Effect of hypoxia-inducible factor-1α on stemness and epirubicin sensitivity of HepG2 hepatoma cells / 临床肝胆病杂志

ObjectiveTo investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the stemness and epirubicin sensitivity of hepatoma cells. MethodsHepatoma cells were selected for experiment. HepG2 hepatoma cells transfected with HIF-1α overexpression plasmid were selected as experimental group, and those transfected with pcDNA3.1 empty plasmid were selected as control group; HepG2 cells alone were selected as HepG2 group. Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α; Western blot was used to measure the protein expression of HIF-1α; flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells. The three groups of cells were treated with epirubicin at different concentrations (0, 6.25, 12.5, 25, and 50 μmol/L) for 24 hours; MTT assay was used to measure cell viability, and flow cytometry was used to measure apoptosis after treatment with epirubicin (50 μmol/L). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for further comparison between two groups. ResultsCompared with the HepG2 group and the control group, the experimental group had a significant increase in the mRNA expression of HIF-1α (both P＜0.001), and Western blot showed high expression of HIF-1α in the experimental group. The percentage of CD133 cells was 0.040%±0.003% in the HepG2 group, 0.030%±0.010% in the control group, and 20.110%±0.600% in the experimental group, and the experimental group had a significantly higher positive rate of CD133+ than the HepG2 group and the control group (both P＜0.001). At an epirubicin concentration of 25 and 50 μmol/L, the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group (both P＜005). After the treatment with 50 μmol/L epirubicin for 48 hours, the experimental group had a significantly lower cell apoptosis rate than the HepG2 group (67.9%±2.5% vs 93.6%±1.5%, P＜0.001) and the control group (67.9%±2.5% vs 93.0%±1.2%, P＜0001). ConclusionHepG2 cells are successfully transfected with HIF-1α overexpression plasmid, and HIF-1α can increase the percentage of liver cancer stem cells and improve their resistance to epirubicin.

Expression of lymphokine-1 and clinical significance of p53 protein mutation in hilar cholangiocarcinoma / 中国综合临床

Objective:To analyze the expression and clinical significance of lymphoid enhancer factor-1 (LEF-1) and P53 protein in hilar cholangiocarcinoma.Methods:A total of 50 cases with hilarcholangiocarcinoma in the Tangshan People′s Hospital from March 2010 to December 2017 were retrospectively analyzed.P53 protein mutation was detected by immunohistochemistry.At the same time, the expression of LEF1 protein in cholangiocarcinoma and adjacent normal tissues was detected.The expression of LEF1 and its correlation with clinicopathological parameters were analyzed.At the same time, the relationship between p53 protein mutation and LEF1 protein expression and the prognosis of patients were discussed.Results:The positive expression rate of LEF1 in tumor tissues were 62.00%(31/50), which was higher than in adjacent tissues 36.00% (18/50). The difference was statistically significant (χ 2=6.763, P=0.016). There were 28 patients with TP53 protein mutation and 22 patients with wild type.The positive rate of LEF1 in TP53 mutant group was 75.00% (21/28), which was significantly higher than that in TP53 wild type group(45.45%, 10/22), and the difference was statistically significant(χ 2=4.565, P=0.03). The LEF1 expression was associated with tumor differentiation, TNM stage, lymph node metastasis and TP53 protein mutation (all P<0.05). LEF-1 expression was positively correlated with TP53 protein mutation ( r=0.294, P=0.04). The 1-year cumulative survival rate of patients with TP53 protein mutation was significantly lower than that of patients with TP53 protein wild type (60% vs.81%, P=0.0416). The 1-year cumulative survival rate of patients with LEF1 positive expression was also significantly lower than that of patients with negative expression (53% vs.82%, P=0.0180). Conclusion:LEF-1 is highly expressed in hilar cholangiocarcinoma, and patients with high expression have poor prognosis.The positive expression of LEF-1 in hilar cholangiocarcinoma patients with TP53 protein mutation was increased, and the prognosis of patients with TP53 protein mutation was poor.