Effects of neuropilin and tolloid-like 2 on proliferation, migration, cell cycle and apoptosis of gallbladder carcinoma cells and related mechanism
10.3760/cma.j.cn113884-20221216-00467
- VernacularTitle:神经纤毛以及类似透拉蛋白2对胆囊癌细胞增殖、迁移、细胞周期和凋亡的影响及相关机制研究
- Author:
Liwen DU
1
;
Junfeng YANG
;
Jian YE
;
Jiansheng LIU
Author Information
1. 山西医科大学第一医院肝胆胰外科，太原 030001
- Keywords:
Gallbladder neoplasms;
Neuropilin and tolloid-like 2;
Proliferation;
Migration;
Phosphatidylinositol-3 kinase;
Protein kinase B;
Epithelial-mesenchymal tran
- From:
Chinese Journal of Hepatobiliary Surgery
2023;29(5):357-363
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects and molecular mechanism of neuropilin and tolloid-like 2 (NETO2) on proliferation, migration, cell cycle, and apoptosis in gallbladder cancer (GBC).Methods:The NETO2 mRNA and protein expression in GBC-SD, ZJU-0430, NOZ GBC cells were detected by quantitative real-time polymerase chain reaction and Western blot. NETO2 overexpression and knockdown stable cell lines were constructed by plasmid transfection. Cell counting kit-8 assay, colony formation assay, transwell assay, flow cytometry and WB assay were performed to evaluate proliferation, migration, cell cycle, apoptosis, epithelial-mesenchymal transition (EMT) and changes of phosphatidylinositol-3 kinase/protein kinase B (PI3K/Akt) signaling pathway.Results:GBC-SD and ZJU-0430 cells with NETO2 gene overexpression and NOZ cells with NETO2 gene knockdown were effectively constructed. NETO2 overexpression in gallbladder cancer cell lines significantly improved cell proliferation and migration, advanced cell cycle progression from G0/G1 to S phase, and inhibited cell apoptosis. In the ZJU-0430 and GBC-SD cells, the clone number increased from (78.5±9.2), (217.0±6.4) to (213.5±10.3), (296.3±9.3)( t=10.98, 6.51; P=0.008, 0.023); The number of migrating cells increased from (198.6±8.4), (233.3±11.0) to (382.7±12.4), (379.0±7.3) ( t=16.98, 16.85, both P<0.001); The total apoptosis rate reduced from (29.7±0.9)%, (35.6±1.1)% to (19.2±0.5)%, (29.1±0.4)% ( t=9.74, 9.05; both P<0.001); The expression of EMT related proteins such as N-cadherin, Vimentin, Snail, and Slug were upregulated, while E-cadherin expression was downregulated. Phosphorylated PI3K (p-PI3K) and Akt (p-Akt) protein expression were significantly increased (all P<0.05). In contrast, NETO2 knockdown had the opposite effect on all these parameters. Conclusion:NETO2 influences the EMT process by regulating the PI3K/Akt signaling pathway, thus promotes GBC cell proliferation, migration and cell cycle progression, and inhibits cancer cell apoptosis.
