Background

We investigated the expression of TLR genes and the effects of CpG ODN in Estrogen Receptor positive, Progesterone Receptor positive breast cancer cell line (T47D) and a triple-negative breast cancer cell line (MDA-MB-468) followed by studying the immunostimulatory activity of CpG oligonucleotides in breast cancer cell lines T47D and MDA-MB-468.

Materials and Methods

We evaluated the expression pattern of TLR genes (TLR1 to TLR9) in T47D and MDA-MB-468 cells using Real-time qPCR analysis. The intracellular TLR9 protein expression was studied by flow cytometry. The effect of CpG ODN on cell viability was tested using MTT assay. The relative expression of pro-inflammatory (IL6 and TNFα) and anti- inflammatory/immunosuppressive cytokines genes (IL10 and TGF beta1) were examined by Real-time qPCR.

Results

We found that MDA-MB-468 cells expressed TLR2, TLR3, TLR6, TLR8, and TLR9 genes and T47D cells expressed TLR3, TRL5, TLR8, and TLR9 genes. Stimulation of TLR9 in vitro with CpG significantly reduced the cell viability of T47D and MDA-MB-468 cells. IL6 cytokine gene expression was significantly reduced in both CpG treated T47D cells and MDA-MB-468 cells. TNFα gene expression was significantly reduced after treatment with CpG in MDA-MB-468 cells but not in T47D cells. IL10 and TGFβ1 expression were downregulated in CpG treated T47D cells. Whereas, IL10 and TGFβ1 were elevated in CpG treated MDA-MB-468 cells.

Conclusion

Our in vitro finding gives preliminary evidence that triggering TLR9 using CpG ODN decreases the cell proliferation and alters the pro-inflammatory cytokines in favor of inhibition of hormone receptor positive breast cancer cells T47D and triple negative breast cancer cells MDA-MB-468.

中文翻译：

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CpG寡核苷酸在激素受体阳性细胞系T47D和三阴性乳腺癌细胞MDA-MB-468中的Toll样受体（TLR）基因表达和免疫刺激作用。

背景

我们调查了TLR基因的表达以及CpG ODN在雌激素受体阳性，孕激素受体阳性乳腺癌细胞系（T47D）和三阴性乳腺癌细胞系（MDA-MB-468）中的作用，然后研究了免疫刺激活性乳腺癌细胞系T47D和MDA-MB-468中CpG寡核苷酸的表达

材料和方法

我们使用实时定量PCR分析评估了T47D和MDA-MB-468细胞中TLR基因（TLR1至TLR9）的表达模式。通过流式细胞术研究了细胞内TLR9蛋白的表达。使用MTT测定法测试了CpG ODN对细胞活力的影响。通过实时qPCR检查促炎（IL6和TNFα）和抗炎/免疫抑制细胞因子基因（IL10和TGF beta1）的相对表达。

结果

我们发现MDA-MB-468细胞表达TLR2，TLR3，TLR6，TLR8和TLR9基因，而T47D细胞表达TLR3，TRL5，TLR8和TLR9基因。CpG体外刺激TLR9会显着降低T47D和MDA-MB-468细胞的细胞活力。在经CpG处理的T47D细胞和MDA-MB-468细胞中，IL6细胞因子基因表达均显着降低。CpG处理后，在MDA-MB-468细胞中TNFα基因表达显着降低，而在T47D细胞中则没有。在CpG处理的T47D细胞中IL10和TGFβ1表达下调。而在CpG处理的MDA-MB-468细胞中，IL10和TGFβ1升高。

结论

我们的体外发现提供了初步证据，表明使用CpG ODN触发TLR9会降低细胞增殖并改变促炎性细胞因子，从而有利于抑制激素受体阳性乳腺癌细胞T47D和三阴性乳腺癌细胞MDA-MB-468。