整合素A5调控FAK/Src/Rac1信号通路促进胃癌增殖、侵袭和黏附能力

Integrinα5 promotes the proliferation,invasion and adhesion of gastric cancer by regulating the FAK/Src/Rac1 signaling pathway

目的探讨整合素A5(ITGA5)在胃癌(GC)中的生物学作用和潜在机制。方法2019年1月至2020年12月,从浙江省人民医院接受手术切除的胃癌患者中收集35例胃癌组织[男21例,女14例,年龄(53.8±5.4)岁]以及35例匹配的癌旁组织,从北京百欧博伟生物技术公司购买胃癌和正常胃黏膜细胞。采用实时定量逆转录聚合酶链反应(qRT-PCR)、免疫组织化学、Western印迹法检测胃癌细胞中ITGA5、细胞黏附相关基因pFAK、pSrc、aRac1的mRNA和蛋白表达情况。细胞增殖检测(CCK-8)、Transwell侵袭实验、划痕愈合实验、细胞黏附实验检测胃癌细胞的表型。结果与正常胃黏膜细胞比较,ITGA5在胃癌细胞中高表达(1.00±0.26与1.23±0.27,P<0.05)。并且过表达ITGA5后能够通过FAK/Src/Rac1通路促进胃癌细胞增殖,吸光度(A)值由1.14±0.14升至1.61±0.14、迁移能力由(20.3±2.3)%提升至(56.4±6.1)%、侵袭能力由144.0±4.6提升至216.7±6.6,并增强与基质蛋白的黏附能力(99.0±8.5提升至152.0±12.3)(均P<0.05)。结论ITGA5在胃癌中高表达并作为促癌因子发挥作用,为寻找胃癌潜在治疗靶点提供了理论依据。

Objective To elucidate the biological role and potential mechanism of integrinα5(ITGA5)in gastric cancer(GC).Methods From January 2019 to December 2020,35 pairs of GC tissue[21 males and 14 females,aged(53.8±5.4)years]and matched adjacent tissue samples were collected from GC patients who underwent surgical resection in Zhejiang Provincial People′s Hospital.GC and normal gastric mucosa cells were purchased from Beijing Biobw Biotech Company.Quantitative real-time PCR(qRT-PCR),immunohistochemistry,Western blotting were performed to detect the mRNA and protein expression levels of ITGA5,cell adhesion-related genes(pFAK,pSrc,aRac1)in GC cells.Cell Counting Kit-8(CCK-8),Transwell invasion,wound healing and cell adhesion assays were conducted for GC cell phenotype detection.Results ITGA5 was highly expressed in GC compared with normal gastric mucosa cells(relative expression increased from 1.00±0.26 to 1.23±0.27,P<0.05).In addition,ITGA5 overexpression promoted the cell proliferation[from(1.14±0.14)OD to(1.61±0.14)OD],migration ability[from(20.3±2.3)%to(56.4±6.1)%],invasion ability(from 144.0±4.6 to 216.7±6.6),and adhesion ability of matrix protein(from 99.0±8.5 to 152.0±12.3)through FAK/Src/Rac1 signaling pathway in GC.(all P<0.05)Conclusions ITGA5 acts as a cancer-promoting factor in GC.The current study provides theoretical evidence for probing the novel molecular targets for the treatment of GC.