Chen Borong, Zhu Zhipeng, Li Lulu, Ye Weipeng, Zeng Junjie, Gao Jin, Wang Shengjie, Zhang Liang, Huang Zhengjie
Department of Gastrointestinal Surgery, Xiamen Cancer Hospital, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian 361003, People's Republic of China.
Department of Clinical Medicine, Fujian Medical University, Fuzhou, Fujian 350004, People's Republic of China.
Onco Targets Ther. 2019 Jun 18;12:4667-4682. doi: 10.2147/OTT.S209734. eCollection 2019.
Using the gastric cancer cell line SGC7901, we constructed a cell line that overexpressed octamer-binding protein 4 (Oct4) and SRY-box 2 (Sox2) to explore the stem cell oncological and biological characteristics of these cells and to elucidate the mechanisms of Oct4 and Sox2 in cancer. A lentiviral vector containing the Sox2 gene was constructed and transfected into a gastric cancer cell line overexpressing Oct4 (SGC7901-Oct4) to obtain a stably transfected cell line (SGC7901-Oct4-Sox2). Oct4 and Sox2 expression was detected by RT-PCR and Western blotting. The proliferation, drug resistance, migration, and invasion abilities of the cells were assessed using in vitro (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium), drug resistance, scratch-wound migration, transwell migration, transwell invasion, and spherical clone formation assays, and their tumorigenic ability was assessed using a tumor formation experiment in mice. Compared with the control group, the expression of Oct4, Sox2, CD44, and E-cadherin was significantly higher in the group that overexpressed Oct4 and Sox2, while the expression of c-Myc and Klf4 did not significantly change. The proliferation, drug resistance, migration, and invasion abilities were significantly enhanced in the overexpression group, and the tumorigenic ability in mice was also significantly enhanced, with significantly increased tumor size and weight. The proliferation, drug resistance, migration, invasion, and tumorigenic abilities of SGC7901 cells overexpressing Oct4 and Sox2 were significantly improved. Oct4 and Sox2 play important roles in the proliferation, migration, invasion, and tumorigenicity of gastric cancer cells, and the two genes may be synergistic to a certain degree.
利用胃癌细胞系SGC7901,我们构建了一个过表达八聚体结合蛋白4(Oct4)和SRY盒2(Sox2)的细胞系,以探索这些细胞的干细胞肿瘤学和生物学特性,并阐明Oct4和Sox2在癌症中的作用机制。构建了一个包含Sox2基因的慢病毒载体,并将其转染到过表达Oct4的胃癌细胞系(SGC7901-Oct4)中,以获得稳定转染的细胞系(SGC7901-Oct4-Sox2)。通过RT-PCR和蛋白质免疫印迹法检测Oct4和Sox2的表达。使用体外(3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑)、耐药性、划痕伤口迁移、transwell迁移、transwell侵袭和球形克隆形成试验评估细胞的增殖、耐药性、迁移和侵袭能力,并使用小鼠肿瘤形成实验评估其致瘤能力。与对照组相比,过表达Oct4和Sox2的组中Oct4、Sox2、CD44和E-钙黏蛋白的表达显著更高,而c-Myc和Klf4的表达没有显著变化。过表达组的增殖、耐药性、迁移和侵袭能力显著增强,小鼠的致瘤能力也显著增强,肿瘤大小和重量显著增加。过表达Oct4和Sox2的SGC7901细胞的增殖、耐药性、迁移、侵袭和致瘤能力显著提高。Oct4和Sox2在胃癌细胞的增殖、迁移、侵袭和致瘤性中起重要作用,这两个基因可能在一定程度上具有协同作用。