DUSP 促进 RPMI8226 骨髓瘤细胞衰老并抑制 TLR4 表达。

DUSP facilitates RPMI8226 myeloma cell aging and inhibited TLR4 expression.

机构信息

Department of Oncology, The Cancer Control Center of Nanchong Central Hospital, Nanchong, Sichuang, China.

出版信息

Eur Rev Med Pharmacol Sci. 2018 Sep;22(18):6030-6034. doi: 10.26355/eurrev_201809_15939.

DOI:10.26355/eurrev_201809_15939

PMID:30280787

Abstract

OBJECTIVE

Myeloma severely threatens public health, and molecular targeting treatment becomes the future perspective. Dual specificity phosphatases (DSUP) protein has multiple functions including modulating cell proliferation, differentiation, aging, and apoptosis. Whether DUSP can regulate myeloma cell is unclear. This study thus aimed to investigate the effect of DUSP on myeloma cell line RPMI8226 cell aging and provide evidence for the clinical treatment of myeloma.

MATERIALS AND METHODS

H2O2-induced aging model of myeloma cell line RPMI8226 was generated. DUSP over-expression plasmid or specific siRNA was transfected by liposome. Western blot was used to detect the expression of DUSP in RPMI8226 cells. Cell aging condition was evaluated by β-galactosidase assay. Aging proteins P53 and P16 expression levels, the activation of TLR4 signal pathway were tested by immunoblotting. TLR4 signal pathway was then suppressed by Verteporfin for testing RPMI8226 cell aging.

RESULTS

Growing levels of DUSP, aging proteins P53 and P16, with inhibition of TLR4 signal pathway were found in the H2O2-induced aging model of myeloma cell line RPMI8226. Transfection of DUSP over-expression plasmid or siRNA potentiated or inhibited the aging of RPMI8226 cells induced by H2O2 and suppressed or enhanced TLR4 signal pathway, respectively. Verteporfin, an inhibitor of TLR4, increased the level of P53 and aging of RPMI8226 cells.

CONCLUSIONS

DUSP facilitates H2O2-induced aging of myeloma cell line RPMI8226 and suppresses TLR4 expression, which provides academic basis for clinical intervention.

摘要

目的

骨髓瘤严重威胁公众健康，分子靶向治疗成为未来的研究方向。双特异性磷酸酶（DUSP）蛋白具有多种功能，包括调节细胞增殖、分化、衰老和凋亡。DUSP 是否能调节骨髓瘤细胞尚不清楚。本研究旨在探讨 DUSP 对骨髓瘤细胞系 RPMI8226 细胞衰老的影响，为骨髓瘤的临床治疗提供依据。

材料与方法

构建骨髓瘤细胞系 RPMI8226 的 H2O2 诱导衰老模型。通过脂质体转染 DUSP 过表达质粒或特异性 siRNA。Western blot 检测 RPMI8226 细胞中 DUSP 的表达。β-半乳糖苷酶染色检测细胞衰老状态。免疫印迹法检测衰老蛋白 P53 和 P16 的表达水平以及 TLR4 信号通路的激活情况。然后用 Verteporfin 抑制 TLR4 信号通路，检测 RPMI8226 细胞的衰老情况。

结果

在 H2O2 诱导的骨髓瘤细胞系 RPMI8226 衰老模型中，DUSP 表达水平升高，衰老蛋白 P53 和 P16 表达水平升高，TLR4 信号通路被抑制。DUSP 过表达质粒或 siRNA 的转染增强或抑制 H2O2 诱导的 RPMI8226 细胞衰老，并分别抑制或增强 TLR4 信号通路。TLR4 抑制剂 Verteporfin 增加了 P53 水平和 RPMI8226 细胞的衰老。