THE EXPRESSION OF TLR4, IFN-γ, TGF-β AND TNF-αLL LINE OF HUMAN SMALL CELL LUNG CARCINOMA NCI-H69 AND IN CISPLATIN-RESISTANT SUBLINE NCI-H69/CPR

I. Stupak
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Abstract

Aim: to investigate the effect of teichoic acid Staphylococcus aureus for expression of pro-inflammatory cytokines and of TLR4 in a human small cell lung carcinoma cell line NCI-H69, and cisplatin resistant subline NCI-H69/CPR. Methods. Incubation of cells with teichoic acid (1 ng/m) conducted for 2 days. Expression level of TLR4, TGF-β, INF-γ, TNF-α was evaluated by the real time PCR on 7500 Real-Time PCR System, using specific primers and fluorochrome SYBR Green. The reverse transcription reaction was performed with High-Capacity cDNA Reverse Transcription Kit carried out under the conditions: 25 °C - 10 min, 37 °C ‒ 120 min and 85 °C ‒ 5 min. Results. In cell line culture NCI-H69 addition of teichoic acid increased expression of TLR4 by 1.3 times, and IFN-γ – by 1,1 times. Expression of TGF-β and TNF-α was decreased 2.5 and 4.9 times respectively. In cell line culture NCI-H69/CPR the addition of teichoic acid inhibited the expression of all studied parameters. Expression TLR4 decreased by 4.2 times, IFN-γ – by 1.4 times. Expression TGF-β and TNF-α was depressed 1.6 and 1.2 times. The presented data indicate that teichoic acid of bacterial origin provided the effect of modulating the inflammatory effect in lung cancer cell culture, sensitive and resistant to cisplatin. Conclusions. Teichoic acid as a ligand of TLR4 modulates the expression of pro-inflammatory and anti-inflammatory cytokines in small cell lung cancer cell culture and suppresses the expression of TLR4 and all investigated cytokines in the cisplatin-resistant cell line NCI-H69.
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tlr4、ifn -γ、tgf -β和tnf -αll在人小细胞肺癌nci-h69及顺铂耐药亚系nci-h69 / cpr中的表达
目的:研究磷壁酸金黄色葡萄球菌对人小细胞肺癌细胞株NCI-H69和顺铂耐药亚系NCI-H69/CPR中促炎细胞因子和TLR4表达的影响。方法。用磷壁酸(1ng/m)培养细胞2天。采用特异性引物和荧光染料SYBR-Green在7500实时PCR系统上进行实时PCR检测TLR4、TGF-β、INF-γ、TNF-α的表达水平。使用高容量cDNA逆转录试剂盒在25°C-10分钟、37°C-120分钟和85°C-5分钟的条件下进行逆转录反应。在细胞系培养中,NCI-H69添加磷壁酸使TLR4的表达增加1.3倍,IFN-γ的表达增加1.1倍。TGF-β和TNF-α的表达分别下降2.5倍和4.9倍。在细胞系培养物NCI-H69/CPR中,磷壁酸的加入抑制了所有研究参数的表达。TLR4表达下降4.2倍,IFN-γ表达下降1.4倍。TGF-β和TNF-α的表达分别降低1.6和1.2倍。结果表明,细菌来源的磷壁酸在癌症细胞培养中具有调节炎症效应的作用,对顺铂敏感且耐药。结论。在小细胞肺癌癌症细胞培养中,作为TLR4配体的替酸调节促炎和抗炎细胞因子的表达,并抑制顺铂耐药细胞系NCI-H69中TLR4和所有研究的细胞因子表达。
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