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热休克预处理对高温致中国仓鼠肺细胞(V79)细胞DNA双链断裂的影响

Influences of Heat Shock Pretreatment on DNA Double-strand Breaks Damaged by High Temperature of V79 Cells

  • 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。

     

    Abstract: 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 foci were acquired on a accurate laser-scanning confocal microscope and γ-H2AXs were incubated with anti-γ-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-γ-H2AX antibodies overnight and subsequently labelled with goat-anti-mouse IgG conjugated with FITC for 1 hour.The DNA double-strand breaks foci were acquired on a laser-scanning confocal microscope. Results Foci images were acquired on a laser-scanning confocal microscope.DNA double-strand breaks foci in control group and groups with heat shock pretreatment have significant differences (P<0.05); ConclusionHeat shock pretreatment can decrease foci of DNA double-strand breaks induced by high temperature treatment.V79 cells can present adaptability to high temperature.

     

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