Mu Yindong, Dong Jianjiang, Cui Hong, Hu Jiangping, Liang Jun, Yan Lei
Department of Histology and Embryology, Mudanjiang Medical University, Mudanjiang 157011, China.
Department of Pharmacy, Hongqi Hospital Affiliated to Mudanjiang Medical University, Mudanjiang 157011, China.
Evid Based Complement Alternat Med. 2022 Apr 22;2022:9299442. doi: 10.1155/2022/9299442. eCollection 2022.
This research aimed to explore the effect of Licochalcone-A (LCA) combined with gene on the proliferation, migration, and invasion of glioma U251 cells through the Wnt/-catenin signaling pathway. The glioma U251 cell line was taken as the research object, and the Rab23 overexpression plasmid was constructed. According to the treatment method, U251 cells were rolled into blank control group (BC), Rab23 overexpression plasmid transfection group (Rab23), 25 mol·L LCA treatment group (LCA), and Rab23 overexpression plasmid transfection combined with 25 mol·L LCA treatment group (Rab23 + LCA). Subsequently, the ability of cell proliferation, migration, and invasion of each group was detected by methyl thiazolyl tetrazolium (MTT) assay, scratch healing test, and Transwell cell invasion test, respectively. Western blot was implemented to detect the expression differences of cell proliferation antigen Ki-67, apoptosis-related proteins Bcl-2 and Bax, and Wnt/-catenin pathway-related proteins -catenin, glycogen synthase kinase-3 (GSK3), Axin2, and c-myc. The results showed the successful construction of Rab23 overexpression and stable transfection U251 cell line. After grouping and treatments, the cell proliferation, migration, and invasion ability of the Rab23 group, LCA group, and Rab23 + LCA group was substantially reduced relative to BC group ( < 0.05). In addition, the cell proliferation, migration, and invasion ability of Rab23 + LCA group decreased relatively more significantly. The expression levels of Ki-67, Bcl-2, -catenin, and c-myc in the Rab23, LCA, and Rab23 + LCA groups were greatly lower versus those of BC group. Moreover, the protein expression levels of Bax, GSK3, and Axin2 were considerably increased ( < 0.05), while the expression of protein in Rab23 + LCA group increased notably. These findings indicate that LCA combined with gene can inhibit the proliferation, migration, and invasion of glioma U251 cells through the Wnt/-catenin signaling and can promote cell apoptosis.
本研究旨在探讨甘草查尔酮A（LCA）联合基因通过Wnt/β-连环蛋白信号通路对胶质瘤U251细胞增殖、迁移和侵袭的影响。以胶质瘤U251细胞系为研究对象，构建Rab23过表达质粒。根据处理方法，将U251细胞分为空白对照组（BC）、Rab23过表达质粒转染组（Rab23）、25μmol·L LCA处理组（LCA）和Rab23过表达质粒转染联合25μmol·L LCA处理组（Rab23+LCA）。随后，分别采用甲基噻唑基四氮唑（MTT）法、划痕愈合试验和Transwell细胞侵袭试验检测各组细胞的增殖、迁移和侵袭能力。采用蛋白质免疫印迹法检测细胞增殖抗原Ki-67、凋亡相关蛋白Bcl-2和Bax以及Wnt/β-连环蛋白通路相关蛋白β-连环蛋白、糖原合酶激酶-3（GSK3）、Axin2和c-myc的表达差异。结果显示成功构建了Rab23过表达并稳定转染的U251细胞系。分组处理后，Rab23组、LCA组和Rab23+LCA组细胞的增殖、迁移和侵袭能力相对于BC组均显著降低（P<0.05）。此外，Rab23+LCA组细胞的增殖、迁移和侵袭能力下降相对更显著。Rab23组、LCA组和Rab23+LCA组中Ki-67、Bcl-2、β-连环蛋白和c-myc的表达水平均显著低于BC组。此外，Bax、GSK3和Axin2的蛋白表达水平显著升高（P<0.05），而Rab23+LCA组蛋白表达升高更为明显。这些研究结果表明，LCA联合基因可通过Wnt/β-连环蛋白信号通路抑制胶质瘤U251细胞的增殖、迁移和侵袭，并可促进细胞凋亡。
