食管癌相关基因4对弥漫性大B淋巴瘤细胞的作用及机制  

The effect and mechanism of esophageal cancer related gene 4 on diffuse large B-cell lymphoma

作　　者：杨林军[1] 汪正一 王建 张瑾[1] 金晓燕[1] Yang Linjun;Wang Zhengyi;Wang Jian;Zhang Jin;Jin Xiaoyan(Department of Surgical Oncology,Taizhou Municipal Hospital,surgical oncology department,Taizhou,318000,China)

机构地区：[1]浙江省台州市立医院肿瘤外科,台州318000

出　　处：《中华实验外科杂志》2024年第11期2538-2541,共4页Chinese Journal of Experimental Surgery

基　　金：浙江省科技厅公益技术研究计划项目(LGF20H280008);台州市科技计划项目(23ywb62、20ywa38)。

摘　　要：目的:探讨食管癌相关基因4(ECRG4)对弥漫性大B淋巴瘤(DLBCL)细胞增殖、凋亡和周期的影响。方法:制备食管癌相关基因4-绿色荧光蛋白(ECRG4-GFP)慢病毒,并将其体外感染ECRG4低表达的人弥漫大B淋巴瘤细胞WSU-DLCL2细胞株(WSU DLCL2)、人弥漫大B淋巴瘤细胞DHL4细胞株(DHL4)。通过免疫印迹法(Western blot)和实时荧光定量聚合酶链反应(RT-qPCR)检测ECRG4转染后,WSU DLCL2、DHL4细胞中ECRG4转录和表达水平;Hoechst 33342染色检测细胞核形改变;噻唑蓝(MTT)比色法检测细胞增殖活性;流式细胞仪观察细胞周期和凋亡水平。组间比较采用配对样本 t检验。 结果:Western blot和实时荧光定量RT-qPCR检测结果显示WSU DLCL2、DHL4两株淋巴瘤细胞经GFP-ECRG4慢病毒感染后,ECRG4转录和表达水平均高于对照组,(24.82±0.02比1.00±0.52, t=4.51, P<0.05);(34.54±0.03比1.00±0.06, t=4.99, P<0.05);Hoechst 33342染色结果证实ECRG4高表达后细胞核形改变;流式细胞仪检测结果显示ECRG4过表达的WSU DLCL2、DHL4细胞处于S期的细胞比例均高于未转染组[(50.60±4.30)%比(43.87±2.60)%, t=9.50, P<0.05;(48.33±2.15)%比(42.56±3.50)%, t=7.85, P<0.05];MTT结果证实ECRG4高表达后,WSU DLCL2、DHL4两株淋巴瘤细胞的增殖低于对照组,转染ECRG4 24 h[(32.00±0.06)%比(60.00±0.05)%, t=3.15, P<0.05;(56.00±0.03)%比(31.00±0.04)%, t=5.32, P<0.05];转染ECRG4 72 h后[(47.00±0.03)%比(110.00±0.03)%, t=6.15, P<0.05;(31.00±0.04)%比(100.00±0.01)%, t=9.11, P<0.05]。 结论:ECRG4基因可以通过阻滞细胞周期和诱导细胞凋亡的机制发挥抗DLBCL细胞增殖的作用。Objective To investigate the effect of esophageal cancer related gene 4(ECRG4)on the proliferation,apoptosis and cycle of diffuse large B-cell lymphoma(DLBCL).Methods The lentivirus ECRG4-green fluorescent protein(GFP)was prepared and infected in vitro to DLBCL cell lines WSU DLCL2 and DHL4 which low expressed ECRG4 protein.Western blotting and real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)were used to detect the transcription and expression level of ECRG4 in WSU DLCL2 and DHL4 cells transfected with ECRG4.Hoechst 33342 staining was used to detect the changes of nuclear shape;Cell proliferation activity was detected by methyl thiazolyl tetrazolium(MTT)colorimetric method.Cell cycle and apoptosis were observed by flow cytometry,Paired sample t-test was used for comparison between groups.ResultsWestern blotting and real-time fluorescence quantitative results showed that the transcription and expression levels of ECRG4 in WSU DLCL2 and DHL4 lymphoma cells infected with GFP-ECRG4 lentivirus were higher than those in the control group(24.82±0.02 vs.1.00±0.52.t=4.51,P<0.05);(34.54±0.03 vs.1.00±0.06,t=4.99,P<0.05);The results of Hoechst 33342 staining showed that the nuclear shape changed after ECRG4 high expression.Flow cytometry showed that the proportion of WSU DLCL2 and DHL4 cells with ECRG4 overexpression in S phase was higher than that in the non-transfected group[(50.60±4.30)%vs.(43.87±2.60)%,t=9.50,P<0.05;(48.33±2.15)%vs.(42.56±3.50)%,t=7.85,P<0.05];MTT results showed that the proliferation of WSU DLCL2 and DHL4 lymphoma cells after ECRG4 overexpression was lower than that of the control group.24 h after ECRG4 transfection[(32.00±0.06)%vs.(60.00±0.05)%,t=3.15,P<0.05;(56.00±0.03)%vs.(31.00±0.04)%,t=5.32,P<0.05];After transfection with ECRG4 for 72 h,the ratio of[(47.00±0.03)%vs.(110.00±0.03)%,t=6.15,P<0.05;(31.00±0.04)%vs.(100.00±0.01)%,t=9.11,P<0.05].Conclusion ECRG4 gene can inhibit the proliferation of diffuse large B lymphoma cells by blocking cell cycle and in

关 键 词：食管癌相关基因4 弥漫性大B淋巴瘤细胞 细胞增殖 细胞凋亡 

分 类 号：R735.1[医药卫生—肿瘤] R733.1[医药卫生—临床医学]