Experimental Study
Effect and mechanism of JIB-04 on hepatoma cell line Huh7 cycle and apoptosis
Yang Chunyan, Zhong Chunqiang, Mo Cong, Bao Shiting
Published 2020-08-08
Cite as Chin J Exp Surg, 2020, 37(8): 1502-1504. DOI: 10.3760/cma.j.cn421213-20191120-01489
Abstract
ObjectiveTo study the effects of JIB-04 on the proliferation and apoptosis of hepatocellular carcinoma (HCC) Huh7 cells.
MethodsJIB-04, histone demethylase inhibitor, with different concentrations (0, 4, 8, 16 μmol/L) were used to treat Huh7 cells and normal hepatocyte L02 cells. Cell counting kit-8 (CCK-8) kit was used to detect cell proliferation of HCC cells. The flow cytometry was used to detect cell cycle and apoptosis of Huh7 cells. The expression of apoptotic proteins was detected by Western blotting in Huh7 cells.
ResultsJIB-04 had no obvious inhibitory effect on L02 cells. After treatment with 16 μmol/L JIB-04 for 48 h, the inhibition rate was only 5.6% and different concentrations of JIB-04 had different inhibitory effects on Huh7 cells. The inhibitory effects of JIB-04 on Huh7 cells were time- and concentration-dependent, and IC 50 was 3.5 μmol/L. The flow cytometry detection of hepatocellular carcinoma cells Huh7, cell cycle detection showed that the proportion of S phase and M phase of hepatocellular carcinoma cells were significantly reduced, while the proportion of G 1 phase was significantly increased. At the same time, with the increase of JIB-04 concentration, the apoptosis rate of Huh7 was significantly increased. Which were respectively (28.38±0.73)% after 48 h treatmenting with the highest JIB-04 concentration of 8 μmol/L. And there was statistically significant between control group and different concentration groups ( t=2.767, P<0.05); In the detection of hepatocarcinoma cell Huh7 apoptosis-related protein, the results suggest that the expression of protein B cell lymphoma/leukemia-2 associated X protein (bax) and p53 increased, and the expression of B cell lymphoma/leukemia-2 (bcl-2) was down-regulated.
ConclusionThe inhibitory effect of JIB-04 on normal hepatocytes was not obvious, but it had obvious inhibition on Huh7 hepatoma cells. JIB-04 has no obvious inhibitory effect on normal hepatocytes, which can inhibit the proliferation Huh7 hepatoma cells and interfere with their cycle distribution and promote apoptosis.
Key words:
Carcinoma, hepatocelular; JIB-04; Huh7; Apoptosis
Contributor Information
Yang Chunyan
Laboratory of Surgery, Clinical Skill Center, Guangdong Medical University, Zhanjiang 524001, China
Zhong Chunqiang
Department of Hepatobiliary Surgery, Huizhou Central People’s Hospital, Huizhou 516001, China
Mo Cong
Department of Hepatobiliary Surgery, Huizhou Central People’s Hospital, Huizhou 516001, China
Bao Shiting
Department of Hepatobiliary Surgery, the Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China
