热休克蛋白70对X射线照射下wtp53蛋白的影响  

Impact of antisense heat shock protein 70 on expression and stability of wild type p53 protein after X-ray irradiation

作　　者：赵玫[1] 范云霞[1] 黄常志[1] 徐枫[1] 杜菲[1] 林梁[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤研究所病因室,北京100021

出　　处：《中华放射肿瘤学杂志》2001年第1期51-54,共4页Chinese Journal of Radiation Oncology

基　　金：国家自然科学基金资助项目!(39770 82 4)

摘　　要：目的　探讨反义热休克蛋白 70 (hsc70 )在X射线照射下对野生型p5 3(wtp5 3)的影响。方法　利用脂质体方法将本室构建的反义hsc70真核表达重组质粒pXAhsc70转染wtp5 3的MCF 7乳腺癌细胞 ,PCR证实外源DNA存在。Westernblot检测hsc70蛋白下降程度。X射线照射转染细胞后 ,检测wtp5 3蛋白的表达情况及半衰期变化。结果　转染重组质粒的细胞中hsc70蛋白下降 5 3% ;不同剂量X射线照射下 ,反义hsc70的存在能够提高wtp5 3的表达 ;4Gy相同剂量照射后 ,转染反义hsc70重组质粒的细胞中wtp5 3的半衰期延长。结论　反义hsc70能够上调wtp5 3的表达、稳定X射线照射下的wtp5 3蛋白并延长其半衰期 ;推测hsc70可能参与了wtp5Objective To investigate the effect of antisense heat shock protein (hsc70) on expression and stability of wild type p53 protein (wtp53) after X ray irradiation. Methods An eukaryotic recombination plasmid expressing antisense hsc70 RNA was transfected into MCF 7 human breast cancer cell line (wild type p53 ) by lipofectamine. The existence of foreign DNA was confirmed by means of PCR while suppression of hsc70 protein was monitored by Western blot. The expression and half life change of wild type p53 protein was evaluated after the definitive irradiation was implemented.Results Stable transfected MCF 7 cell strain expressing antisense hsc70 RNA was obtained. The expression of wtp53 was increased as compared to that of the control cell line which recieved the same dose of X ray and the half life period of wild type p53 was longer than that of the control cell after 4?Gy. Conclusions HSC70 plays an important role in stabilizing the wild type p53 protein expression with its molecular chaperon function in tumor cells irradiated. It is deduced that hsc70 is able to raise the expression of wtp 53 and to stabilize the wtp53 protein as well as to prolong its half life. It is expected that hsc70 may participate in the stabilization and degradation of wtp53.

关 键 词：反义RNA 野生型p53蛋白 热休克蛋白70 X射线照射 肿瘤 放射治疗 

分 类 号：R730.5[医药卫生—肿瘤]