Department of Gastroenterology, No. 1 Ward, ShiJiaZhuang No. 1 Hospital, No. 36, Fanxi Road, Chang'an District, Shijiazhuang, 050011, China.
Hum Cell. 2020 Jul;33(3):663-675. doi: 10.1007/s13577-020-00345-7. Epub 2020 May 14.
This study aims to investigate how Maspin affects the EMT and angiogenesis of gastric cancer (GC) cells via ITGB1/FAK pathway. Immunohistochemistry was used to evaluate the expressions of Maspin, ITGB1, FAK, E-cadherin, Vimentin, D2-40, and CD34 in GC and adjacent normal tissues from 160 patients. Then, the human GC cells with different degree of differentiation were transfected with Maspin CRISPR activation plasmid, ITGB1 siRNA and/or Maspin siRNA, followed by the following experiments, including qRT-PCR, western blotting, tube formation assay, Transwell assay and wound healing. GC tumor tissues manifested decreased Maspin with the activated ITGB1/FAK pathway. In tumor tissues, Maspin was negatively correlated with the expressions of ITGB1 and FAK, as well as Lauren's classification, differentiation degree, and TNM stage. Besides, Maspin was negatively related with lymphatic vessel density (LVD) and microvessel density (MVD), Vimentin and VEGF, but was positive correlated with E-cadherin. Maspin expression decreased, but ITGB1 and p-FAK expressions increased gradually in MKN-28 (well differentiated), SGC-7901 (moderate differentiated), and MKN-45 (poorly differentiated). Maspin CRISPR and ITGB1 siRNA increased E-cadherin with the decreased Vimentin, VEGF and bFGF, and the reductions of tube length. In comparison with the ITGB1 siRNA group, cells in the Maspin siRNA + ITGB1 siRNA group presented the more evident EMT and angiogenesis. Furthermore, ITGB1 siRNA reduced the malignancies of GC cells, which could be restored by Maspin siRNA. Maspin was downregulated in GC tissues, which could inhibit the EMT and angiogenesis by blocking the ITGB1/FAK pathway, thereby decreasing cell invasion and migration of GC.
本研究旨在探讨 Maspin 通过 ITGB1/FAK 通路对胃癌(GC)细胞 EMT 和血管生成的影响。采用免疫组织化学法检测 160 例患者 GC 组织及癌旁正常组织中 Maspin、ITGB1、FAK、E-cadherin、Vimentin、D2-40 和 CD34 的表达。然后,用 Maspin CRISPR 激活质粒、ITGB1 siRNA 和/或 Maspin siRNA 转染不同分化程度的人 GC 细胞,进行 qRT-PCR、western blot、管形成试验、Transwell 试验和划痕愈合试验。GC 肿瘤组织中 Maspin 表达降低,同时 ITGB1/FAK 通路被激活。在肿瘤组织中,Maspin 与 ITGB1 和 FAK 的表达以及 Lauren 分类、分化程度和 TNM 分期呈负相关。此外,Maspin 与淋巴管密度(LVD)和微血管密度(MVD)、Vimentin 和 VEGF 呈负相关,与 E-cadherin 呈正相关。MKN-28(高分化)、SGC-7901(中分化)和 MKN-45(低分化)中 Maspin 表达降低,而 ITGB1 和 p-FAK 表达逐渐升高。Maspin CRISPR 和 ITGB1 siRNA 增加了 E-cadherin,降低了 Vimentin、VEGF 和 bFGF,同时减少了管腔长度。与 ITGB1 siRNA 组相比,Maspin siRNA+ITGB1 siRNA 组 EMT 和血管生成更为明显。此外,ITGB1 siRNA 降低了 GC 细胞的恶性程度,而 Maspin siRNA 可恢复这种作用。GC 组织中 Maspin 表达下调,通过阻断 ITGB1/FAK 通路抑制 EMT 和血管生成,从而降低 GC 细胞的侵袭和迁移。