LTBP4介导的TGFβ_(1)信号通路在胃腺癌增殖中的作用

The role of LTBP 4-mediated TGFβ_(1) signaling in the proliferation of stomach adenocarcinoma

目的探讨潜在转化生长因子结合蛋白4(LTBP4)对胃腺癌肿瘤增殖的影响。方法检索公共数据集及本院胃腺癌患者,分析LTBP4在胃腺癌肿瘤组织及其癌旁组织中的表达。构建LTBP4过表达载体并转入MKN-45细胞中,检测细胞凋亡、增殖、细胞周期、细胞迁移、转化生长因子β_(1)(TGFβ_(1))信号通路与肿瘤体内增殖的改变。在公共数据集中,对在LTBP4低表达组中高表达的基因进行通路富集,并检测MKN-45细胞在经过瑞格菲尼处理后TGFβ_(1)信号通路的改变。同时应用瑞格菲尼处理MKN-45 LTBP4短发夹RNA(shRNA)细胞系,检测细胞体外、体内增殖的改变。结果在公共数据集与本院胃腺癌患者中,LTBP4均在胃腺癌肿瘤组织中低表达,且与患者总体生存率相关。LTBP4过表达后,细胞凋亡增加,细胞增殖、细胞迁移受限,细胞周期中G1期出现阻滞,TGFβ_(1)信号通路受到抑制,且肿瘤的体内增殖活性降低。血管内皮生长因子(VEGFR)、转化生长因子β_(1)(TGFβ_(1))信号通路均在LTBP4低表达的肿瘤组织中明显富集,且VEGFR抑制剂瑞格菲尼可逆转LTBP4表达,抑制TGFβ_(1)通路,并阻断MKN-45 LTBP4 sh RNA细胞的体内与体外增殖。结论LTBP4在胃腺癌中低表达,可通过激活TGFβ_(1)信号通路发挥对胃腺癌的调控作用。

Objective To clarify the effect and corresponding mechanism of Latent transforming growth factor binding protein 4(LTBP4)on the proliferation of stomach adenocarcinoma(STAD).Methods In the public dataset and in-house STAD patients,detecting the expression of LTBP4 in STAD tumor tissue and paracancerous tissue.The LTBP4 overexpression vector was constructed and transferred into MKN-45 cells to detect the alteration of cell apoptosis,proliferation,cell cycle,cell migration,transforming growth factorβ_(1)(TGFβ_(1))signaling pathway and tumor proliferation in vivo.In the public dataset,detecting the pathway enrichment in the up-regu Lated genes in LTBP4-low tissue.Detected the change of TGFβ_(1)signaling in MKN-45 after treat with Regorafenib.In the meantime,MKN-45 LTBP4 short hairpin RNA(shRNA)cells were treated with Regorafenib,and change of tumor proliferation in vitro,in vivo were quantified.Results In the public dataset and in-house patients,LTBP4 was low expressed in STAD tumor tissue,and correlated with overall survival rate.After LTBP4 was overexpressed,cell apoptosis was increased,cell proliferation,cell migration was limited,and G1 arrest in cell cycle was induced,TGFβ_(1)signaling was down-regulated,and tumor proliferation in vivo was decreased.Vascular endothelial growth factor(VEGFR)and TGFβ_(1)signaling was enriched in the LTBP4-low tissue,and VEGFR inhibitor Regorafenib could reverse LTBP4 expression and inhibit TGFβ_(1)signaling,and block the tumor proliferation in vitro and in vivo in MKN-45 LTBP4 shRNA cells.Conclusion LTBP4 is downregulated in stomach adenocarcinoma and exerts regulatory effects on stomach adenocarcinoma through activation of the TGFβ_(1)signaling pathway.