Liu Y, Qiao Y, Hu C, Liu L, Zhou L, Liu B, Chen H, Jiang X
Xuzhou Medical College Graduate Academy, Xuzhou, 221006, China.
Department of Radiation Oncology, Lianyungang First People's Hospital, No.182, Tongguan Road, Lianyungang City, 222002, Jiangsu Province, China.
Clin Transl Oncol. 2016 Feb;18(2):212-9. doi: 10.1007/s12094-015-1358-z. Epub 2015 Oct 12.
To investigate the role of the vascular endothelial growth factor receptor 2 (VEGFR2) in the proliferation, migration, invasion, and radiation-induced apoptosis of the non-small cell lung cancer (NSCLC) cell line Calu-1.
VEGFR2 gene was silenced by RNA interference in Calu-1 cells, and the expression of VEGFR2 was measured by qRT-PCR and Western blot analysis. The cells were divided into control, VEGF-treated, VEGFR2 knockdown, and VEGFR2 knockdown and VEGF-treated groups. A CCK8 assay and Transwell assay were performed to assess cell proliferation, migration, and invasion, respectively, after VEGFR2 knockdown. Western blot assays were used to detect signaling proteins downstream of VEGFR2. Cells in the groups listed above were also subjected to radiation treatment, followed by apoptosis analysis.
(1) RNA interference of VEGFR2 in Calu-1 cells reduced VEGFR2 mRNA (P < 0.01) and protein levels (P < 0.01). (2) VEGFR2 knockdown inhibited proliferation (P < 0.05), migration (P < 0.05), and invasion (P < 0.05) in Calu-1 cells. (3) VEGFR2 knockdown blocked the phosphorylation of protein kinase B (Akt, also known as PKB), extracellular regulated kinase (ERK) 1/2, and p38 mitogen-activated protein kinase (p38 MAPK) to various extent (P < 0.05), but did not change their total protein expression. (4) Knockdown of VEGFR2 suppressed HIF-1α protein synthesis (P < 0.05), and exacerbated apoptosis induced by radiation (P < 0.05).
VEGFR2 gene knockdown significantly suppressed a number of cellular activities in Calu-1 cells and increased radiation-induced cell death.
探讨血管内皮生长因子受体2（VEGFR2）在非小细胞肺癌（NSCLC）细胞系Calu-1增殖、迁移、侵袭及辐射诱导凋亡中的作用。
通过RNA干扰沉默Calu-1细胞中的VEGFR2基因，采用qRT-PCR和蛋白质印迹分析检测VEGFR2的表达。将细胞分为对照组、VEGF处理组、VEGFR2敲低组以及VEGFR2敲低+VEGF处理组。VEGFR2敲低后，分别采用CCK8法和Transwell法评估细胞增殖、迁移及侵袭能力。利用蛋白质印迹分析检测VEGFR2下游的信号蛋白。对上述分组的细胞进行辐射处理，随后进行凋亡分析。
（1）Calu-1细胞中VEGFR2的RNA干扰降低了VEGFR2 mRNA水平（P<0.01）和蛋白水平（P<0.01）。（2）VEGFR2敲低抑制了Calu-1细胞的增殖（P<0.05）、迁移（P<0.05）和侵袭（P<0.05）。（3）VEGFR2敲低在不同程度上阻断了蛋白激酶B（Akt，也称为PKB）、细胞外调节激酶（ERK）1/2和p38丝裂原活化蛋白激酶（p38 MAPK）的磷酸化（P<0.05），但未改变它们的总蛋白表达。（4）VEGFR2敲低抑制了HIF-1α蛋白合成（P<0.05），并加剧了辐射诱导的凋亡（P<0.05）。
VEGFR2基因敲低显著抑制了Calu-1细胞的多种细胞活性，并增加了辐射诱导的细胞死亡。
