Abstract
Objective: The study was designed to evaluate the role of Human cytomegalovirus (HCMV) infection on homebox (HOX) gene expression and the effects of overexpression of HOX genes on proliferation and apoptosis of vascular smooth muscle cells (VSMCs).Methods: Viral infection was verified by observation of cytopathic effects through inverted microscopy, viral particles by electron microscopy and HCMV IE gene amplification by RT-PCR. cDNA profiling technology was used to screen expression of HOX genes after HCMV infection in VSMCs. Abnormal expression of Haematopoietically-expressed homeobox (HHEX) was selected to construct over-expressed vector and transfected into VSMCs. The effects of over expression of HHEX on cell proliferation and apoptosis of VSMCs were assayed by flow cytometry. Apoptosis and proliferation-associated genes were also assayed by RT-PCR.Results: Multiple HOX gene expression levels had obvious changes after HCMV infection, among which expression of HHEX gene increased obviously at 24, 48, and 72 h after infection. Over expression of HHEX can promote VSMCs proliferation by promoting G0/G1 phase cells into S phase and inhibit VSMCs apoptosis. HHEX inhibited the expression of apoptosis-associated caspase 2 and caspase3 and promoted the expression of cell cycle-related genes such as CDK2 and CDK6, CyclinB2 and CyclinD2.Conclusion: HHEX over expression induced by HCMV infection closely associated with vascular proliferative diseases.
Highlights
Transplantation is the moving of an organ from one body to another or from a donor site to another location on the person’s own body, to replace the recipient’s damaged or absent organ
The human VSMCs were cultured in DMEM/F12 and inoculated with Human cytomegalovirus (HCMV)
RT-PCR for detecting HCMV IE gene and electron microscopy for detecting intact viral particles were used to study the infection of human VSMCs by HCMV
Summary
Transplantation is the moving of an organ from one body to another or from a donor site to another location on the person’s own body, to replace the recipient’s damaged or absent organ. With the continuous development of medical technology, transplantation has become an important and effective treatment after organ failure. The development of immunosuppressive agents and its application in clinical practice has largely been overcomed the acute rejection of transplantation. The long-term survival in patients has no obvious improvement (Tantravahi et al, 2007). This is mainly because chronic rejection deciding the long-term survival of patients has not been effectively resolved (Sarraj et al, 2014). Chronic rejection process is characterized by atrophy, fibrosis, and arteriosclerosis. Both immune and nonimmune mechanisms are likely
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