摘要
目的研究热休克预处理对高温致V79细胞DNA双链断裂的影响。方法培养V79细胞,分为对照组和39℃、40℃、41℃、42℃热休克预处理组,预处理后,恢复6 h,再以43℃与45℃进行高温损伤处理,4%多聚甲醛固定,抗γ-H2AX抗体标记,然后羊抗鼠FITC二抗孵育1 h,细胞核内DNA双链断裂斑点用激光共聚焦显微镜进行观察。结果在激光共聚焦显微镜下观察,经统计:对照组细胞核内的DNA双链断裂数量与热休克预处理组细胞核内的DNA双链断裂数量差异有统计学意义(P<0.05)。结论热休克预处理可以减少高温所致V79细胞DNA双链断裂的数量,说明V79细胞可以产生对温度的适应性反应。
Objective To study the influences of different heat shock pretreatment temperature on DNA double-strand breaks damaged by high temperature of V79 cells. The DNA double-strand breaks loci were acquired on a accurate laser-scanning confocal microscope and 7-H2AXs were incubated with anti y- H2AX antibodies. Methods V79 cells was cultured and divided into control group and groups with 39℃ , 40℃ ,41℃ ,42℃ heat shock pretreatment. After heat shock pretreatment and recover for 6 hours, cells were damaged with high temperature of 43℃ and 45℃. Fixed with 4% paraformaldehyde,cells were incubated with anti-y-H2AX antibodies overnight and subsequently labelled with goat-anti-mouse IgG conjugated with FITC for 1 hour. The DNA double strand breaks loci were acquired on a laser-scanning confocal microscope. Results Foci images were acquired on a laser-scanning confocal microscope. DNA double-strand breaks loci in control group and groups with heat shock pretreatment have significant differences （P〈0. 05）; Conclusion Heat shock pretreatment can decrease foci of DNA double-strand breaks induced by high temperature treatment. V79 cells can present adaptability to high temperature.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2012年第1期6-8,共3页
Cancer Research on Prevention and Treatment
基金
国家自然科学基金资助项目(10275083)
关键词
热休克预处理
Γ-H2AX
激光共聚焦显微镜
DNA双链断裂
Heat shock pretreatment
7-H2AX
Laser-scanning confocal microscope （LSCM）
DNA double strand breaks（DSBs）
