Basic Research Articles
Effects of neuropilin and tolloid-like 2 on proliferation, migration, cell cycle and apoptosis of gallbladder carcinoma cells and related mechanism
Du Liwen, Yang Junfeng, Ye Jian, Liu Jiansheng
Published 2023-05-28
Cite as Chin J Hepatobiliary Surg, 2023, 29(5): 357-363. DOI: 10.3760/cma.j.cn113884-20221216-00467
Abstract
ObjectiveTo investigate the effects and molecular mechanism of neuropilin and tolloid-like 2 (NETO2) on proliferation, migration, cell cycle, and apoptosis in gallbladder cancer (GBC).
MethodsThe 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.
ResultsGBC-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.
ConclusionNETO2 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.
Key words:
Gallbladder neoplasms; Neuropilin and tolloid-like 2; Proliferation; Migration; Phosphatidylinositol-3 kinase; Protein kinase B; Epithelial-mesenchymal transition
Contributor Information
Du Liwen
Department of Hepatobiliary and Pancreatic Surgery, the First Hospital of Shanxi Medical University, Taiyuan 030001, China
Yang Junfeng
Department of Hepatobiliary and Pancreatic Surgery, the First Hospital of Shanxi Medical University, Taiyuan 030001, China
Ye Jian
Department of Hepatobiliary and Pancreatic Surgery, the First Hospital of Shanxi Medical University, Taiyuan 030001, China
Liu Jiansheng
Department of Hepatobiliary and Pancreatic Surgery, the First Hospital of Shanxi Medical University, Taiyuan 030001, China
