[Preliminary study of effect of antisense hsc70 effect on the stability of mutant p53 protein].
OBJECTIVE
To investigate the effect of antisense hsc70 RNA on the stability of mutant p53 protein.


METHODS
Using DNA recombination technique, retroviral recombinant expressing antisense hsc70 RNA was constructed and transfected into MCF7/Adr cells. After the existence of foreign DNA had been identified by the polymerase chain reaction (PCR) method, the protein level of hsc70 was detected by Western blot. The half-life of mutant p53 was detected by p53 stability assay.


RESULTS
With the stable expressive transfectant cell strains obtained through G418 selection, the foreign DNA in transfectant cells were confirmed by PCR method and the repression rate of hsc70 protein was 42%. The half-life of mutant p53 in MAc70 cell was 10 hours which was significantly lower than the control cells.


CONCLUSIONS
Antisense hsc70 RNA is able to lower the hsc70 protein level and significantly increase the instability of mutant p53 protein in MCF7/Adr human breast cancer cells.
Fan
Y
Y
Department of Etiology and Carcinogenesis, Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021, China.
Zhao
M
M
Huang
C
C
D015153:Blotting, Western; D002461:Cell Line, Transformed; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D016296:Mutagenesis; D016372:RNA, Antisense; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D018840:HSP70 Heat-Shock Proteins; D016372:RNA, Antisense; D016159:Tumor Suppressor Protein p53