摘要
目的探讨表皮生长因子受体(EGFR)基因介导酪氨酸激酶(JAK)/信号转导与转录激活因子(STAT)信号通路对胶质瘤细胞凋亡的调控机制。方法收集甘肃省人民医院2017年1月至2019年6月胶质瘤患者手术切除的胶质瘤组织及瘤旁组织。细胞进行细胞分组与转染,分成7组:Blank组、NC组、oe-EGFR组、小干扰RNA(siRNA)-EGFR组、Gefitinib组、NSC 228155组和siRNA-EGFR+NSC 228155组。实时定量反转录聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)法检测胶质瘤组织和转染后各组细胞中EGFR、JAK1、STAT3、B细胞淋巴瘤/白血病-2(bcl-2)和bcl-2相关X蛋白(bax) mRNA和蛋白的表达。噻唑蓝(MTT)法和流式细胞术分别检测各组细胞转染后细胞增殖与凋亡。两组间比较采用Student’st检验,多组间比较应用单因素方差分析。结果胶质瘤组织中的EGFR、JAK1、STAT3、bcl-2 mRNA(EGFR:t=13.630,P<0.01;JAK1:t=16.650,P<0.01;STAT3:t=17.420,P<0.01;bcl-2:t=17.010,P<0.01)和蛋白(EGFR:t=15.590,P<0.01;JAK1:t=11.680,P<0.01;STAT3:t=9.295,P<0.01;bcl-2:t=13.960,P<0.01)表达量显著高于癌旁组织,而Bax mRNA(t=21.480,P<0.01)和蛋白(t=10.950,P<0.01)表达量显著低于癌旁组织,差异有统计学意义(均P<0.05)。Blank组和NC组组间差异无统计学意义(均P>0.05);siRNA-EGFR组和Gefitinib组Bax mRNA(F=48.300,P<0.01)和蛋白(F=28.330,P<0.01)表达量显著高于NC组,而EGFR、JAK1、STAT3、bcl-2 mRNA(EGFR:F=138.000,P<0.01;JAK1:F=234.300,P<0.01;STAT3:F=106.300,P<0.01;bcl-2:F=186.500,P<0.01)和蛋白(EGFR:F=63.540,P<0.01;JAK1:F=61.720,P<0.01;STAT3:F=54.660,P<0.01;bcl-2:F=77.480,P<0.01)表达量显著低于NC组,细胞增殖能力明显低于NC组(24 h:F=23.650,P<0.01;48 h:F=44.450,P<0.01;72 h:F=32.160,P<0.01),而细胞凋亡明显高于NC组(F=50.810,P<0.01),差异有统计学意义(均P<0.05);oe-EGFR组和NSC 228155组各因子和指标趋势逆转,差异有统计学意义(均P<0.05);而siRNA-EGFR+NSC 228155组各因子和指标与NC组相比差异无统计学意�
Objective To explore the regulatory mechanism of janus kinase(JAK)/signal transducer and activators of transcription(STAT)signaling pathway mediated by epidermal growth factor receptor(EGFR)gene on glioma cell apoptosis.Methods The glioma tissue and the adjacent tissue were collected.Cells were divided into 7 groups:blank group,normal control group,oe-EGFR group,siRNA-EGFR group,Gefitinib group,NSC 228155 group and small interfering RNA(siRNA)-EGFR+NSC 228155 group.The mRNA and protein expression of EGFR,JAK1,STAT3,B cell lymphoma/leukemia-2(bcl-2)and bcl-2 associated X protein(bax)mRNA in tissues was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and Western blotting respectively.The methyl thiazolyl tetrazolium(MTT)method and flow cytometry were used to detect the cell proliferation and apoptosis after transfection.Student′s t test was used for comparison between groups,and one-way ANOVA was used for comparison among groups.Results The expression of EGFR,JAK1,STAT3 and bcl-2 mRNA(EGFR:t=13.630,P<0.01;JAK1:t=16.650,P<0.01;STAT3:t=17.420,P<0.01;bcl-2:t=17.010,P<0.01)and protein(EGFR:t=15.590,P<0.01;JAK1:t=11.680,P<0.01;STAT3:t=9.295,P<0.01;bcl-2:t=13.960,P<0.01)in gliomas increased significantly,while Bax mRNA(t=21.480,P<0.01)and protein(t=10.950,P<0.01)decreased significantly,with statistical differences(all P<0.05).Compared with blank group and normal control group,the expression of Bax mRNA(F=48.300,P<0.01)and protein(F=28.330,P<0.01)in siRNA-EGFR group and Gefitinib group increased significantly,while the expression of EGFR,JAK1,STAT3 and bcl-2 mRNA(EGFR:F=138.000,P<0.01;JAK1:F=234.300,P<0.01;STAT3:F=106.300,P<0.01;bcl-2:F=186.500,P<0.01)and protein(EGFR:F=63.540,P<0.01;JAK1:F=61.720,P<0.01;STAT3:F=54.660,P<0.01;bcl-2:F=77.480,P<0.01)decreased significantly,cell proliferation ability decreased(24 h:F=23.650,P<0.01;48 h:F=44.450,P<0.01;72 h:F=32.160,P<0.01),and apoptosis increased(F=50.810,P<0.01),with statistical differences(all P<0.05);the opposite trends were fo
作者
焦克平
包丽娟
陈恒三
赵玉梅
Jiao Keping;Bao Lijuan;Chen Hengsan;Zhao Yumei(Department of Neurosurgery,Gansu Provincial Hospital,Lanzhou 731200,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第9期1654-1658,共5页
Chinese Journal of Experimental Surgery
关键词
表皮生长因子受体基因
酪氨酸激酶/信号转导与转录激活因子信号通路
胶质瘤
增殖
凋亡
Epidermal growth factor receptor gene
Janus kinase/signal transducer and activators of transcription signaling pathway
Glioma
Proliferation
Apoptosis
