Cytotoxic gene therapy for human breast cancer in vitro.
Cytotoxic gene therapy for human breast cancer in vitro.
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DOI：
10.1016/j.jss.2006.05.021
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发表时间：
2006-11
期刊：
影响因子：
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通讯作者：
S. Levy;B. Zhou;N. Ballian;Zhijun Li;Shihe Liu;Mark A. Feanny;Xiao-ping Wang;D. Blanchard;F. Brunicardi
中科院分区：
文献类型：
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作者：
S. Levy;B. Zhou;N. Ballian;Zhijun Li;Shihe Liu;Mark A. Feanny;Xiao-ping Wang;D. Blanchard;F. Brunicardi
背景转录因子PDX-1在人胰腺癌和乳腺癌中均有表达。虽然PDX-1导向的RIP-TK/GCV基因治疗对胰腺癌细胞的细胞毒性已被证实，但这种治疗在乳腺癌细胞中的疗效尚不清楚。本研究旨在探讨PDX-1在人乳腺癌细胞AU565和T47D中的表达及其对RIP活化的影响。我们还观察了RIP-TK/GCV基因治疗的效果，并检测了外源性PDX-1TO是否会增强其细胞毒作用。用RSVLacZ和RIPLacZ基因构建进行瞬时转染，用报告基因检测LacZ的表达。用腺病毒载体转染腺病毒T47D细胞。将RIP-TK基因导入细胞，确定每株细胞的次适GCV水平。GCV治疗后，用MTS法检测细胞毒作用。检测外源性PDX-1对LacZ表达和RIP-TK细胞毒作用的影响。外源性PDX-1促进AU565细胞LacZ表达，但对T47D细胞LacZ表达无明显影响。腺病毒转染法在T47D细胞中的转染率高于非病毒转染法。RIP-TK对AU565和T47D细胞具有细胞毒作用，外源性PDX-1对AU565和T47D细胞的细胞毒作用均增强。结论SRIP-TK/GCV对人乳腺癌细胞具有细胞毒作用，外源性PDX-1增强细胞毒作用。有必要进行体内研究，以确定这种治疗方法的肿瘤特异性和疗效。
BACKGROUNDTranscription factor PDX-1 is expressed by human pancreatic and breast cancers. Although cytotoxicity of PDX-1-directed RIP-TK/GCV gene therapy to pancreatic cancer cells has been demonstrated, the efficacy of this treatment in breast cancer cells is unknown. The purpose of this study was to determine the expression of PDX-1 and its effect on RIP activation in two human breast cancer cell lines, AU565 and T47D. We also investigated the efficacy of RIP-TK/GCV gene therapy and examined whether exogenous PDX-1 to would enhance its cytotoxic effect.MATERIALS AND METHODSRT-PCR was used to determine PDX-1 expression. Gene constructs RSVLacZ and RIPLacZ were used for transient transfection and LacZ expression was determined using reporter assays. T47D cells were also transfected with adenoviral vectors. Cells were transfected with RIP-TK and the suboptimal level of GCV was determined for each cell line. Following GCV treatment, cytotoxicity was measured using MTS assays. The effect of exogenous PDX-1 on LacZ expression and RIP-TK cytotoxicity was determined.RESULTSPDX-1 mRNA was expressed in human breast cancer cells and activated the RIP. Exogenous PDX-1 enhanced LacZ expression in AU565 cells but not in T47D cells. Adenoviral transfection was more efficient in T47D cells than non-viral transfection. RIP-TK treatment was cytotoxic to AU565 and T47D cells and this effect was enhanced by exogenous PDX-1 with both transfection methods.CONCLUSIONSRIP-TK/GCV therapy is cytotoxic to human breast cancer cells and exogenous PDX-1 enhances cytotoxicity. In vivo studies are necessary to determine the tumor specificity and efficacy of this treatment.