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Published on: November 11, 2014
[Preliminary study of effect of antisense hsc70 effect on the stability of mutant p53 protein]
1Department of Etiology and Carcinogenesis, Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100021, China.
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.
Conclusion:
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.
Insights
Antisense hsc70 RNA reduces heat shock cognate 70 kDa protein (hsc70) levels, destabilizing mutant p53 protein in breast cancer cells. This finding offers potential therapeutic strategies for p53-related cancers.
Area of Science:
- Molecular Biology
- Cancer Research
- RNA Therapeutics
Context:
- Mutant p53 protein accumulation is linked to cancer progression.
- Heat shock cognate 70 kDa protein (hsc70) may play a role in stabilizing mutant p53.
- Targeting protein stability is a strategy in cancer therapy.
Purpose:
- To determine the effect of antisense hsc70 RNA on mutant p53 protein stability.
- To investigate the potential of modulating hsc70 levels for cancer treatment.
Summary:
- Antisense hsc70 RNA was introduced into MCF7/Adr breast cancer cells using retroviral vectors.
- Polymerase chain reaction (PCR) and Western blot confirmed foreign DNA presence and hsc70 protein reduction (42%).
- The half-life of mutant p53 was significantly reduced to 10 hours in cells expressing antisense hsc70 RNA compared to controls.
Impact:
- Antisense hsc70 RNA effectively lowers hsc70 protein levels.
- This leads to significantly increased instability of mutant p53 protein.
- Suggests a therapeutic approach for human breast cancer by targeting mutant p53 instability.
