The effect of radionucleide hNIS gene therapy and lentivirus-mediated RNAi HexokinaseII in rat aortic vascular smooth muscle A7r5 cell line

Objectives Abnormal proliferation and migration of vascular smooth muscle cells(VSMCs) are important in pathogenesis of atherosclerosis. Sodium/iodide symporter(NIS) has been used radionucleide therapeutic gene for cancer. RNAi of HKII can be tool inhibit cell proliferation. Radionucleide hNIS gene therapy and HKII-shRNA for proliferation of VSMCs. This study was undertaken to clarify the effect of lentivirus-mediated HKII-shRNA gene expression on A7r5-NL cells.

Methods A7r5 cells were stably transfected with dual expression vector of hNIS and luciferase gene(pIRES-hNIS-Luc). Cells stably expressing the hNIS gene and luciferase gene were A7r5-NL produced, and the expression of the hNIS gene and luciferase gene was examined by I-125 uptake,luciferase assay and RT-PCR. Recombinant lentivirus producing HKII-shRNA was prepared. The effects of the lentivirus HKII-shRNA on protein and mRNA expression of HKII examined by western blot,RT-PCR. Cell growth was determined using MTT assay and effect of I-131 on A7r5-NL cells was also evaluation.

Results We established A7r5 cells which stably expressed hNIS and luciferase genes. Lentivirus-mediated RNAi effectively reduced endogenous HKII expression and downregulation of HKII in A7r5-NL cells and significantly inhibitied cell growth of A7r5-NL cells. A7r5-NL cells higher I-125 uptake and RLU compared to parent cell.

Conclusions Radionucleide hNIS gene therapy and the lentivirus-HKII-shRNA suppressed cell proliferation of A7r5-NL. Therefore both therapies can be applied to treat atherosclerosis or prevent recurrence of atherosclerotic process by inhibiting VSMC proliferation