[EB病毒LMP1通过mTORC1促进弥漫性大B细胞淋巴瘤细胞浆母细胞分化的机制]

[Mechanism about LMP1 of EB Virus Promoting Plasma Blast Differentiation of DLBCL Cell via mTORC1].

作者信息

Gao Jing-Jing, Zhu Xiong-Peng, Wang Ming-Qua, Lin Xing-Zhi, Zhuang Yan-Ling, Lin Hong-Jun

机构信息

Department of Blood Transfusion, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou 362000, Fujian Province， China.

Department of Haematology, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou 362000, Fujian Province， China. E-mail：

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2024 Feb;32(1):219-224. doi: 10.19746/j.cnki.issn.1009-2137.2024.01.035.

DOI:10.19746/j.cnki.issn.1009-2137.2024.01.035

PMID:38387925

Abstract

OBJECTIVE

To investigate possible mechanism on protien LMP1 expressed by EBV inducing plasmablast differentiation of DLBCL cell via the mTORC1 pathway.

METHODS

The expression levels of LMP1 protein, CD38 and the phosphorylation levels of p70S6K in EBV and EBV DLBCL cell lines were detected by Western blot. Cell lines overexpressing gene stablely were constructed and gene was silenced by RNAi. The expression of gene was verified by RT-qPCR. The expression levels of LMP1 and CD38 and the phosphorylation levels of p70S6K in each group were detected by Western blot.

RESULTS

Compared with EBVDLBCL cells, the expression of LMP1 was detected on EBV DLBCL cells ( =0.0008), EBV DLBCL cells had higher phosphorylation levels of p70S6K ( =0.0072) and expression levels of CD38( =0.0091). Compared with vector group, the cells of LMP1 group had higher expression levels of LMP1 and CD38 ( =0.0353; <0.0001), meanwhile molecular p70S6K was phosphorylated much more( =0.0065); expression of LMP1 mRNA was verified( <0.0001). Compared with si-NC group, expression level of LMP1 protein( =0.0129) was not detected and phosphorylated p70S6K disappeared of LMP1 group ( =0.0228); meanwhile, expression of CD38 decreased,although there was no significant difference ( =0.2377).

CONCLUSION

LMP1 promotes DLBCL cells plasmablast differentiation via activating mTORC1 signal pathway.

摘要

目的

探讨EBV表达的蛋白LMP1通过mTORC1途径诱导弥漫大B细胞淋巴瘤（DLBCL）细胞成浆细胞分化的可能机制。

方法

采用蛋白质免疫印迹法检测EBV及EBV DLBCL细胞系中LMP1蛋白、CD38的表达水平及p70S6K的磷酸化水平。构建稳定过表达基因的细胞系，并用RNAi使基因沉默。通过RT-qPCR验证基因的表达。采用蛋白质免疫印迹法检测各组中LMP1、CD38的表达水平及p70S6K的磷酸化水平。

结果

与EBV阴性DLBCL细胞相比，EBV阳性DLBCL细胞可检测到LMP1的表达（P = 0.0008），EBV阳性DLBCL细胞的p70S6K磷酸化水平更高（P = 0.0072），CD38表达水平更高（P = 0.0091）。与载体组相比，LMP1组细胞的LMP1和CD38表达水平更高（P = 0.0353；P < 0.000），同时分子p70S6K磷酸化程度更高（P = 0.0065）；验证了LMP1 mRNA的表达（P < 0.000）。与si-NC组相比，LMP1组未检测到LMP1蛋白表达（P = 0.0129），LMP1组磷酸化p70S6K消失（P = 0.0228）；同时，CD38表达下降，尽管无显著差异（P = 0.2377）。