摘要
目的 探讨通过抑制蛋白激酶B（PKB、AKT）活性增加辐射敏感性，从而证实PKB活性是诱导SHG44细胞辐射抗性的机制之一。方法采用电穿孔法将PKB基因[pCMV5．HA-m／p-PKBa（PKB）、pCMV5-HA-PKBα-DD（T308D／S473D）（PKBD）]转染SHG44细胞；MTT法检测对照、PKB转染组及照射后各组细胞增殖率，激光共聚焦显微镜扫描技术观察对照、对照＋照射、PKB转染＋照射、PKBD转染＋照射组细胞凋亡及微细结构的变化；分析PKB转染的SHG44细胞增殖以及诱导凋亡的相关因素。结果含有外源性PKB的质粒成功转染了SHG44细胞并表达PKBmRNA，而对照组未见表达；转染后的SHG44细胞增殖率与对照组比较差异具有统计学意义（P〈0．05）；60Coγ），线照射能诱导SHG44细胞凋亡，以细胞核形态变化为特征；PKB＋照射组SHG44细胞形态完整，偶见凋亡细胞；而PKBD＋照射与对照＋照射组相比凋亡更显著。结论PKB转染SHG44细胞能抵抗辐射诱导的细胞凋亡，证实PKB活性与SHG44细胞辐射抗性存在一定的相关性。
Objective To explore the radiosensitivity of SHG44 cells increased by suppressing the protein kinase B （PKB）, in order to prove whether the PKB activity is related to the radioresistance of SHG44 cells. Methods PKB gene（pCMV5-HA-m/p-PKBα（PKB）,pCMV5.HA-PKBα-DD （T308D/S473D）（PKBD）） were transfected into SHG44 cells by electroporation, the cell proliferation rate was observed among the control, PKB transfected and irradiated groups by MTT assay. The laser confocal microscope was used to detect the changes of cell apoptosis and its microstructure in control, control ＋ radiation, PKB transfected ＋ radiation, PKBD transfected ＋ radiation group. The proliferation of PKB transfected SHG44 cells and the relative factors of inducing apoptosis were analyzed. Results The plasmid containing extrinsic PKB was successfully transfected into SHG44 cells and expressed PKB mRNA, while there was no expression in the control group; the proliferation rate of transfected SHG44 cells was significantly different from the control group （P 〈 0.05）. 60 Coγ rays could induce SHC,44 cell apoptosis with the changes of cell nuclei shape. The SHG44 cells transfected by PKB in the PKB ＋ control group were complete, with few apoptosis cells seen, while the apoptosis was more significant in PKBD ＋ irradiation group comparing to the control-irradiation group. Conclusions SHG44 cells transfected by PKB could resist the cell apoptosis induced by radiation, suggesting that there were some relations between PKB activity and SHG44 cells radioresistance.
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2007年第5期441-443,共3页
Chinese Journal of Radiological Medicine and Protection
基金
苏州大学医学发展基金重点项目资助（EE126031）
放射医学重点学科基金资助项目（ZY126001）
