Study of the Role of p53 in Bladder Carcinoma Cells

• Main Authors: Yu-Ling Li, 李豫玲
• Other Authors: Ming-Derg Lai
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/17016712678135552798

碩士 === 國立成功大學 === 生物化學研究所 === 82 === Bladder carcinoma is known to arise through a series of genetic alterations,including amplification of proto-oncogene c-erbB-2 (neu),point mutations on ras and p53,and homozygous inactiva- tion of tumor suppressor genes Rb and p53.In this study,we first determined the genetic alterations of p53 and ras in six cell lines,then introduced mutant p53 into a cell line containing endogenous wild-type p53 and study the effect of mutant p53 on the development of bladder carcinoma.In this six cell lines,wild-type p53 was detected in the TSGH-8301,ScaBER, TCC-SUP and BFTC-905 cells by EMSA analysis.Two forms of p53 was detected in BFTC-909,and less amount of p53 was detected in J82 by Western analysis.PCR-SSCP and PCR-DNA-sequencing revealed that both alleles of p53 gene in J82 were mutated, point mutation at codon 271 in one allele and 211bp deletion in the other.The results also showed point mutation at codon 241 in one allele in BFTC-909.In contrast,direct DNA sequencing indicated that no mutations occurred at condon 12,13 and 61 of Ha- and K-ras genes in the six cell lines.To study the effect of mutant p53 on the development of bladder carcinoma,we trans- fected pCDM8-p53/neo plasmid carrying the mutant p53 cDNA or pCDM8/neo plasmid into TSGH-8301 cell line which express only wild-type p53.With G418 selection, several transfectants and control transfectants were obtained.Among these,four trans- fectants and two control transfectants were further characte- rized. All four transfectants showed anchorage-independent growth in soft agar,while the control transfectants and the parental cell TSGH-8301 could not grow in soft agar. Furthermore, all four transfectants could form solid tumor in SCID mice(100%), while the control transfectants and TSGH-8301 couldn't(0%). These results indicated that mutant p53 plays an important role in the development of tumorigenicity of bladder carcinoma.