中华放射医学与防护杂志

王晓梅,鞠桂芝,梅树江,等.2 Gy X射线对EL-4细胞p16、CyclinD、CDK4基因转录和蛋白表达影响[J].中华放射医学与防护杂志,2003,23(5):330-333.WANG Xiaomei,JU Guizhi,MEI Shujiang,et al.Influence on gene transcription and protein expression of p16,CyclinD,CDK4 of EL-4 cell line after 2Gy X-irradiation[J].Chin J Radiol Med Prot,2003,23(5):330-333

2 Gy X射线对EL-4细胞p16、CyclinD、CDK4基因转录和蛋白表达影响

Influence on gene transcription and protein expression of p16,CyclinD,CDK4 of EL-4 cell line after 2Gy X-irradiation

投稿时间：2003-02-08

DOI：

中文关键词: 电离辐射 p16 CyclinD CDK4

英文关键词:X-rays p16 CyclinD CDK4

基金项目:国家自然科学基金资助项目(39770193)

作者	单位
王晓梅	515031 汕头, 汕头大学医学院生物教研室
鞠桂芝	吉林大学公共卫生学院卫生部放射生物学重点实验室
梅树江	515031 汕头, 汕头大学医学院生物教研室
付士波	吉林大学公共卫生学院卫生部放射生物学重点实验室
刘树铮	吉林大学公共卫生学院卫生部放射生物学重点实验室

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中文摘要:

目的研究电离辐射对EL-4细胞p16、CyclinD、CDK4基因转录及蛋白表达的影响。探讨p16/CyclinD/CDK4负向调控通路在辐射诱导G1期阻滞中的作用。方法采用半定量RT-PCR方法检测2GyX射线照射后EL-4细胞p16mRNA水平变化,采用Northern blot方法检测zCyclinD、CDK4基因转录变化,应用免疫细胞化学方法(ICC)检测3种蛋白表达的变化。结果 2GyX射线照射后2～48hEL-4细胞p16mRNA水平增高,48h恢复至正常水平。p16蛋白表达照射后2h开始增高,12h达到峰值(P<0.01),72h恢复至正常水平。EL-4细胞周期蛋白CyclinDmRNA水平于照射后1h开始明显降低,照射后8h降至最低值。CyclinD蛋白表达于照射后8h开始明显减少,持续至照射后48h(P<0.05～P<0.01)。EL-4细胞CDK4mRNA水平于照射后1h明显降低,照射后4h～12h保持较低水平,72h恢复至正常水平。CDK4蛋白表达于照射后4h明显下降,12h降至最低,72h仍低于正常水平(P<0.01)。结论 2GyX射线照射可诱导EL-4细胞p16表达增高,CyclinD和CDK4表达降低。提示p16CyclinDCDK4负向调控通路可能在电离辐射诱导EL-4细胞G1期阻滞中发挥重要作用。

英文摘要:

Objective To investigate the effect of ionizing radiation on p16, CyclinD, CDK4 gene transcription and protein expression of EL-4 cells and to elucidate the role of p16/CyclinD/CDK4 pathways in G1 arrest. Methods RT-PCR method was employed to check p16 mRNA level. Northern blot was employed to determine CyclinD and CDK4 mRNA level. Immunocytochemstry was used to measure their protein expression. Results p16 mRNA level was dramatically increased at 2～48 h, recovered at 48h. p16 protein expression was significantly elevated at 12 h (P<0.01). CyclinD mRNA level of EL-4 cells was dramatically declined at 1 h, reached the lowest level at 8 h.CyclinD protein expression of EL-4 cells was significantly decreased at 8h and lasted to 48 h (P<0.05～P<0.01). There was a significantly descent in CDK4 mRNA level of EL-4 cells at 1 h after 2Gy X-irradiation. CDK4 mRNA level remained the lower level during 4 h to 12 h, recovered at 72 h. CDK4 protein expression of EL-4 cells dramatically reduced at 4 h, reached the bottom at 12 h, didnot recover at 72 h ( P< 0.01) after 2Gy X-irradiation. Conclusion The increasing of p16 and the reduction of CyclinD, CDK4 gene transcription and protein expression of EL-4 cells were observed after ionizing radiation. The results suggested that p16/CyclinD/CDK4 negative pathway mat play an important role in G1 arrest of EL-4 cells induced by X-rays irradiation.

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