[Study on the dynamic changes of retinoblastoma gene of SO-Rb50 cell line].
OBJECTIVE
To study the dynamic changes of Retinoblastoma Gene of SO-Rb50 cell line.


METHODS
1) Southern blot hybridization was used to analyse the tumor cell DNA of 327th passage of SO-Rb50 cell line. 2) A promoter and 27 exons of Rb gene were screened exon-by-exon by using PCR-SSCP at 415th and 713th passages of SO-Rb50 cell line. 3) Three cell cloning strains named as MC2, MC3 and MC4 were isolated by single cell cloning technique from the SO-Rb50-775, and mutation of Rb gene were also screened exon-by-exon by using PCR-SSCP-HA in MC2-11, MC3-11, MC4-11 and MC3-138.


RESULTS
The 3.5 Kb, 2.9 Kb and 1.0 Kb bands were deleted in the DNA of SO-Rb50-327 tumor cells, showing the deletion of Rb gene in SO-Rb50 cell line. Exon23 of 451th passage cells decreased one band; but exon 25 of 713th passage cells decreased two bands, indicating that exon 25 had new mutation. PCR-SSCP-HA analysis of exon24 showed that MC4-11 and MC3-138 had abnormal bands, but MC2-11 and MC3-11 weren't found mutation of Rb gene. This result suggested that new mutation occurred to exon24 of MC3 during a long-term culture in vitro.


CONCLUSIONS
Retinoblastoma gene mutation of SO-Rb50 cell line had dynamic changes during a long-term culture.
Feng
G
G
Eye Research Institute, Zhongshan Ophthalmic Center, Sun Yat-sen University of Medical Sciences, Guangzhou 510060, China.
Li
H
H
Yi
Y
Y
Zheng
J
J
Zhang
Q
Q
Wang
X
X
Du
C
C
D005091:Exons; D016161:Genes, Retinoblastoma / Q000235:genetics*; D006801:Humans; D009154:Mutation; D018807:Polymorphism, Single-Stranded Conformational; D019572:Retinal Neoplasms / Q000235:genetics*; D012175:Retinoblastoma / Q000235:genetics*; D014407:Tumor Cells, Cultured