Intensive Care Unit, The First People's Hospital of Huzhou, Huzhou, Zhejiang 313000, P.R. China.
Intensive Care Unit, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, P.R. China.
Mol Med Rep. 2018 Oct;18(4):3769-3779. doi: 10.3892/mmr.2018.9361. Epub 2018 Aug 8.
Lung cancer is one of the most prevalent malignancies worldwide; it has been ranked the most lethal type of cancer. Non‑small cell lung carcinoma (NSCLC) comprises >80% of all types of lung cancer. Although certain achievements have been made in the treatment of NSCLC, including the targeted gene drug epidermal growth factor receptor‑tyrosine kinase inhibitor (EGFR‑TKI), the five‑year survival rate of patients has not significantly increased. A previous study demonstrated that B7‑H5, a novel co‑stimulatory molecule in the B7 molecule family, was negatively correlated with EGFR in pancreatic cancer. Thus, in the present study, we aimed to investigate whether EGFR participates in immune evasion, probably through regulation of B7‑H5 expression. NCI‑H1299 NSCLCL cells were divided into control, mock, small interfering‑EGFR and EGFR‑TKI groups. The cell viability and apoptosis rate were analysed by a Cell Counting Kit‑8 assay and flow cytometry. The transforming growth factor (TGF)‑β and interleukin (IL)‑10 content was measured using an ELISA. The expression levels of EGFR, B7‑H5, Survivin, apoptosis regulator Bax, apoptosis regulator Bcl‑2 (Bcl‑2), TGF‑β, vascular endothelial growth factor (VEGF), IL‑10 and cyclooxygenase (COX)‑2 were assessed via quantitative PCR and western blotting. The activation of the tyrosine‑protein kinase JAK2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signalling pathway was detected using western blotting. The results demonstrated a notable negative correlation between EGFR and B7‑H5 expression levels in cancer tissues and cell lines. Inhibition of EGFR expression via gene silencing and EGFR inhibition markedly decreased cell viability and increased the apoptosis of NCI‑H1299 cells, by upregulating survivin and Bcl‑2 expression. The protein expression levels of TGF‑β, VEGF, IL‑10 and COX‑2 were additionally decreased, with weak activation of the JAK2/STAT3 signalling pathway. EGFR may be involved in immune evasion, possibly through regulation of B7‑H5 expression in NSCLC.
肺癌是全球最常见的恶性肿瘤之一；它已被列为最致命的癌症类型。非小细胞肺癌（NSCLC）占所有肺癌类型的>80%。尽管在 NSCLC 的治疗方面取得了一定的成就，包括靶向基因药物表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI），但患者的五年生存率并没有显著提高。先前的研究表明，B7 家族中的新型共刺激分子 B7-H5 与胰腺癌中的 EGFR 呈负相关。因此，在本研究中，我们旨在研究 EGFR 是否通过调节 B7-H5 表达参与免疫逃逸。将 NCI-H1299 NSCLC 细胞分为对照组、mock 组、小干扰 EGFR 组和 EGFR-TKI 组。通过细胞计数试剂盒-8 检测和流式细胞术分析细胞活力和细胞凋亡率。使用 ELISA 法测定转化生长因子（TGF）-β和白细胞介素（IL）-10 含量。通过定量 PCR 和 Western blot 检测 EGFR、B7-H5、Survivin、凋亡调节因子 Bax、凋亡调节因子 Bcl-2（Bcl-2）、TGF-β、血管内皮生长因子（VEGF）、IL-10 和环氧化酶（COX）-2 的表达水平。Western blot 法检测酪氨酸蛋白激酶 JAK2（JAK2）/信号转导和转录激活因子 3（STAT3）信号通路的激活情况。结果表明，在癌症组织和细胞系中，EGFR 与 B7-H5 表达水平之间存在显著的负相关。通过基因沉默和 EGFR 抑制抑制 EGFR 表达，可显著降低 NCI-H1299 细胞的活力并增加其凋亡，同时上调 Survivin 和 Bcl-2 的表达。TGF-β、VEGF、IL-10 和 COX-2 的蛋白表达水平也降低，JAK2/STAT3 信号通路的激活减弱。EGFR 可能参与免疫逃逸，可能通过调节 NSCLC 中的 B7-H5 表达。
