PGL3DF3DTA特异性杀伤移植瘤裸鼠人乳腺癌细胞的实验研究
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黄文广

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Experimental study on PGL3DF3DTA specific killing of tumors from implanted human breast cancer cells in nude mice
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    摘要:

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    目的 研究新构建的含人乳腺癌DF3启动子和白喉毒素A片段的重组表达载体PGL3DF3DTA对裸鼠人乳腺癌移植瘤的特异性杀伤作用。
    方法 建立人乳腺癌移植瘤裸鼠动物模型,将4周龄裸鼠36只随机分为实验组、空载体组、空白对照组、阴性对照组4组,每组9只。将DF3阳性和阴性的人乳腺癌细胞MCF7和MDAMB231分别接种于相应的裸鼠皮下。待成瘤后,将重组表达载体PGL3DF3DTA和PGL3DF3多次、多位点注射入相应裸鼠移植瘤内。连续测量瘤体大小,计算瘤体体积。在不同时间点分批处死部分动物,行形态学观察、检测肿瘤细胞的凋亡,检测Ki67,Bax及Bcl2蛋白的表达。
    结果 裸鼠人乳腺癌动物模型建立成功,经重组表达载体PGL3DF3DTA作用后的人乳腺癌细胞出现明显的凋亡现象, Ki67及Bcl2基因表达水平降低,Bax基因表达水平升高。
    结论 重组表达载体PGL3DF3DTA能对DF3阳性的乳腺癌细胞产生特异性杀伤作用。

    Abstract:

    Abstract:Objective To study the specific killing effect of recombinant expression vector containing human breast cancer DF3 promotor and diphtheria toxin A fragment on human breast cancer cells implanted in vivo. Methods To establish human breast cancer xenograft in nude mice,thirtysix nude mice were divided into four groups at random,nine animals in each group. Human breast cancer cells of DF3 positive and negative(MCF7 and MDAMB231)were inoculated into corresponding animals respectively. Recombinant expression vector PGL3DF3DTA and PGL3DF3 were inoculated into corresponding tumor after if developed in the animals. The tumor weight was measured and calculated continually. The animals were killed at different time periods. The morphological changes were observed. The apoptosis index of cells was evaluated by insitu TUNEL. Immunocytochemical methods were used to detect the expression of Ki67,Bax and Bcl2 protein.
    Results Human breast cancer xenograft were established in nude mice successfully. The human breast cancer cells treated by PGL3DF3DTA showed a typical apoptosis morphology and TUNEL detection analysis revealed the apoptosis index of the experimental groups was significantly enhanced,which was associated with time and dosage. The expression of Ki67 and Bcl2 were downregulated, and the expression of Bax was elevated. 〖WTHZ〗Conclusions Recombinant expression vector PGL3DF3DTA could produce specific killing effect on DF3 positive human breast cancer cell line.

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蔡明,黄文广,潘晟. PGL3DF3DTA特异性杀伤移植瘤裸鼠人乳腺癌细胞的实验研究[J].中国普通外科杂志,2007,16(1):23-.
DOI:10.7659/j. issn.1005-6947.2007.01.023

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  • 收稿日期:2006-03-30
  • 最后修改日期:2006-06-29
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  • 在线发布日期: 2007-01-25