抗TSLPR单克隆抗体对MCF-7和A549细胞活力、促凋亡基因表达和促炎细胞因子产生的影响。

Alyaa Rakha, Roba M Talaat, Eman A El-Maadawy, Adel A Gurguis
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引用次数: 0

摘要

背景:胸腺基质淋巴细胞生成素(TSLP)及其受体(TSLPR)在多种癌症细胞中表达。然而,它们在癌症发展中的作用并没有得到很好的界定。目的:研究抗TSLPR抗体对MCF-7和A549癌症细胞生存能力、促凋亡基因表达和促炎细胞因子产生的影响。材料和方法:将MCF-7和A549细胞暴露于抗TSLPR单克隆抗体24、48和72小时。MTT法检测其对细胞活力的影响。通过定量逆转录聚合酶链式反应(qRT-PCR)评估TP53、BAX和CASP3基因的表达水平。结果:抗TSLPR抗体处理MCF-7细胞后48小时和72小时细胞增殖略有刺激,24小时细胞毒性开始后,IL-6和TNF-α的产生显著降低。浓度为2.5μg/ml的抗TSLPR处理的细胞在24小时时BAX表达显著增加。在抗TSLPR处理的A549细胞中,没有观察到细胞计数的减少,并且在培养的48小时和72小时中,细胞增殖的轻微剂量依赖性刺激是明显的。在A549细胞中,用2.5μg/ml的抗TSLPR处理后,TP53、BAX和CASP3的表达显著增加。结论:抗TSLPR对MCF-7和A549细胞的细胞活力、促凋亡基因表达和促炎细胞因子(IL-6和TNF-α)产生的影响各不相同。
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EFFECT OF ANTI-TSLPR MONOCLONAL ANTIBODY ON VIABILITY, PROAPOPTOTIC GENES EXPRESSION, AND PRODUCTION OF PRO-INFLAMMATORY CYTOKINES IN MCF-7 AND A549 CELLS.

Background: Thymic stromal lymphopoietin (TSLP) and its receptor (TSLPR) are expressed in various cancer cells. However, their role in cancer development is not well defined.

Aim: To investigate the effects of anti-TSLPR antibody on the viability, proapoptotic genes expression, and production of pro-inflammatory cytokines in MCF-7 and A549 cancer cells.

Materials and methods: MCF-7 and A549 cells were exposed to anti-TSLPR monoclonal antibody for 24, 48, and 72 h. The effect on cell viability was examined by MTT assay. The expression levels of TP53, BAX, and CASP3 genes were evaluated by the quantitative reverse transcription polymerase chain reaction (qRT-PCR). Levels of interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), and transforming growth factor (TGF-β1) were measured by the enzyme-linked immunosorbent assay (ELISA).

Results: The treatment of MCF-7 cells with anti- TSLPR antibody slightly stimulates cell proliferation after 48 h and 72 h following initial cytotoxicity in 24 h with a significant reduction in IL-6 and TNF-α production. A significant increase in the BAX expression in anti-TSLPR treated cells at a concentration of 2.5 μg/ml at 24-h point was evident. In anti-TSLPR-treated A549 cells, no decrease in cell count was observed, and slight dose-dependent stimulation of cell proliferation was evident in 48 h and 72 h of culture. A significant increase in TP53, BAX, and CASP3 expression upon treatment with 2.5 μg/ml of anti-TSLPR was evident in A549 cells.

Conclusion: The effects of anti-TSLPR on cell viability, proapoptotic gene expression, and production of pro-inflammatory cytokines (IL-6 and TNF-α) vary in MCF-7 and A549 cells.

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