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Abstract：
BACKGROUND: PD-L1 overexpression is commonly observed in various malignancies and is strongly correlated with poor prognoses for cancer patients. Moreover, PD-L1 has been shown to play a significant role in promoting angiogenesis and epithelial-mesenchymal transition (EMT) processes across different cancer types.
METHODS: The relationship between PD-L1 and vasculogenic mimicry as well as epithelial-mesenchymal transition (EMT) was explored by bioinformatics approach and immunohistochemistry. The functions of PD-L1 in regulating the expression of ZEB1 and the EMT process were assessed by Western blotting and q-PCR assays. The impact of PD-L1 on the migratory and proliferative capabilities of A549 and H1299 cells was evaluated through wound healing, cell invasion, and CCK8 assays following siRNA-mediated PD-L1 knockdown. Tube formation assay was utilized to evaluate the presence of VM structures.
RESULTS: In this study, increased PD-L1 expression was observed in A549 and H1299 cells compared to normal lung epithelial cells. Immunohistochemical analysis revealed a higher prevalence of VM structures in the PD-L1-positive group compared to the PD-L1-negative group. Additionally, high PD-L1 expression was also found to be significantly associated with advanced TNM stage and increased metastasis. Following PD-L1 knockdown, NSCLC cells exhibited a notable reduction in their ability to form tube-like structures. Moreover, the levels of key EMT and VM-related markers, including N-cadherin, MMP9, VE-cadherin, and VEGFA, were significantly decreased, while E-cadherin expression was upregulated. In addition, the migration and proliferation capacities of both cell lines were significantly inhibited after PD-L1 or ZEB1 knockdown.
CONCLUSIONS: Knockdown PD-L1 can inhibit ZEB1-mediated EMT, thereby hindering the formation of VM in NSCLC.
METHODS: The relationship between PD-L1 and vasculogenic mimicry as well as epithelial-mesenchymal transition (EMT) was explored by bioinformatics approach and immunohistochemistry. The functions of PD-L1 in regulating the expression of ZEB1 and the EMT process were assessed by Western blotting and q-PCR assays. The impact of PD-L1 on the migratory and proliferative capabilities of A549 and H1299 cells was evaluated through wound healing, cell invasion, and CCK8 assays following siRNA-mediated PD-L1 knockdown. Tube formation assay was utilized to evaluate the presence of VM structures.
RESULTS: In this study, increased PD-L1 expression was observed in A549 and H1299 cells compared to normal lung epithelial cells. Immunohistochemical analysis revealed a higher prevalence of VM structures in the PD-L1-positive group compared to the PD-L1-negative group. Additionally, high PD-L1 expression was also found to be significantly associated with advanced TNM stage and increased metastasis. Following PD-L1 knockdown, NSCLC cells exhibited a notable reduction in their ability to form tube-like structures. Moreover, the levels of key EMT and VM-related markers, including N-cadherin, MMP9, VE-cadherin, and VEGFA, were significantly decreased, while E-cadherin expression was upregulated. In addition, the migration and proliferation capacities of both cell lines were significantly inhibited after PD-L1 or ZEB1 knockdown.
CONCLUSIONS: Knockdown PD-L1 can inhibit ZEB1-mediated EMT, thereby hindering the formation of VM in NSCLC.
摘要：
背景：PD-L1过表达通常在各种恶性肿瘤中观察到，并且与癌症患者的不良预后密切相关。此外，PD-L1已被证明在促进不同癌症类型的血管生成和上皮间质转化（EMT）过程中起重要作用。
方法：通过生物信息学方法和免疫组织化学方法探讨PD-L1与血管生成拟态和上皮间质转化（EMT）的关系。通过蛋白质印迹和q-PCR测定评估PD-L1在调节ZEB1表达和EMT过程中的功能。通过伤口愈合评估PD-L1对A549和H1299细胞迁移和增殖能力的影响，细胞入侵，和siRNA介导的PD-L1敲低后的CCK8测定。利用管形成测定来评估VM结构的存在。
结果：在这项研究中，与正常肺上皮细胞相比,在A549和H1299细胞中观察到PD-L1表达增加。免疫组织化学分析显示，与PD-L1阴性组相比，PD-L1阳性组中VM结构的患病率更高。此外,研究还发现,PD-L1高表达与晚期TNM分期和转移增加显著相关.PD-L1敲除后,NSCLC细胞显示其形成管状结构的能力显著降低。此外，关键EMT和VM相关标记的水平，包括N-钙黏着蛋白,MMP9，VE-钙黏着蛋白，和VEGFA,显著下降，而E-cadherin表达上调。此外,PD-L1或ZEB1敲除后,两种细胞系的迁移和增殖能力均受到显著抑制.
结论：敲低PD-L1可以抑制ZEB1介导的EMT，从而阻碍了NSCLC中VM的形成。
方法：通过生物信息学方法和免疫组织化学方法探讨PD-L1与血管生成拟态和上皮间质转化（EMT）的关系。通过蛋白质印迹和q-PCR测定评估PD-L1在调节ZEB1表达和EMT过程中的功能。通过伤口愈合评估PD-L1对A549和H1299细胞迁移和增殖能力的影响，细胞入侵，和siRNA介导的PD-L1敲低后的CCK8测定。利用管形成测定来评估VM结构的存在。
结果：在这项研究中，与正常肺上皮细胞相比,在A549和H1299细胞中观察到PD-L1表达增加。免疫组织化学分析显示，与PD-L1阴性组相比，PD-L1阳性组中VM结构的患病率更高。此外,研究还发现,PD-L1高表达与晚期TNM分期和转移增加显著相关.PD-L1敲除后,NSCLC细胞显示其形成管状结构的能力显著降低。此外，关键EMT和VM相关标记的水平，包括N-钙黏着蛋白,MMP9，VE-钙黏着蛋白，和VEGFA,显著下降，而E-cadherin表达上调。此外,PD-L1或ZEB1敲除后,两种细胞系的迁移和增殖能力均受到显著抑制.
结论：敲低PD-L1可以抑制ZEB1介导的EMT，从而阻碍了NSCLC中VM的形成。
