Li Jingjing, Xu Jinhua, Wu Guofeng, Ren Yajun, Wang Xue, Zhang Qianyun
Department of Pulmonary and Critical Care Medicine (PCCM) Ward II, Cangzhou Central Hospital, Cangzhou, China.
Department of Anesthesiology Ward II, Cangzhou Central Hospital, Cangzhou, China.
Chemotherapy. 2022;67(4):223-233. doi: 10.1159/000525231. Epub 2022 Jun 1.
Research suggests that circRNAs play important roles in non-small cell lung cancer (NSCLC). The function of hsa_circ_0068252 in NSCLC, especially in cisplatin (DDP) resistance and the mechanisms, was explored in this study.
NSCLC patient samples and two NSCLC cell lines along with corresponding DDP-resistant cell lines were used. Expression levels of circ_0068252 were detected. SiRNA for circ_0068252 and inhibitor for miRNA were used in all functional analyses. A co-culture system of NSCLC cells with CD8+ T cells was used. The cellular location of circ_0068252 was detected and its target miRNA was predicted and verified. Finally, the mechanism responsible for circ_0068252 function on PD-L1 was analyzed using luciferase reporter assay in the two DDP-resistant cell lines, as well as in the co-culture system. The cytotoxicity of T cells was detected by lactate dehydrogenase assay.
Our findings revealed that a high level of circ_0068252 was correlated with poor prognosis of NSCLC and DDP resistance. Knockdown of circ_0068252 could promote the sensitivity of DDP-resistant NSCLC cells to DDP. Moreover, knockdown of circ_0068252 could regulate the immune microenvironment which was mediated via CD8+ T cells. Finally, circ_0068252 could up-regulate PD-L1 expression by adsorbing miR-1304-5p.
The circ_0068252/miR-1304-5p/PD-L1 signal axis participates in the regulation of DDP resistance and immune escape of NSCLC cells. Our results suggest that circ_0068252 may be a potential diagnostic marker and therapeutic target for DDP-resistant NSCLC.
研究表明，环状RNA在非小细胞肺癌（NSCLC）中发挥重要作用。本研究探讨了hsa_circ_0068252在NSCLC中的功能，尤其是在顺铂（DDP）耐药性及其机制方面的作用。
使用NSCLC患者样本以及两种NSCLC细胞系及其相应的DDP耐药细胞系。检测circ_0068252的表达水平。在所有功能分析中使用针对circ_0068252的小干扰RNA（SiRNA）和针对微小RNA（miRNA）的抑制剂。采用NSCLC细胞与CD8+ T细胞的共培养系统。检测circ_0068252的细胞定位，并对其靶标miRNA进行预测和验证。最后，在两种DDP耐药细胞系以及共培养系统中，使用荧光素酶报告基因检测法分析circ_0068252对程序性死亡受体配体1（PD-L1）发挥作用的机制。通过乳酸脱氢酶检测法检测T细胞的细胞毒性。
我们的研究结果显示，高水平的circ_0068252与NSCLC的不良预后和DDP耐药性相关。敲低circ_0068252可提高DDP耐药NSCLC细胞对DDP的敏感性。此外，敲低circ_0068252可调节由CD8+ T细胞介导的免疫微环境。最后，circ_0068252可通过吸附miR-1304-5p上调PD-L1的表达。
circ_0068252/miR-1304-5p/PD-L1信号轴参与NSCLC细胞DDP耐药性和免疫逃逸的调控。我们的结果表明，circ_0068252可能是DDP耐药NSCLC的潜在诊断标志物和治疗靶点。
