Radiological Biology
Effect of silencing GRAMD1A and inhibiting STAT5 signaling pathway on radiosensitivity of hepatocellular carcinoma cell line Huh7
Wei Wang, Shanbao Ke, Mingbo Liu, Baiyu Li, Zhaojie Wang
Published 2018-07-25
Cite as Chin J Radiol Med Prot, 2018, 38(7): 494-498. DOI: 10.3760/cma.j.issn.0254-5098.2018.07.003
Abstract
ObjectiveTo 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.
MethodsThe 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.
ResultsAfter 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.
ConclusionsSilencing 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.
Key words:
GRAMD1A; Radiotherapy; Hepatocellular carcinoma; STAT5 signaling pathway; SH-4-54(STAT inhibitor)
Contributor Information
Wei Wang
Department of Radiation Oncology, Henan Provincial People′s Hospital, Zhengzhou 450003, China
Shanbao Ke
Mingbo Liu
Baiyu Li
Zhaojie Wang
