Evaluation of the Anti-Tumor Activity of PD-1/PD-L1 Inhibitors Combined with Cisplatin Based on a Jurkat and Lung Cancer Cell Co-Culture Model
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摘要:
目的 构建Jurkat细胞与肺癌细胞体外共培养模型，评价PD-1/PD-L1抑制剂联合顺铂的体外抗肿瘤活性。 方法 将活化的Jurkat细胞分别与HCC95、A549细胞以不同比例共培养。实验分为顺铂单药组与联合用药组（顺铂分别联合卡瑞利珠单抗、替雷利珠单抗或特瑞普利单抗），干预24 h后，通过CCK-8法、流式细胞术、Western blot及ELISA检测细胞活性、凋亡率及PD-1、PD-L1、IL-2、IFN-γ的表达水平。 结果 成功建立了Jurkat/A549（5∶1）及Jurkat/HCC95（3∶1）体外共培养模型。给药后，与顺铂单药组相比，联合用药组肺癌细胞活性及PD-L1表达显著降低（P < 0.05），凋亡率显著升高（P < 0.01）；卡瑞利珠单抗/替雷利珠单抗联合组Jurkat细胞活性及PD-1表达、IFN-γ分泌水平均显著升高（P < 0.05）。 结论 PD-1/PD-L1抑制剂联合顺铂在肺癌体外模型中具有显著的协同抗肿瘤作用。 -
关键词:
- Jurkat /
- A549 /
- HCC95 /
- PD-1/PD-L1抑制剂 /
- 非小细胞肺癌
Abstract:Objective To establish an in vitro co-culture model of Jurkat cells and lung cancer cells, and to evaluate the in vitro anti-tumor activity of PD-1/PD-L1 inhibitors combined with cisplatin. Methods Activated Jurkat cells were co-cultured with HCC95 and A549 cells at different ratios, respectively. The experiment was divided into a cisplatin monotherapy group and combination therapy groups （cisplatin combined with camrelizumab, tislelizumab, or toripalimab）. After 24 hours of intervention, cell viability, apoptosis rate, and the expression/secretion levels of PD-1, PD-L1, IL-2, and IFN-γ were detected using CCK-8 assay, flow cytometry, Western blot, and ELISA. Results The in vitro co-culture models of Jurkat/A549 （ratio 5∶1） and Jurkat/HCC95 （ratio 3∶1） were successfully established. After drug administration, compared with the cisplatin monotherapy group, the viability and PD-L1 expression of lung cancer cells in the combination therapy groups were significantly decreased （P < 0.05）, and the apoptosis rates were significantly increased （P < 0.01）. Furthermore, in the camrelizumab and tislelizumab combination groups, the viability of Jurkat cells, as well as the PD-1 expression and IFN-γ secretion levels, were significantly increased （P < 0.05）. Conclusion PD-1/PD-L1 inhibitors combined with cisplatin exhibit a significant synergistic anti-tumor effect in the in vitro lung cancer model. -
Key words:
- Jurkat /
- A549 /
- HCC95 /
- PD-1/PD-L1 inhibitors /
- Non-small cell lung
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图 1 Jurkat细胞活化后PD-1、IL-2、IFN-γ表达水平（$ \bar x \pm s $，n = 3）
A：PD-1的表达情况及统计学分析；B：IFN-γ、IL-2表达统计学分析。
Figure 1. Expression levels of PD-1，IL-2，and IFN-γ in Jurkat cells post-activation（$ \bar x \pm s $，n = 3）
图 2 细胞活性变化（$ \bar x \pm s $，n = 3）
A：A549和活化或未活化Jurkat细胞共培养，A549细胞活性变化；B：Jurkat细胞活性变化；C：HCC95和活化或未活化Jurkat细胞共培养，HCC95细胞活性变化；D：Jurkat细胞活性变化；**P < 0.01。*P < 0.05。
Figure 2. Changes in cell activity（$ \bar x \pm s $，n = 3）
图 3 细胞活性变化（$ \bar x \pm s $，n = 3）
A：HCC95和活化或未活化Jurkat细胞共培养，HCC95细胞活性变化；B：Jurkat细胞活性变化；**P < 0.01。
Figure 3. Changes in cell activity（$ \bar x \pm s $，n = 3）
图 4 癌细胞凋亡情况（$ \bar x \pm s $，n = 3）
A：A549和活化或未活化Jurkat细胞共培养，A549细胞凋亡情况；B：HCC95和活化或未活化Jurkat细胞共培养，HCC95细胞凋亡情况；**P < 0.01。*P < 0.05。
Figure 4. Apoptosis of cancer cells（$ \bar x \pm s $，n = 3）
图 5 PD-1、PD-L1表达水平（$ \bar x \pm s $，n = 3）
A：A549和活化或未活化Jurkat细胞共培养，A549细胞PD-L1表达水；B：Jurkat细胞PD-1表达水平；C：HCC95和活化或未活化Jurkat细胞共培养，HCC95细胞PD-L1表达水平；D:Jurkat细胞PD-1表达水平；**P < 0.01。*P < 0.05。
Figure 5. The expression levels of PD-1 ，PD-L1（$ \bar x \pm s $，n = 3）
图 6 IL-2、IFN-γ分泌水平（$ \bar x \pm s $，n = 3）
A：A549和活化或未活化Jurkat细胞共培养，IL-2、IFN-γ的分泌水平；B：HCC95和活化或未活化Jurkat细胞共培养，IL-2、IFN-γ的分泌水平；**P < 0.01。
Figure 6. Secretion levels of IL-2 and IFN-γ （$ \bar x \pm s $，n = 3）
图 7 细胞活性变化（$ \bar x \pm s $，n = 3）
A：A549和Jurkat细胞共培养药物处理后，A549细胞活性变化；B：Jurkat细胞活性变化；C：HCC95和Jurkat细胞共培养药物处理后，HCC95细胞活性变化；D：Jurkat细胞活性变化；**P < 0.01。*P < 0.05。
Figure 7. Changes in cell activity（$ \bar x \pm s $，n = 3）
图 8 癌细胞凋亡情况（$ \bar x \pm s $，n = 3）
A：A549和Jurkat细胞共培养给药处理后，A549细胞凋亡情况； B：HCC95和Jurkat细胞共培养给药处理后，HCC95细胞凋亡情况；**P < 0.01。
Figure 8. Apoptosis of cancer cells（$ \bar x \pm s $，n = 3）
图 9 PD-1、PD-L1表达水平（$ \bar x \pm s $，n = 3）
A：A549和Jurkat细胞共培养给药处理后，A549细胞PD-L1表达水平；B：Jurkat细胞PD-1表达水平；C：HCC95和Jurkat细胞共培养给药处理后，HCC95细胞PD-L1表达水平；D：Jurkat细胞PD-1表达水平。
Figure 9. The expression levels of PD-1 and PD-L1（$ \bar x \pm s $，n = 3）
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