背景:染色体蛋白4(SMC4)的结构维持对于染色体组装和分离至关重要,但其在贲门腺癌(CA)中的作用和机制尚不清楚。
方法:首先通过蛋白质谱分析在20对CA肿瘤组织和邻近正常组织中检测SMC4的表达水平。然后使用蛋白质印迹分析评估CA细胞中的SMC4表达水平。通过CCK-8和克隆形成试验评价细胞增殖。使用划痕和transwell测试来研究细胞迁移和侵袭,而通过流式细胞术,我们检查了细胞凋亡和细胞周期的进展。使用蛋白质印迹研究了上皮-间质转化(EMT)和Wnt/β-catenin途径的调节作用。通过肿瘤发生实验研究SMC4对裸鼠肿瘤发展的影响。
结果:本研究显示CA组织和细胞中SMC4的过表达。SMC4敲除能显著抑制细胞增殖,移民和入侵,刺激细胞凋亡,诱导细胞周期阻滞在CA细胞的G0/G1期,并抑制体内肿瘤生长。此外,下调SMC4导致Bcl-2、CyclinD1、CDK4、CDK6、β-catenin、磷酸化GSK-3β,N-钙黏着蛋白,还有Vimentin,随着蛋白质水平的增加,即,Bax,裂开的caspase3和E-cadherin。当SMC4过表达时,这些影响被逆转了。
结论:SMC4可以促进CA的生物学进展,表明SMC4可能是该疾病的潜在治疗靶标。
Structural maintenance of chromosome protein 4 (
SMC4) is crucial for chromosome assembly and separation, but its role and mechanism in cardia adenocarcinoma (CA) are unknown.
SMC4 expression levels were initially detected by protein profiling in 20 pairs of CA tumor tissues and adjacent normal tissues. The level of
SMC4 expression in CA cells was then evaluated using a western blot analysis. Cell proliferation was evaluated by CCK-8 and clone formation tests. Scratch and transwell tests were used to investigate cell migration as well as invasion, while through the flow cytometry, we examined the cell apoptosis and progression of the cell cycle. The regulatory effects of the epithelial-mesenchymal transition (EMT) and the Wnt/β- catenin pathway were investigated using western blot. A tumorigenesis experiment was used to investigate the influence of SMC4 on tumor development in nude mice.
This study showed overexpression of
SMC4 in CA tissues and cells. Knockdown of SMC4 can significantly inhibit the proliferation, migration and invasion, stimulate cell apoptosis, induce cell cycle arrest in the G0/G1 phase of CA cells, and inhibit tumor growth in vivo. In addition, down-regulation of SMC4 resulted in decreased expression of Bcl-2, Cyclin D1, CDK4, CDK6, β-catenin, phosphorylated GSK-3β, N-cadherin, and Vimentin, with an increased level of proteins, i.e., Bax, cleaved-caspase3, and E-cadherin. When
SMC4 was overexpressed, these effects were reversed.
SMC4 can facilitate the biological progression of CA, suggesting that
SMC4 could be a potential therapeutic target for the disease.