Effect of silencing GRAMD1A and inhibiting STAT5 signaling pathway on radiosensitivity of hepatocellular carcinoma cell line Huh7
10.3760/cma.j.issn.0254-5098.2018.07.003
- VernacularTitle: 沉默GRAMD1A及抑制STAT5信号通路对肝癌细胞Huh7放射敏感性的影响
- Author:
Wei WANG
1
;
Shanbao KE
;
Mingbo LIU
;
Baiyu LI
;
Zhaojie WANG
Author Information
1. Department of Radiation Oncology, Henan Provincial People′s Hospital, Zhengzhou 450003, China
- Publication Type:Journal Article
- Keywords:
GRAMD1A;
Radiotherapy;
Hepatocellular carcinoma;
STAT5 signaling pathway;
SH-4-54(STAT inhibitor)
- From:
Chinese Journal of Radiological Medicine and Protection
2018;38(7):494-498
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of silencing GRAMD1A and inhibiting STAT5 signaling pathway on the radiosensitivity of Huh7 cells in order to provide new ideas for the clinical combined therapy of hepatocellular carcinoma.
Methods: The Huh7 cells silencing GRAMD1A was constructed by infecting lentivirus and verified by qPCR and Western blot. QPCR and luciferase reporter assays were used to detect the effect of silencing GRAMD1A on the expression of STAT5 and its downstream genes. Colony formation and apoptosis were detected to evaluate the effects of silencing GRAMD1A and STAT5 inhibitor SH-4-54 on cell radiosensitivity.
Results:After 2 Gy exposure of the constructed Huh7 cells, the colony formation ability of the silencing GRAMD1A combined irradiation group was significantly lower than that of the negative control combined irradiation group, and the difference was statistically significant (t=8. 494, P<0.05). Silence apoptosis in the GRAMD1A combined irradiation group was significantly increased compared with the negative control combined irradiation group (t=3.560, P<0.05). After silencing GRAMD1A, the radiosensitivity of Huh7 cells was significantly increased, and the expression of STAT5 and its downstream genes was significantly reduced in cells.The survival rate of the SH-4-54 inhibitor combined irradiation group was significantly lower than that of the dimethyl sulfoxide combined irradiation group, and the difference was statistically significant (t=8.660, P<0.05). SH-4-54 inhibited STAT5 after passage, the radiosensitivity of Huh7 cells was significantly increased.
Conclusions: Silencing GRAMD1A could significantly enhance the radiosensitivity of Huh7 cells via STAT5 signaling pathway, indicating that GRAMD1A plays an important role in the development and progression of HCC. This finding may provide a new target for HCC therapy.
