[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 B16RHSV, 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 B16RHSV, was generated. The permissibility of B16RHSV cells to oHSV infection was examined with green fluorescence protein (GFP)-expressing oHSV (oHSVGFP). To investigate the tumorigenic ability of both cells in vivo, 2×10(5) cells in 100 µl were subcutaneously inoculated into the right flanks of C57/BL mice.


RESULTS
In vitro, the B16RHSV mouse melanoma cells were shown by fluorescence microscopy capable of being infected by oHSVGFP. In vivo, the B16RHSV cells, like their wild type counterpart, grew to form melanoma in syngeneic mice.


CONCLUSIONS
A herpes simplex virus-permissive mouse melanoma cell line was established. Its tumorigenicity remained unchanged.
Cancer Institute & Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.
Xiu-fen
XF
Zhuang
Ai-ping
AP
Zhou
Gui-lan
GL
Shi
Xiang-ping
XP
Han
Jie
J
Li
Yu
Y
Zhang
You-hui
YH
Zhang
Shu-ren
SR
Zhang
Bin-lei
BL
Liu
D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D005784:Gene Amplification; D005822:Genetic Vectors; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology; D006801:Humans; D008545:Melanoma / Q000473:pathology* / Q000821:virology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D010957:Plasmids; D053240:Receptors, Tumor Necrosis Factor, Member 14 / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D047368:Tumor Burden
D053240:Receptors, Tumor Necrosis Factor, Member 14