Regulation of p53 expression to overcome cisplatin rsistance

调节 p53 表达以克服顺铂耐药性

• 批准号: 09671479
• 负责人: TSUCHIYA Hiroyuki
• 金额: $ 2.37万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671479/
• 关键词: wild-type p53; transfection; cisplatin; caffeine; osteosarcoma; wild-type p53; human osteosarcoma; Osteosarcoma; p53; Cisplatin; Caffeine; Synergistic effect

The p53 tumor suppressor gene product is an important participant in the cellular response to DNA damage. This response results in either Gl cell cycle arrest or cell death by apoptosis. Therefore, the status of p53 can be expected. to affect the cellular response to DNA-damaging cytotoxic agents. The present study was performed to investigate whether introduction of the wild-type p53 gene into human osteosarcoma cells could change growth rates and enhance the cytocidal effect of cisplatin and the synergistic antitumor effect of caffeine. Three human osteosarcoma cell lines, OST (wild p53 alleles) , Saos2 (both p53 alleles deleted) , and HOS, (both mutant p53 alleles) were used. Wild-type p53 expression plasmid was transfected into each of the osteosarcoma cell lines by using the lipofection method. The transfected cells, Saos2/p53 and HOS/p53, showed a reduction in growth rate compared with the parent cells. WST-1 assay, performed to assess the cytocidal effect of cisplatin and the sy … More nergistic antitumor effect of caffeine, showed that Saos2/p53 cells were twice as sensitive to cisplatin alone in 1C50 than were Saos2 cells. The synergistic antitumor effect of caffeine also enhanced in the Saos2/p53 cell line. The other two transfected cells revealed no significant differences from their respective parent cells. Furthermore, TUNEL assay used to analyze apoptotic events in Saos2 cells and Saos2/p53 cells that were treated either with cisplatin alone or with cisplatin and caffeine revealed that compared with the parent Saos2 cell line, Saos2/p53 become most sensitive to cisplatin alone and to cisplatin with caffeine. OST/p53 and HOS/p53 cell lines did not show any significant increase in sensitivity to either cisplatin alone or to cisplatin with caffeine. These results demonstrate that the cytocidal effect of cisplatin and the synergistic antitumor effects of caffeine are enhanced by introduction of the wild-type p53 gene in a human osteosarcoma cell line with deleted p53 alleles, indicating the possibility of gene therapy using the p53 gene for human osteosarcomas with abnormal p53 status. Less

p53肿瘤抑制基因产物是细胞对DNA损伤反应的重要参与者。这种反应导致Gl细胞周期阻滞或细胞凋亡死亡。因此，p53的状态是可以预期的。影响细胞对破坏dna的细胞毒性药物的反应。本研究旨在探讨将野生型p53基因导入人骨肉瘤细胞是否能改变骨肉瘤细胞的生长速度，增强顺铂的杀细胞作用和咖啡因的协同抗肿瘤作用。使用三种人骨肉瘤细胞系OST（野生p53等位基因）、Saos2 （p53等位基因均缺失）和HOS （p53等位基因均突变）。采用脂质转染法将p53野生型表达质粒转染到骨肉瘤细胞系中。转染的细胞Saos2/p53和HOS/p53与亲本细胞相比生长速率降低。WST-1实验显示，在1C50中，Saos2/p53细胞对单用顺铂的敏感性是Saos2细胞的两倍。在Saos2/p53细胞系中，咖啡因的协同抗肿瘤作用也增强。另外两个转染的细胞与它们各自的亲本细胞没有显着差异。此外，TUNEL分析了顺铂单独或顺铂加咖啡因处理的Saos2细胞和Saos2/p53细胞的凋亡事件，结果显示，与亲本Saos2细胞系相比，Saos2/p53对顺铂单独和顺铂加咖啡因最敏感。OST/p53和HOS/p53细胞系对单独顺铂或顺铂与咖啡因的敏感性均未显示出任何显著增加。这些结果表明，在p53等位基因缺失的人骨肉瘤细胞系中引入野生型p53基因后，顺铂的细胞杀伤作用和咖啡因的协同抗肿瘤作用得到增强，提示利用p53基因对p53状态异常的人骨肉瘤进行基因治疗的可能性。少

项目成果

H.Tsuchiya: "Classification in terms of expression of p53 in cisplatin enhancement of caffeine as a biochemical modulator on human osteosarcoma cell lines." Transaction books,9th international symposium on limb salvage,New York. 141 (1997)

H.Tsuchiya：“根据顺铂增强咖啡因作为人骨肉瘤细胞系生化调节剂中 p53 表达的分类。”

土屋弘行: "ヒト骨肉腫細胞株におけるp53蛋白を標的としたcaffeineによるCDDP増強効果治療." 日本癌治療学会誌. 31(8). 712 (1996)

Hiroyuki Tsuchiya：“咖啡因针对人骨肉瘤细胞系中的 p53 蛋白增强 CDDP 治疗效果。”日本癌症治疗学会杂志 31(8)。

土屋弘行: "骨肉腫に対するカフェイン併用化学療法の分子生物学的検討-カフェインによるシスプラチン増強効果機序におけるp53蛋白の役割." 臨床整形外科. 32. 31-37 (1997)

Hiroyuki Tsuchiya：“骨肉瘤咖啡因联合化疗的分子生物学研究 - p53 蛋白在咖啡因顺铂增强作用机制中的作用。”临床骨科。

土屋弘行: "野生型p53発現骨肉腫細胞株におけるcaffeineによるcisplatin増強効果機序の検討." 日本整形外科学会雑誌. 70(6). S997 (1996)

Hiroyuki Tsuchiya：“在表达野生型 p53 的骨肉瘤细胞系中研究咖啡因增强顺铂的机制。”日本骨科学会杂志 70(6)。

土屋弘行: "野生型p53発現ヒト骨肉腫細胞のにおけるcisplatin感受性の差異によるcaffeineのcisplatin増強効果の有無に関する検討." 日本整形外科学会雑誌. 70(8). S1212 (1996)

Hiroyuki Tsuchiya：“根据表达野生型 p53 的人骨肉瘤细胞的顺铂敏感性差异来检查咖啡因是否存在顺铂增强作用。”日本骨科学会杂志 70(8)。 ）