Melanoma differentiation associated gene-7/interleukin-24 suppress the angiogenesis of hepatoma carcinoma cell line MHCC-97H by inhibiting endoplasmic reticulum stress proteins

Abstract

Objective To explore the mechanism of the melanoma differentiation associated gene-7/ interleukin-24 (MDA-7/IL-24) suppressing the angiogenesis of hepatoma carcinoma cell line MHCC-97H by inhibiting endoplasmic reticulum stress proteins.Methods By using the human hepatoma carcinoma cell line MHCC-97L as the experimental subject,MTT assay and flow cytometry were applied to examine the difference of tumor cell survival rate among different groups.The apoptosis of MHCC-97H cells was compared in the anoxic environments.The expression levels of MDA-7,extracellular signal-regulated kinase (ERK),E-cadherin (E-cad),vascular endothelial growth factor (VEGF),and matrix metalloproteinase (MMP)-9 mRNA were detected by using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).Western blotting was perform to observe the changes of MDA-7,protein kinase like endoplasmic reticulum kinase (PERK),E-cad,VEGF,and MMP-9 proteins among those groups.Results MTT showed that the growth inhibition rate of liver cells LO2 in hypoxia group and normoxia group was (12.60 ± 4.85)％ and (7.80 ± 0.61)％ respectively.Ad.MDA-7 had no the significant inhibitory effect on the growth of LO2 cells.Growth inhibition rate of MHCC-97H cells in hypoxia group and normoxia group was (5.10 ±0.09)％ and (4.80 ±0.05)％ respectively.Real-time PCR showed the expression of ERK,VEGF and MMP-9 mRNA in blank group and control blank group under hypoxia [(18.042 ± 5.915)/(17.512 ± 6.354),(22.654 ±5.976)/(22.587 ± 7.741),and (22.654 ± 2.789)/(22.587-± 8.551)] was higher than in normoxia group [(10.987 ±2.514)/(12.347 ±4.385),(15.321 ±4.262)/(15.634 ±8.521),and (15.321 ±6.384)/(15.634 ±4.423)].In hypoxic environments,E-cad mRNA expression in blank group and control blank group [(6.427 ± 0.512)/(6.042 ± 1.972)] was lower than in normoxia group [(10.987 ±2.514)/(12.347 ±4.385)].After Ad.MDA-7 interference,the expression of ERK,VEGF and MMP-9 mRNA was decreased [(10.987 ± 2.514)/(12.347 ± 4.385),(15.321 ± 4.262)/(15.634 ± 8.521),(15.321 ±6.384)/(15.634 ±4.423)],and that of E-cad was significantly increased [(19.573 ± 5.971)/(21.309 ± 8.544)] respectively in hypoxia group and normoxia group.Conclusion There was no significant difference in the growth inhibition rate of MHCC-97H cells between hypoxia and normoxia.MDA-7 significantly blocked PERK gene expression and inhibited HCC cell angiogenesis.Meanwhile,MDA-7 enhanced the expression of E-cad and inhibited the tumor cell metastasis synergically. Key words: Carcinoma, hepatocellular; Melanoma differentiation associated gene-7/interleukin24; Protein kinase like endoplasmic reticulum kinase; Angiogenesis; Metastasis