摘要
目的探讨肾母细胞瘤基因1(WT1)对鼻咽癌细胞(CNE)-2表皮生长因子受体(EGFR)的靶向作用,证实WT1对CNE-2免疫活性的影响。方法采用WT1对CNE-2进行处理,借助蛋白质免疫印迹法(Western blot)对EGFR的蛋白表达情况进行检测,借助CKK-8观察CNE-2增殖能力情况,借助流式细胞术观察CNE-2凋亡情况,借助基质胶侵袭试验(Transwell)测定CNE-2侵袭情况。结果 Western blot测定结果显示,采用WT1组信号强度明显低于二甲亚砜(DMSO)组和空白组,差异均有统计学意义(P<0.05);WT1组检测灰度值为0.358±0.148,DMSO组为1.370±0.068,空白组为1.390±0.077,WT1组检测灰度值明显低于DMSO组和空白组,差异均有统计学意义(P<0.05)。CKK-8检测结果显示,采用WT1进行处理后3、6、9、12h,抑制值表现为逐渐增大,差异有统计学意义(P<0.05)。流式细胞术检测结果显示,空白组细胞凋亡率为(0.832±0.207)%,DMSO组为(2.081±1.221)%,WT1组为(20.731±1.104)%,WT1组细胞凋亡率高于空白组和DMSO组,差异均有统计学意义(P<0.05)。Transwell测定结果显示,DMSO组穿透细胞数量为67.21±2.17,WT1组为20.13±2.25,WT1组穿透细胞数量低于DMSO组,差异有统计学意义(P<0.05)。结论 WT1抗原能够作为CNE-2EGFR的靶向基因,可较好地实现对CNE-2EGFR免疫活性的抑制。
Objective To investigate the targeting effect of nephroblastoma gene 1(WT1)on epidermal growth factor receptor(EGFR)of nasopharyngeal carcinoma CNE-2 cells,and to confirm the effect of WT1 on immune activity of CNE-2 cells.Methods The cells were treated by WT1,the protein expression of EGFR was detected by Western blot,the proliferation capacity of CNE-2 cells was observed by CKK-8,the apoptosis of CNE-2 cells was observed by flow cytometry,and the invasion of CNE-2 cells was determined by Transwell assay.Results Western blot showed that the signal intensity of WT1 group was significantly lower than that of dimethyl sulfoxide(DMSO)group and control group(P<0.05).The gray value detected in the WT1 group was(0.358±0.148),which in the DMSO group was(1.370±0.068),and that of the blank group was(1.390±0.077).The gray value in the WT1 group was significantly lower than that in the DMSO group and the blank group,and the difference was statistically significant(P<0.05).CKK-8 test results showed that the inhibition value of WT1 increased gradually after 3,6,9 and 12 hof treatment,and the difference was statistically significant(P<0.05).The results of flow cytometry showed that the apoptosis rate of the blank group was(0.832±0.207)%,which in the DMSO group was(2.081±1.221)%,and that of the WT1 group was(20.731±1.104)%.The apoptosis rate of the WT1 group was higher than that of the DMSO group,and the difference was statistically significant(P<0.05).Transwell assay showed that the number of penetrating cells in DMSO group was 67.21±2.17,in WT1 group was 20.13±2.25,and the number of penetrating cells in WT1 group was lower than that in DMSO group,and the difference was statistically significant(P<0.05).Conclusion WT1 antigen can be used as a target gene of EGFR in CNE-2 cells,and can better inhibit the EGFR immune activity of CNE-2 cells.
作者
方慧云
程伟民
季明芳
俞霞
李付贵
何洁冰
FANG Huiyun;CHENG Weimin;JI Mingfang;YU Xia;LI Fugui;HE Jiebing(Tumor Research Institute,Zhongshan Hospital Affiliated to Sun Yat-Sen University,Zhongshan,Guangdong528403,China)
出处
《检验医学与临床》
CAS
2019年第7期878-881,共4页
Laboratory Medicine and Clinic
基金
2014年广东省中山市科技计划立项课题资助项目(2014A1FC071)
关键词
鼻咽癌
肾母细胞瘤基因1
靶向作用
免疫活性
表皮生长因子受体
nasopharyngeal carcinoma
targeting effect of nephroblastoma gene 1
targeted action
immune activity
epidermal growth factor receptor
