Yuan Jun, Yang Jie, Wang Ruicang, Hao Hongling, Li Jie
Blood Specialty, Hebei General Hospital, Shijiazhuang, China.
Immunopharmacol Immunotoxicol. 2022 Jun;44(3):429-436. doi: 10.1080/08923973.2022.2052896. Epub 2022 Mar 30.
Diffuse large B-cell lymphoma (DLBCL) is a common lymphatic tumor in clinic. LncRNAs were reported to play a regulatory role in many cancers, including DLBCL. This study focused on the roles of NEAT1 in DLBCL.
Real-time quantitative polymerase chain reaction (RT-qPCR) was carried out to detect mRNA expression. StarBase as well as TargetScan was used to predict targeting relationships, which was confirmed by the Dual Luciferase Reporter Assay and RNA pull-down assay. Cell Counting Kit 8 (CCK-8) were applied to measure cell viability. Flow cytometry assay was applied to detect cell apoptosis. Western blotting assay was conduct to determine protein expression. Lactate dehydrogenase (LDH) release assay were applied to evaluated cell cytotoxicity.
NEAT1 was overexpressed in DLBCL patients. Knockdown of NEAT1 reduced the viability while enhanced the apoptosis of tumor cells. However, overexpression of NEAT1 exhibited an opposite effect. miR-495-3p was a target of NEAT1 and was decreased in DLBCL cells. However, inhibiting miR-495-3p reversed the effect of NEAT1 knock-down on DLBCL cells and induced the malignant behaviors of DLBCL cells. Moreover, NEAT1 functioned as a sponge of miR-495-3p to upregulate PD-L1.
Our study demonstrated that a NEAT1/miR-495-3p/PD-L1 axis regulated the development of DLBCL. Therefore, NEAT1 may be a potential biomarker for DLBCL.
弥漫性大B细胞淋巴瘤(DLBCL)是临床上常见的淋巴瘤。据报道,长链非编码RNA(lncRNAs)在包括DLBCL在内的许多癌症中发挥调节作用。本研究聚焦于NEAT1在DLBCL中的作用。
采用实时定量聚合酶链反应(RT-qPCR)检测mRNA表达。利用StarBase以及TargetScan预测靶向关系,并通过双荧光素酶报告基因检测和RNA下拉实验进行验证。应用细胞计数试剂盒8(CCK-8)检测细胞活力。采用流式细胞术检测细胞凋亡。进行蛋白质免疫印迹实验以测定蛋白质表达。应用乳酸脱氢酶(LDH)释放实验评估细胞毒性。
NEAT1在DLBCL患者中高表达。敲低NEAT1可降低肿瘤细胞活力,同时增强其凋亡。然而,过表达NEAT1则表现出相反的效果。miR-495-3p是NEAT1的靶标,且在DLBCL细胞中表达降低。然而,抑制miR-495-3p可逆转NEAT1敲低对DLBCL细胞的影响,并诱导DLBCL细胞的恶性行为。此外,NEAT1作为miR-495-3p的海绵,上调程序性死亡受体配体1(PD-L1)。
我们的研究表明,NEAT1/miR-495-3p/PD-L1轴调节DLBCL的发展。因此,NEAT1可能是DLBCL的潜在生物标志物。