Department of Gastroenterology, The Fourth People's Hospital of Jinan, Jinan 250031, China.
Gastrointestinal Surgery, The Fourth People's Hospital of Jinan, Jinan 250031, China.
Biomed Res Int. 2021 Apr 15;2021:8814843. doi: 10.1155/2021/8814843. eCollection 2021.
This study is aimed at exploring the regulatory mechanism of 73HOXC-AS1 overexpression plasmid-activated Wnt-catenin classic signaling pathway and eukaryotic initiation factor 4A (eIF4AIII) expression increased by lentivirus-eIF4AIII-RNAi (44682-1) (LV-eIF4AIII-RNAi (44682-1)).
Focusing on the occurrence and progression of gastric cancer, the human gastric cancer cell line BGC823 (University Experimental Center) was taken as the research object and was transfected after subculture. According to the different ways of transfection, the cells were divided into the P1 group (LV-eIF4AIII-RNAi (44682-1) overexpressed plasmid), the P2 group (pcDNA-HOXC-AS1 overexpressed plasmid), the P3 group (LV-eIF4AIII-RNAi (44682-1) + pcDNA-HOXC-AS1), and the P4 group (no transfection, control group). Cell proliferation was detected by CCK-8 (Cell Counting Kit-8) assay, Western blotting was adopted to detect Wnt3a and P-GSK3 proteins, Transwell assay was adopted to detect the ability of cell migration and invasion, and cell cycle and apoptosis were detected by flow cytometry.
The results show that the protein expression levels of Wnt3a and P-GSK3 (glycogen synthase kinase-3) in the P1 and P4 groups were lower than those in the P2 and P3 groups ( < 0.05). The cell activity and clone number of BGC823 in the P3 group were higher than those in the P1, P2, and P4 groups ( < 0.05). The apoptosis rate of BGC823 cells in the P3 group was significantly higher than those in the P1, P2, and P4 groups ( < 0.05). The proportion of BGC823 cells in the P3 group at the S phase was significantly higher than those in the P1, P2, and P4 groups, while the proportion in the G2 phase was significantly lower than those in the P1, P2, and P4 groups ( < 0.05). The number of migrating and invading BGC823 cells in the P3 group was significantly higher than those in the P1, P2, and P4 groups, while the number of migrating BGC823 cells in the P4 group was significantly lower than those in the P1 and P2 groups ( < 0.05).
The 73HOXC-AS1 overexpression plasmid-activated Wnt-catenin classic signaling pathway and eIF4AIII expression increased by LV-eIF4AIII-RNAi (44682-1) could act together on BGC823 cells to improve cell proliferation activity, migration, and invasion; inhibit cell apoptosis; and prevent cells from entering the S phase.
本研究旨在探讨 73HOXC-AS1 过表达质粒激活 Wnt-连环蛋白经典信号通路和真核起始因子 4A（eIF4AIII）表达增加的调控机制，由慢病毒-eIF4AIII-RNAi（44682-1）（LV-eIF4AIII-RNAi（44682-1））引起。
以人胃癌 BGC823 细胞系（大学实验中心）为研究对象，聚焦于胃癌的发生和发展，在传代后进行转染。根据转染的不同方式，将细胞分为 P1 组（LV-eIF4AIII-RNAi（44682-1）过表达质粒）、P2 组（pcDNA-HOXC-AS1 过表达质粒）、P3 组（LV-eIF4AIII-RNAi（44682-1）+pcDNA-HOXC-AS1）和 P4 组（未转染，对照组）。通过 CCK-8（细胞计数试剂盒-8）检测细胞增殖，采用 Western blot 检测 Wnt3a 和 P-GSK3 蛋白，采用 Transwell 检测细胞迁移和侵袭能力，采用流式细胞术检测细胞周期和凋亡。
结果显示，P1 和 P4 组 Wnt3a 和 P-GSK3（糖原合成激酶-3）的蛋白表达水平均低于 P2 和 P3 组（<0.05）。BGC823 细胞在 P3 组的细胞活性和克隆数高于 P1、P2 和 P4 组（<0.05）。BGC823 细胞在 P3 组的凋亡率明显高于 P1、P2 和 P4 组（<0.05）。BGC823 细胞在 P3 组的 S 期比例明显高于 P1、P2 和 P4 组，而 G2 期比例明显低于 P1、P2 和 P4 组（<0.05）。BGC823 细胞在 P3 组的迁移和侵袭数量明显高于 P1、P2 和 P4 组，而 P4 组的迁移 BGC823 细胞数量明显低于 P1 和 P2 组（<0.05）。
LV-eIF4AIII-RNAi（44682-1）激活的 73HOXC-AS1 过表达质粒可共同作用于 BGC823 细胞，提高细胞增殖活性、迁移和侵袭能力；抑制细胞凋亡；阻止细胞进入 S 期。
