蛋白酶体抑制剂硼替佐米对食管鳞状细胞癌生长的影响

Effect of proteasome inhibitor bortezomib on the growth of esophageal squamous cell carcinoma

目的探讨蛋白酶体抑制剂硼替佐米对食管鳞状细胞癌生长的影响。方法基于全基因组测序数据,利用药物靶标数据库DGIdb筛选食管鳞状细胞癌相关药物靶基因,采用DAVID软件进行KEGG信号通路富集分析。采用MTT法检测硼替佐米对KYSE30、KYSE180、KYSE150、TE1、KYSE510等5株食管鳞状细胞癌细胞生长的影响,以DMSO作用为相应对照组。裸鼠体外成瘤后分别腹腔注射生理盐水(对照组)及硼替佐米(硼替佐米组),观察硼替佐米对裸鼠移植瘤体积及重量的影响;免疫组织化学法检测裸鼠移植瘤细胞Ki-67蛋白的表达。结果469例食管鳞状细胞癌的基因组学测序数据在DGIdb数据库中共鉴定出307个药物靶基因,突变频率>2.5%的显著突变药物靶基因包括PIK3CA、NOTCH1、CDKN2A、ERBB4等;药物靶基因富集的信号通路有RTK-RAS、PI3K/AKT/mTOR、NOTCH、ERBB信号通路、细胞周期和蛋白酶体途径等。MTT实验结果显示,硼替佐米处理后,5株食管鳞状细胞癌细胞的增殖能力均较对照组降低(P<0.05);与对照组相比,硼替佐米组裸鼠移植瘤体积减小[(1909.18±533.40)mm3 vs(1065.83±283.94)mm3,P=0.007],移植瘤重量降低[(1.60±0.36)g vs(0.98±0.30)g,P=0.009]。免疫组织化学法检测结果显示,与对照组比较,硼替佐米组裸鼠移植瘤组织中Ki-67表达降低(86.32±4.51 vs 43.83±3.22,P=0.001)。结论蛋白酶体抑制剂硼替佐米在体外和体内均可抑制食管鳞状细胞癌细胞生长。

Objective To investigate the effect of proteasome inhibitor bortezomib on the growth of esophageal squamous cell carcinoma(ESCC).Methods Based on whole genome sequencing data,DGIdb database was used to screen esophageal squamous cell carcinoma-related druggable genes,and KEGG signal pathway enrichment analysis was performed using DAVID software.The effect of bortezomib on the growth of 5 ESCC cells including KYSE30,KYSE180,KYSE150,TE1and KYSE510 were measured by MTT,and DMSO-treated cells were used as the corresponding control group.Nude mice were injected intraperitoneally with saline(control group)and bortezomib(bortezomib group)after tumor formation in vitro.The effects of bortezomib on the volume and weight of xenografts in nude mice were observed.Immunohisto-chemistry was performed to determine the expression of Ki-67 protein in xenograft tumor tissues treated with bortezomib.Results A total of 307 druggable genes were identified from genomics sequencing data of 469 ESCC cases in the DGIdb database,and the significantly mutated druggable genes with mutation frequencies>2.5%included PIK3CA,NOTCH1,CDKN2A,ERBB4,etc.The enriched signaling pathways of all druggable genes included RTK-RAS,PI3K/AKT/mTOR,NOTCH,ERBB signaling pathway,cell cycle and proteasome pathway,etc.MTT results showed that the proliferation of 5 ESCC cell lines treated with bortezomib were significantly inhibited compared with control group(P<0.05).Compared with the control group,the tumor volume of the bortezomib group was significantly decreased[(1909.18±533.40)mm3 vs(1065.83±283.94)mm3,P<0.01],and the tumor weight of the bortezomib group was also significantly decreased[(1.60±0.36)g vs(0.98±0.30)g,P<0.01].Immunohistochemistry showed that the expression of Ki-67 in the bortezomib group was significantly decreased compared with the control group(43.83±3.22 vs 86.32±4.51,P<0.01).Conclusion Bortezomib can inhibit the cell growth of ESCC in vitro and in vivo.