Abed Kahnamouei Shima, Baghaei Kaveh, Pakzad Parviz, Hashemi Mehrdad, Zali Mohammad Reza
Department of Biology, Faculty of Biological Sciences, North Tehran Branch, Islamic Azad University, Tehran, Iran.
Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Disease, Shahid Beheshti University of Medical Science, Tehran, Iran.
Gastroenterol Hepatol Bed Bench. 2020 Winter;13(Suppl1):S139-S144.
To acquire a deeper perception of EMT, we evaluated the expression of some candidate extra cellular matrix (ECM) proteins including THBS2, OSMR and CHI3L1 which were collected from RNA-seq bioinformatic analyses.
Gastric cancer (GC) is a major incident gastrointestinal cancer with a high rate of mortality. Metastasis is a challenging issue in gastric cancer treatment. Epithelial mesenchymal transition (EMT) of cancer cells is a complicated process controlled by different cells and molecular pathways regarded as an important step at the onset of metastasis.
AGS gastric cancer cell line was cultured and treated by TGF-β. EMT induction was verified by measuring the expression of E-cadherin, Snail, β-catenin and Vimentin genes by real time PCR. Then, following our previous study, we evaluated the expression of THBS2, OSMR and CHI3L1 genes in EMT induced cells by real time PCR.
Downregulation of E-cadherin and upregulation of Snail, β-catenin and Vimentin genes were verified in AGS treated cells in comparison with none-treated cells (P-value = 0.0355, P-value = 0.007, P-value = 0.0059, P-value = 0.0206 respectively). Also, upregulation of THBS2, OSMR and CHI3L1 were validated in these cells after EMT induction (P-value = 0.0147, P-value = 0.05, P-value = 0.05 respectively).
Our morphological and molecular results validated EMT induction by TGF- β cytokine in AGS gastric cancer cell line. Furthermore, significant upregulation of candidate genes including THBS2, OSMR and CHI3L1 verified the role of these proteins in gastric cancer invasiveness. However, further studies are needed for the validation of prognostic value of these markers.
为了更深入地了解上皮-间质转化(EMT),我们评估了一些候选细胞外基质(ECM)蛋白的表达,这些蛋白包括从RNA测序生物信息学分析中筛选出的THBS2、OSMR和CHI3L1。
胃癌(GC)是一种主要的胃肠道恶性肿瘤,死亡率很高。转移是胃癌治疗中的一个具有挑战性的问题。癌细胞的上皮-间质转化(EMT)是一个复杂的过程,受不同细胞和分子途径控制,被认为是转移起始的重要步骤。
培养AGS胃癌细胞系并用转化生长因子-β(TGF-β)处理。通过实时PCR检测E-钙黏蛋白、Snail、β-连环蛋白和波形蛋白基因的表达来验证EMT的诱导。然后,按照我们之前的研究,通过实时PCR评估EMT诱导细胞中THBS2、OSMR和CHI3L1基因的表达。
与未处理的细胞相比,在经TGF-β处理的AGS细胞中,E-钙黏蛋白表达下调,Snail、β-连环蛋白和波形蛋白基因表达上调得到验证(P值分别为0.0355、0.007、0.0059、0.0206)。此外,在这些细胞诱导EMT后,THBS2、OSMR和CHI3L1表达上调也得到验证(P值分别为0.0147、0.05、0.05)。
我们的形态学和分子学结果验证了TGF-β细胞因子在AGS胃癌细胞系中诱导EMT。此外,包括THBS2、OSMR和CHI3L1在内的候选基因显著上调,证实了这些蛋白在胃癌侵袭中的作用。然而,需要进一步研究来验证这些标志物的预后价值。