Experimental Study
Mechanism of N-acetyl amino galactosyl transferase-14 regulating the apoptosis of glioblastoma through extracellular signal-regulated kinase 1/2 signaling pathway
Shufeng Hao, Jianliang Wu, Chengye Wang, Liushuai Wang, Yuhua Hu
Published 2019-06-08
Cite as Chin J Exp Surg, 2019, 36(6): 1062-1065. DOI: 10.3760/cma.j.issn.1001-9030.2019.06.025
Abstract
ObjectiveTo study the effect of N-acetyl amino galactosyl transferase-14 (GalNAc-T14) on apoptosis of glioblastoma multiforme.
MethodsThe U87MG cell line of glioblastoma multiforme was cultured, and the constructed pcDNA3.1-T14 plasmid was transfected into U87MG cells. The cells were divided into untreated group (control), pcDNA3.1 (+ ) empty carrier-transfected group (NC1) and pcDNA3.1-T14-transfected group (T). Real-time quantitative polymerase chain reaction (qPCR) and Western blotting were used to detect the expression of GalNAc-T14, extracellular signal-regulated kinase 1/2 (ERK1/2), inhibitor of apoptosis protein-XL (bcl-XL) and the phosphorylation level of ERK1/2. Apoptosis was detected by flow cytometry with Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) double staining.
ResultsThe relative quality of GalNAc-T14, ERK1/2 and bcl-XL was 1 before transfection, and the relative expression of GalNAc-T14 was (15 393.43±30.35) in U87MG cells. Real-time quantitative PCR showed that the relative mRNA expression of ERK1, ERK2 and bcl-XL in NC1 group was (0.98±0.12), (0.91±0.05) and (0.89±0.11) respectively, and that in T group was (0.74±0.29), (0.68±0.31) and (0.68±0.23) respectively. The expression level of ERK1/2 and anti-apoptotic protein bcl-XL in T group was lower than that in control group. Western blotting analysis showed that the gray values of ERK1/2 in control, NC1 and T groups were (85.42±3.11), (86.25±4.25), and (66.36±4.25), and those of anti-apoptotic protein bcl-XL were (82.57±3.71), (79.52±3.78), and (46.85±4.52), respectively. The percentages of early apoptosis and late apoptosis of U87MG cells was (7.74±1.23)% and (10.08±2.41)%, respectively, when high expression of GalNAc-T14 was detected by Annexin V-FITC/PI double staining, which were higher than in the control group [(1.12±0.18)%, (0.51±0.14)%] and NC1 group [(1.37±0.12)%, (0.90±0.08)%, t=3.417, P<0.05].
ConclusionGalNAc-T14 promotes apoptosis of glioblastoma cells by inhibiting the expression of ERK1/2 and bcl-XL.
Key words:
Glioblastoma multiforme; N-acetyl amino galactosyl transferase-14; Extracellular signal-regulated kinase; Apoptosis
Contributor Information
Shufeng Hao
Department of Neurosurgery, the Chinese Medicine Hospital of Hebei Province, Shijiazhuang 050799, China
Jianliang Wu
Department of Neurosurgery, the Second Hospital of Heibei Medical University, Shijiazhuang 050799 China
Chengye Wang
Department of Neurosurgery, the Second Hospital of Heibei Medical University, Shijiazhuang 050799 China
Liushuai Wang
Department of Neurosurgery, the Second Hospital of Heibei Medical University, Shijiazhuang 050799 China
Yuhua Hu
Department of Neurosurgery, the Second Hospital of Heibei Medical University, Shijiazhuang 050799 China
