| SUI Jianli,ZHOU Pingkun,HOU Jianyi,et al.Construction of eukaryotic vector expressing antisense RNA of human DNA repair enzyme APE and its effects on cellular radiation response[J].Chinese Journal of Radiological Medicine and Protection,1999,19(1):11-14 |
| Construction of eukaryotic vector expressing antisense RNA of human DNA repair enzyme APE and its effects on cellular radiation response |
| Received:September 25, 1998 Revised:December 05, 1998 |
| DOI： |
| KeyWords:Apurinicapyrimidinic endonuclease(APE) DNA repair Antisense nucleic acids Radiosensitization Hydrogen peroxide |
| FundProject:国家自然科学基金 |
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| Abstract:: |
| Objective To clone the cDNA encoding human apurinic/apyrimidinic endonuclease gene(hape), and to study the effects of its antisense nucleic acids on cytotoxicity of ionizing radiation and H 2O 2. Methods hape cDNA was cloned by RT-CPR. hape antisense RNA expression vector was constructed by inserting hape cDNA in reverse orientation into the mammalian cell expression vector pCI-neo. Gene transfection was mediated by lipofectamine. The cytotoxicity was assayed by means of cellular colony forming ability. Results 0.96kb of cDNA encoding hape was cloned from human embryonic lung (HEL) cells. Eukaryotic vector(pCIN/AShape) expression antisense hape RNA was constructed to transfect HOC8 cells. Stable HOC8 cell transfectants expressing hape antisense RNA were more sensitive to ionizing radiation and hydrogen peroxide than the control cells or the pCI-neo transfectants. Conclusion HAPE plays an important role in cellular protection against a wide range of cytotoxic DNA-damaging agents by its DNA repair or redox activities. hape locus is a potential target for the sensitization of radio or chemo therapy of tumors. |
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