Synergistic effect of interventing insulin-like growth factor-Ⅰ receptor activation combined with anti-cancer drugs in inhibiting the proliferation of hepatocellular carcinoma cells

• Main Authors: CAI Yin, YAO Min, WANG Li
• Format: Article
• Language: zho
• Published: Editorial Department of Journal of Clinical Hepatology 2016-08-01
• Series: Linchuang Gandanbing Zazhi
• Online Access: http://www.lcgdbzz.org/qk_content.asp?id=7607

ObjectiveTo investigate the intervention of gene transcription of insulin-like growth factor-Ⅰ receptor (IGF-ⅠR) and its synergistic effect with anti-cancer drugs in inhibiting the proliferation of hepatocellular carcinoma (HCC) cells. MethodsThe HBV-positive HCC PLC/PRF/5 and HBV-negative Bel-7404 cells were transfected with the efficient plasmid pGPU6/GFP/Neo-IGF-ⅠR-shRNA. Fluorescent quantitative RT-PCR and Western blot were used to measure mRNA and protein expression, the Cell Counting Kit-8 was used to analyze cell proliferation, and flow cytometry and Annexin-V-PE/7-ADD were used to analyze cell cycle and apoptosis. The t-test was used for comparison of continuous data between groups, the Fisher′s exact test was used for comparison of categorical data between groups. ResultsThe efficiency of IGF-ⅠR shRNA transfection was 71% in HCC PLC/PRF/5 cells and 90% in Bel-7404 cells, and both cells showed reductions in the mRNA and protein expression of IGF-ⅠR. The intervention group showed a significant inhibition compared with the negative control group, and the 72-hour inhibition rates of Bel-7404 cells and PLC/PRF/5 cells showed significant differences between the two groups (inhibition rates of Bel-7404 cells: 615%±17%vs 112%±09%, t=5.493, P＜0.05; inhibitionrates of PLC/PRF/5 cells: 639%±39%vs 95%±11%, t=19.244, P＜0.001). The intervention group showed a significantly higher apoptosis rate of Bel-7404 cells than the blank control group (35.96% vs 12.16%, P＜0.001) and the negative control group (3596% vs 943%, P＜0.001), as well as a significantly higher apoptosis rate of PLC/PRF/5 cells than the blank control group (4484% vs 662%, P＜0.001) and the negative control group (44.84% vs 4.02%, P＜0.001). The co-intervention group showed significantly higher percentages of Bel-7404 cells and PLC/PRF/5 cells in G0/G1 phase than the negative control group (59.0%±1.3% vs 484%±0.8%, t=12.032, P＜0.001; 65.4%±0.5% vs 53.5%±0.7%, t=22.789, P＜0.001). The co-intervention group showed significantly lower expression of cyclinD1 in Bel-7404 cells and PLC/PRF/5 cells than the negative control group (59.6%±4.7% vs 900%±3.4%, t=7.389, P＜0.05; 39.9%±0.5% vs 90.2%±14.6%, t=4.876, P＜0.05). The OD value of Bel-7404 cells and PLC/PRF/5 cells showed a significant difference between the intervention group and the negative control group when the sorafenib concentration was 0, 2.5, 5, 10, and 20 nmol/L and the oxaliplatin concentration was 0, 5, 10, 20, and 40 μmol/L (all P＜0.05). ConclusionThe downregulation of IGF-Ⅰ R gene transcription has the synergistic effect of inhibiting HCC cell proliferation and improving drug susceptibility.