[Generation of a herpes simplex virus-permissive mouse melanoma cell line B16RHSV].

Objective To generate an oncolytic herpes simplex virus （oHSV） permissive mouse melanoma cell line B16R^HSV, preserving the tumorigenic ability in syngeneic mice. Methods The herpes simplex virus entry mediator （HVEM） gene was amplified by PCR from human melanoma cell line A375, and cloned into pGEM-T Easy vector for sequencing. The HVEM gene was then cloned into pcDNA3 vector to generate pcDNA3-HVEM for transfection of mouse melanoma cell line B16-F10 cells. After that, the putative transfected cells were selected in full growth medium containing G418. The HVEM-expressing cells were isolated by immunomagnetic bead separation. The mouse melanoma cell line expressing oHSV receptor- HVEM, designated as B16R^HSV, was generated. The permissibility of B16R^HSV cells to oHSV infection was examined with green fluorescence protein （GFP）-expressing oHSV （oHSV^GFP ）. To investigate the tumorigenic ability of both cells in vivo, 2 ×10^5 cells in 100μl were subcutaneously inoculated into the right flanks of C57/BL mice. Results In vitro, the B16R^HSV mouse melanoma ceils were shown by fluorescence microscopy capable of being infected by oHSV^GFP. In vivo, the B16R^HSV cells, like their wild type counterpart, grew to form melanoma in syngeneic mice. Conclusion A herpes simplex virus-permissive mouse melanoma cell line was established. Its tumorigenicity remained unchanged. Key words: Simplexvirus ;  Melanoma;  Herpesvirus-entry mediator;  Mice