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中华乳腺病杂志(电子版) ›› 2008, Vol. 02 ›› Issue (01) : 40 -43. doi: 10.3877/cma.j.issn.1674-0807.2008.01.012

实验研究

突变p53 基因的反义拯救研究
孙玉兰1, 王亚红1, 张媛媛1, 许少峰1, 付丽1,()   
  1. 1.300060 天津,天津医科大学附属肿瘤医院乳腺病理研究室教育部乳腺癌防治重点实验室
  • 收稿日期:2007-12-03 出版日期:2008-02-01
  • 通信作者: 付丽
  • 基金资助:
    天津市自然科学基金基础研究重点项目(033801511)

Research of antisense rescue of mutant p53 gene in vitro

Yu-lan SUN1, Ya-hong WANG1, Yuanyuan ZHANG1, Shao-feng XU1, Li FU,1()   

  1. 1.Department of Breast Pathology and Research,Cancer Hospital,Tianjin Medical University;Key Laboratory of Breast Cancer Prevention and Therapy, Ministry of Education, Tianjin 300060,China
  • Received:2007-12-03 Published:2008-02-01
  • Corresponding author: Li FU
引用本文:

孙玉兰, 王亚红, 张媛媛, 许少峰, 付丽. 突变p53 基因的反义拯救研究[J/OL]. 中华乳腺病杂志(电子版), 2008, 02(01): 40-43.

Yu-lan SUN, Ya-hong WANG, Yuanyuan ZHANG, Shao-feng XU, Li FU. Research of antisense rescue of mutant p53 gene in vitro[J/OL]. Chinese Journal of Breast Disease(Electronic Edition), 2008, 02(01): 40-43.

目的

研究p53 基因的个体性反义RNA 联合野生型p53(wt-p53)对乳腺癌细胞的增殖抑制作用,探讨体外双重基因治疗的意义。

方法

将wt-p53 重组质粒pC53-SN3 转染人乳腺癌MDAMB-231 细胞,G418 筛选稳定表达wt-p53 的细胞克隆(WTp53-231 细胞);体外构建针对MDA-MB-231 细胞p53 基因突变外显子8(exon8)的反义表达载体[pGEM3zf( + /-)p53exon8],并制备其反义RNA(ASp53exon8'RNA);以MDA-MB-231 细胞为对照,阳离子脂质体介导下ASp53exon8'RNA 转染WTp53-231 细胞;转染48 h 后,用MTT 法检测细胞生长活性,TUNEL 法检测细胞凋亡情况,免疫细胞化学LSAB染色法检测p53 蛋白的表达。

结果

ASp53exon8'RNA 转染WTp53-231 细胞与ASp53exon8'RNA 转染MDA-MB-231 细胞比较,前者可进一步抑制肿瘤细胞增殖和诱导凋亡。

结论

p53 基因的个体性反义RNA 联合wt-p53 共转染可协同抑制乳腺癌细胞增殖,达到“反义拯救”基因治疗目的。

Objective

To investigate the inhibitive efficacy of individual antisense RNA in combination with wild-type p53 gene(wt-p53) in vitro on the proliferation of breast cancer cells and explore its possible gene therapeutic implications.

Methods

pC53-SN3 plasmids were transferred into MDA-MB-231 cells and transfected WTp53-231 cells which stably expressed wt-p53 were screened with G418. MDA-MB-231 cells and Wtp53-231 cells were exposed to cationic liposome-ASp53exon8'RNA,and the growth inhibition rate,expression of p53,and induction of apoptosis were measured with MTT method,TUNE method,and immunocytochemistry staining with LSAB method respectively 48 hours later.

Results

When WTp53-231 cell was transfected with ASp53exon8'RNA,the growth inhibition of tumor cells and increase of apoptotic rate were manifested in WTp53-231 cell, furthermore,synergistic effects were observed compared with MDA-MB-231 cell.

Conclusions

ASp53exon8'RNA in combination with wt-p53 gene transfer can synergistically suppress cell growth,which may serve as effective strategy for human cancer treatment.

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