Assessment of Vascular endothelial growth factor and matrix mertaloproteinase-9 gene expression in human Hep-G2 cell line cultured in ORZ exposed PBMCs soup and co-cultured with ORZ exposed PBMCs

Abstract

Introduction: Bio-active peptides are digestion refractory amino-acids sequences that remain intact after digestion process and stimulate gut associated lymphoid tissue potentially. The aims of the current study is to evaluate the of the effects of ORZ on peripheral blood mono-nuclear cells (PBMCs) metabolic activity and also assessment of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP-9) gene expressions in HEP-G2 cell line after co-culture with ORZ stimulated PBMCs and ORZ treated PBMCs soup. Materials and Methods: In this study, PBMCs were isolated from 11 apparently healthy male volunteers after obtaining written informed consent. After isolation the effects of different concentrations of ORZ were evaluated on PBMCs proliferation. In addition after co-culture of HEP-G2 cell line with ORZ stimulated PBMCs and ORZ stimulated PBMCs soup, VEGF and MMP-9 gene expression were assessed in cancerous cells. Statisticl analysis were performed by SPSS ver. 16. P-value less than 0.05 considered as significan cut-point. Results: Evaluation of PBMCs proliferation that adjacent with bio active ORZ peptide two stimulated patterns were demonstrated among the study subjects. There were significant differences between the explored groups based on age (P< 0.039). Assessment of VEGF and MMP-9 gene expressions in HEP-G2 cell line after co- culture with ORZ treated PBMCs and ORZ stimulated PBMC soup showed that genes expression were affected only after 48 hours incubation of cancerous cells. In the group with lower mean age (SS group) after 48 hours incubation of HEP-G2 cell line with ORZ stimulated PBMCs the genes were overexpressed (P< 0.05). Similar results were seen after culture of the cancerous cells with ORZ stimulated PBMCs soup in SS group (P< 0.05). In the group with higher mean age (IS group) after 48 hours incubation of HEP-G2 cell line with ORZ stimulated PBMCs the genes expression were suppressed (P< 0.05). Similar results were seen after culture of the cancerous cells with ORZ stimulated PBMCs soup in IS group (P< 0.05). Conclusion: The results of the current study revealed that ORZ can change immune system response. According to production of peptide drugs based on ORZ structure for various usage such as induction of anorexia, long term administration of these peptide drugs may sensitize target population to cancers.