Bcl-2蛋白表达水平,细胞凋亡实验检测细胞凋亡率,RNA-seq检测2药物对于HCC827细胞转录组水平的影响。结果抑制细胞增殖效果Ganetespib>培美曲塞(P<0.001),2药联用时抑制效果较单药增强(P<0.0001);细胞凋亡率Ganetespib>培美曲塞(P<0.05),2药联用时HCC827细胞凋亡率较单药升高;Ganetespib组Akt蛋白磷酸化水平降低(P<0.05),培美曲塞组Akt蛋白磷酸化水平升高(P<0.05),2药联用组结果与Ganetespib组无明显差别,Bcl-2蛋白仅在A549细胞中下调(P<0.01);2药物对HCC827细胞株的基因转录水平有影响,Ganetespib组上调的差异表达基因为102个,下调有27个,培美曲塞组的上调差异表达基因为53个,下调有8个。结论Ganetespib联用培美曲塞可明显抑制肺腺癌细胞HCC827、H1975和A549的增殖,诱导细胞凋亡,其分子机制可能是通过PI3K/AKT信号通路的蛋白质磷酸化来抑制肺腺癌细胞增殖。联合用药可一定程度增强抑制肺腺癌细胞增殖作用,具体机制需进一步研究。Objective To investigate the effects of Ganetespib and Pemetrexed on the proliferation,apoptosis and related signaling pathways of human lung adenocarcinoma cells in vitro.Methods Human lung adenocarcinoma cells HCC827,H1975,and A549 were divided into groups(Ganetespib group,pemetrexed group,combined treatment group),and a control group(DMSO 0.2%)was set up.After culturing for 48 hours,the Cell Titer-Glo(CTG)method was used to detect the inhibitory effects of cell proliferation in vitro,and Western blot was used to detect the expression levels of Akt,p-Akt,and Bcl-2 proteins in cells.The apoptosis experiment was used to detect the apoptosis rate,and RNA-seq was used to detect the effects of the two drugs on the transcription level of the HCC827 cell line.Results The inhibitory effects of cell proliferation were as follows:Ganetespib>pemetrexed(P<0.001),and the inhibitory effect was enhanced when the two drugs were combined(P<0.0001);the apoptosis rate was as follows:Ganetespib>pemetrexed(P<0.05),and the apoptosis rate of HCC827 cells increased when the two drugs were combined;the phosphorylation level of Akt protein was lower in the Ganetespib group(P<0.05),and the phosphorylation level of Akt protein was higher in the pemetrexed group(P<0.05),and the results of the combined treatment group were similar to those of the Ganetespib group,and Bcl-2 protein was only downregulated in A549 cells(P<0.01);the two drugs had an effect on the gene transcription level of the HCC827 cell line,with 102 upregulated differentially expressed genes and 27 downregulated genes in the Ganetespib group,53 upregulated differentially expressed genes and 8 downregulated genes in the pemetrexed group.Conclusion The combination of Ganetespib and pemetrexed can significantly inhibit the proliferation of lung adenocarcinoma cells HCC827,H1975,and A549,induce cell apoptosis.The molecular mechanism may be through the inhibition of lung adenocarcinoma cell proliferation by protein phosphorylation in the PI3K/AKT signaling pathway.The transcript

关 键 词：肺腺癌 Ganetespib 培美曲塞 

分 类 号：R734.2[医药卫生—肿瘤]