About 90% of cancer-related deaths are due to metastasis of cancer cells, and angiogenesis is a critical step in this process. sFLT01 is a novel fusion protein and a dual-targeting agent that neutralizes both VEGF and PlGF proangiogenic activities. GRP78 dual effect in tumor growth and angiogenesis could be activated under VEGF stimulation. The current study was designed to investigate the inhibitory impact of sFLT01 protein on VEGF/GRP78 axis. To this point, sFLT01 construct was synthesized, recombinant plasmid was expressed in eukaryotic host cells, sFLT01-HisTag protein was extracted and analyzed. The functional activity of sFLT01 on VEGF-enhanced tube formation and angiogenesis of HUVEC cells were examined. Eventually, the inhibitory impact of sFLT01 on growth, invasiveness, and migration of human prostate cancer cell line, DU145, was assessed. Real-time PCR evaluated the level of GRP78 and its effect on the downstream factors; matrix metallopeptidase proteins 2&9 (MMP2&9) along with tissue inhibitor of metalloproteinase proteins1&2 (TIMP1&2) under sFLT01 stimulation. According to the data, sFLT01 protein showed modulatory impact on proliferation, invasion, and migration of DU145 cells along with the potential of HUVECs angiogenesis. Real-Time PCR analysis depicted a significant downregulation in GRP78, MMP2 and MMP9 transcripts’ levels, and a subsequent elevation of TIMP1 and TIMP2 expression under sFLT01 stimulation was detected. Overall, these data indicated that the inhibitory impact of sFLT01 on cancer cells growth and invasiveness could be mediated through the modulation of VEGF/GRP78/MMP2&9 axis and activation of TIMPs.

中文翻译：

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sFLT01通过抑制VEGF/GRP78/MMP2&9轴调节前列腺癌DU145细胞的侵袭和转移

大约 90% 的癌症相关死亡是由于癌细胞的转移造成的，而血管生成是这一过程中的关键步骤。sFLT01 是一种新型融合蛋白和双重靶向剂，可中和 VEGF 和 PlGF 促血管生成活性。GRP78 在肿瘤生长和血管生成中的双重作用可以在 VEGF 刺激下被激活。目前的研究旨在研究 sFLT01 蛋白对 VEGF/GRP78 轴的抑制作用。至此，sFLT01构建体被合成，重组质粒在真核宿主细胞中表达，sFLT01-HisTag蛋白被提取和分析。检查了 sFLT01 对 VEGF 增强的 HUVEC 细胞管形成和血管生成的功能活性。最后，评估了 sFLT01 对人前列腺癌细胞系 DU145 的生长、侵袭和迁移的抑制影响。Real-time PCR评估了GRP78的水平及其对下游因素的影响；在 sFLT01 刺激下，基质金属肽酶蛋白 2&9 (MMP2&9) 以及金属蛋白酶蛋白 1&2 (TIMP1&2) 的组织抑制剂。根据数据，sFLT01 蛋白对 DU145 细胞的增殖、侵袭和迁移以及 HUVEC 血管生成的潜力具有调节作用。实时 PCR 分析显示 GRP78、MMP2 和 MMP9 转录物的水平显着下调，随后检测到 sFLT01 刺激下 TIMP1 和 TIMP2 表达升高。总体而言，这些数据表明 sFLT01 对癌细胞生长和侵袭的抑制作用可以通过调节 VEGF/GRP78/MMP2&9 轴和激活 TIMPs 来介导。