Purpose
To establish orthotopic heterotransplanted model of retinoblastoma(RB)into the vitreous cavity of nude mice for experimental studies of gene therapies in vivo.
Methods
To prepare the concentrated cell suspensions of human RB cell line HXO-Rb44 and inject them into the vitreous cavities of 10 nude mice(20 eyes).The experimental mice were randomly divided into 2 groups:the first group,5 mice (10 eyes) were injected with 5 mu;l of the cell suspensions,and the second group,5 mice (10 eyes)with 10 mu;l．The transplanted RB was observed by inspection and ophthalmoscopy,and graded semi-quantitatively in vivo.Light and electrone microscopes were used for histopathological examination,ABC immunohistochemical methods for NSE and GFAP,and flow cytometric investigations for DNA index(DI) and s-phase fraction (SPF).
Results
All of the models of RB were successfully established by injecting the cell suspensions of human RB cell line into the mice vitreous cavities.The immunohistochemical reaction of the transplanted tumor cells to NSE was positive,and negative to GFAP．The DNA index was found to be more than 1.1,and the determination of SPF indicated the presence of proliferative ability of the transplanted RB cells.
Conclusion
The above findings reveal that the transplanted tumor cells were similar to those of the parent cells,hence the orthotopic heterotransplantation of RB via nude mice vitreous cavity injection is a superior method in establishing the animal model for the further experimental studies of gene therapies in vivo.
(Chin J Ocul Fundus Dis,1998,14:144-148)
Retinoblastoma susceptibility gene(Rb gene)is a tumor suppressor,which often deleted,mutated or inactivated in many malignant tumors.We construct an antisense Rb gene expression vector DOLRBAS and transfected it into human embryonic lung fibroblast(HEL cells)by electroporation technique.With the transient expression of antisense Rb gene the Rb protein synthesis is reduced and the growth rate of HEL cells in increased,but no coloney of HEL cells formed in soft agar.This result indicates that Rb gene can block cell division of HEL cells,but inactivation of Rb gene only is not sufficient for malignancy transformation of HEl cells. （Chin J Ocul Fundus Dis,1993,9:198-201）
