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沉默 COL1A2、COL6A3 和 THBS2 通过 PI3k-Akt 信号通路抑制胃癌细胞增殖、迁移和侵袭,同时促进细胞凋亡。

Silencing of COL1A2, COL6A3, and THBS2 inhibits gastric cancer cell proliferation, migration, and invasion while promoting apoptosis through the PI3k-Akt signaling pathway.

机构信息

Department of Gastroenterology, The First Affiliated Hospital of China Medical University, Shenyang, P.R. China.

出版信息

J Cell Biochem. 2018 Jun;119(6):4420-4434. doi: 10.1002/jcb.26524. Epub 2018 Feb 28.

Abstract

This study explores the effect of COL1A2, COL6A3, and THBS2 gene silencing on proliferation, migration, invasion, and apoptosis of gastric cancer cells through the PI3K-Akt signaling pathway. The gastric cancer microarray expression data (GSE19826, GSE79973, and GSE65801) was analyzed. Gastric cancer tissues and corresponding adjacent normal tissues were extracted from patients. Positive expression rate of PI3K, Akt, and p-Akt was measured with immunohistochemistry. Two cell lines, BGC-823 and SGC-7901, were transfected and cells were grouped into blank, negative control, COL1A2-shRNA, COL6A3-shRNA, and THBS2-shRNA groups. Expressions of COL1A2, COL6A3, and THBS2 in gastric cancer cells transfected with corresponding silencing sequences were evaluated by RT-qPCR and Western blot. MTT assay, Transwell, and cell scratch tests were conducted to evaluate cell proliferation, invasion, and migration capacity, respectively. Flow cytometry was used to evaluate cell cycle distribution and apoptosis. The positive expression of PI3K, Akt, and p-Akt was higher in gastric cancer tissues compared with adjacent normal tissues, and the mRNA expression of COL1A2, COL6A3, and THBS2 was increased in gastric cancer tissues. Akt, p-Akt, and PI3K expression drastically decreased in cells transfected with COL1A2, COL6A3, and THBS2 silencing sequences. Cells transfected with COL1A2, COL6A3, and THBS2 silencing sequences exhibited promoted apoptosis but inhibited proliferation, migration, and invasion. This study demonstrates that COL1A2, COL6A3, and THBS2 gene silencing inhibits gastric cancer cell proliferation, migration, and invasion while promoting apoptosis through the PI3K-Akt signaling pathway.

摘要

本研究通过 PI3K-Akt 信号通路探讨 COL1A2、COL6A3 和 THBS2 基因沉默对胃癌细胞增殖、迁移、侵袭和凋亡的影响。分析了胃癌微阵列表达数据(GSE19826、GSE79973 和 GSE65801)。从患者中提取胃癌组织及相应的癌旁正常组织,免疫组化法检测 PI3K、Akt 和 p-Akt 的阳性表达率。转染 BGC-823 和 SGC-7901 两个细胞系,分为空白组、阴性对照组、COL1A2-shRNA 组、COL6A3-shRNA 组和 THBS2-shRNA 组。采用 RT-qPCR 和 Western blot 检测相应沉默序列转染后胃癌细胞中 COL1A2、COL6A3 和 THBS2 的表达。MTT 法、Transwell 法和细胞划痕实验分别评估细胞增殖、侵袭和迁移能力,流式细胞术评估细胞周期分布和凋亡。与癌旁正常组织相比,胃癌组织中 PI3K、Akt 和 p-Akt 的阳性表达率较高,COL1A2、COL6A3 和 THBS2 的 mRNA 表达水平升高。COL1A2、COL6A3 和 THBS2 沉默序列转染后,Akt、p-Akt 和 PI3K 表达明显下降。COL1A2、COL6A3 和 THBS2 沉默序列转染的细胞凋亡增加,但增殖、迁移和侵袭受到抑制。本研究表明,COL1A2、COL6A3 和 THBS2 基因沉默通过 PI3K-Akt 信号通路抑制胃癌细胞增殖、迁移和侵袭,促进凋亡。

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