首页|MYD88过表达对弥漫大B细胞淋巴瘤细胞增殖、凋亡的影响及其作用机制

MYD88过表达对弥漫大B细胞淋巴瘤细胞增殖、凋亡的影响及其作用机制

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目的 探讨MYD88基因过表达对人弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)细胞增殖、凋亡的影响及其作用机制.方法 采用质粒转染法将过表达MYD88 L265P基因的pEGFP-C2-MYD88转染DLBCL细胞;实验分为空白对照组、阴性对照组和MYD88 L265P过表达组.倒置荧光显微镜下观察MYD88 L265P过表达后荧光表达;运用RT-PCR、West-ern blot法检测MYD88 L265P过表达前后DLBCL细胞的MYD88 L265P、IRAK4、NF-κB和BCL2的mRNA及蛋白表达水平;应用CCK-8法检测DLBCL细胞增殖;采用Hoechst染色法检测DLBCL细胞凋亡情况.结果 MYD88 L265P过表达后,与空白对照组(0.670 4±0.017 5)和阴性对照组(0.715 3±0.019 6)相比,MYD88 L265P过表达组(1.157 2±0.010 2)的增殖率显著增高,均有统计学意义(P均<0.05).MYD88 L265P过表达后,与空白对照组(0.69±0.04)和阴性对照组(0.81±0.07)相比,MYD88 L265P过表达组(0.48±0.05)的凋亡率明显降低,均有统计学意义(P均<0.05).MYD88 L265P过表达后,与空白对照组(mRNA:1.015 8±0.011 5、0.987 3±0.010 2、1.007 6±0.015 3;蛋白:0.183 4±0.058 9、0.096 8±0.015 7、0.147 5 ±0.041 8)和阴性对照组(mRNA:0.913 2±0.009 8、1.003 2±0.015 6、0.932 7±0.011 2;蛋白:0.187 9±0.042 3、0.088 9± 0.051 3、0.134 8±0.050 1)相比,MYD88 L265P过表达组IRAK4、NF-κB和抗凋亡蛋白BCL2的mRNA(3.243 2±0.013 6、2.976 6±0.021 3、1.585 9±0.019 8)及蛋白表达(0.452 7±0.052 4、0.218 9±0.047 5、0.301 4±0.059 8)明显增高,均有统计意义(P均<0.05).结论 MYD88 L265P过表达后,DLBCL细胞的凋亡率下降,细胞增殖率上升,其机制可能与MYD88 L265P基因突变激活和放大了NF-κB通路,促进抗凋亡蛋白BCL2的过表达有关.
Effects of MYD88 overexpression on proliferation and apoptosis of diffuse large B cell lymphoma cells and its mechanism
Purpose To investigate the effect of MYD88 gene overexpression on the proliferation and apoptosis of human diffuse large B cell lymphoma(DLBCL)cells,and to prelimi-narily explore the mechanism of MYD88 gene action.Methods PEGFP-C2-MYD88 overexpressing MYD88 L265P gene was transfected into DLBCL cells by plasmid transfection.The exper-iment was divided into blank control group,negative control group and MYD88 L265P overexpression group.The fluores-cence expression of MYD88 L265P after overexpression was ob-served under inverted fluorescence microscope.RT-PCR and Western blot were used to detect the mRNA and protein expres-sion of MYD88 L265P,IRAK4,NF-κB and BCL2 in DLBCL cells before and after overexpression of MYD88 L265.CCK8 method was used to detect DLBCL cells proliferation and Ho-echst staining was used to detect DLBCL cells apoptosis.Re-sults After overexpression of MYD88 L265P,compared with the blank control group(0.670 4±0.017 5)and the negative control group(0.715 3±0.019 6),the MYD88L265P overex-pression group(1.157 2±0.010 2)increased significantly,with statistical significance(all P<0.05).After overexpression of MYD88 L265P,compared with the blank control group(0.69 ±0.04)and the negative control group(0.81±0.07),the MYD88L265P overexpression group(0.48±0.05)was signifi-cantly decreased,with statistical significance(all P<0.05).After overexpression of MYD88 L265P,compared with the blank control group(mRNA:1.0158±0.0115,0.987 3±0.010 2,1.007 6±0.015 3,protein:0.183 4±0.058 9,0.096 8± 0.015 7,0.147 5±0.0418)and negative control group(mR-NA:0.9132±0.0098,1.0032±0.0156,0.9327± 0.011 2,protein:0.187 9±0.042 3,0.088 9±0.0513,0.134 8±0.050 1),the mRNA(3.243 2±0.013 6,2.976 6 ±0.0213,1.585 9±0.019 8)and protein expressions(0.452 7±0.052 4,0.218 9±0.047 5,0.301 4±0.059 8)of IRAK4,NF-κB and anti-apoptosis protein BCL2 in MYD88L265P overexpression group were significantly increased,which was statistically significant(all P<0.05).Conclusion After overexpression of MYD88 L265P,the apoptosis rate of DLBCL cells decreased and the cell proliferation rate increased.The mechanism may be related to the mutation of MYD88 L265P gene,activation and amplification of NF-κB pathway,and pro-motion of the overexpression of antiapoptotic protein BCL2.

diffuse large B cell lymphomaMYD88 L265Pgene mutationNF-κB pathway

胡飘飘、宣成睿、杜华、李时荣、翁立新、海玲、乌云嘎、徐晓艳

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内蒙古医科大学基础医学院病理学教研室,呼和浩特 010059

南方医科大学南方医院病理科,广州 510080

弥漫大B细胞淋巴瘤 MYD88 L265P 基因突变 NF-κB通路

2024

临床与实验病理学杂志
安徽医科大学,中华医学会安徽分会

临床与实验病理学杂志

CSTPCD北大核心
影响因子:0.776
ISSN:1001-7399
年,卷(期):2024.40(1)
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