重组腺病毒介导入野生型p53,GM-CSF和B7-1基因对肿瘤化疗耐药细胞生物学行为的影响

Effect of human wild-type p53, GM-CSF and B7-1 genes on biological behavior of chemotherapy - resistant tumor cells via recombinant adenovirus vector

目的：探讨重组腺病毒分导入野生型ｐ５３，ＧＭ－ＣＳＦ和Ｂ７－１基因对肿瘤化疗耐药细胞生物学行为的影响，为临床使用该重组腺病毒治疗多药耐药的肿瘤提供实验基础。方法：选用ｐ５３异常的ＫＢ－ｓ（ＶＣＲ敏感）和ＫＢ－ｖ２００（ＶＣＲ耐药）细胞作为肿瘤化疗药物敏感和耐药的模式细胞，感染携带人野生型ｐ５３，ＧＭ－ＣＳＦ和Ｂ７－１基因的重组腺病毒后，观察这两种转基因癌细胞的生物学行为（包括药物敏感性）的变化。结果：药物敏感和耐药细胞都对腺病毒易感，重组腺病毒携带的三个外源基因（ｐ５３、ＧＭ－ＣＳＦ、Ｂ７－１基因）在两种细胞中都能得到有效表达，并且能抑制它们的生长及诱导其凋亡。耐药细胞 ＫＢ－ｖ２００在转染该重组腺病毒 ４８ｈ后，其细胞膜上 Ｐｇｐ糖蛋白的泵出药物的功能却受到显著影响，表现为罗丹明１２３在其细胞内的积累增加。ＭＴＦ的实验结果也显示出其对ＶＣＲ药物敏感性的增加，裸鼠体内实验证实了感染上述重组腺病毒的ＫＢ－ｖ２００细胞的致瘤性降低，同时能增加对化疗药物ＶＣＲ的敏感性。结论：实验研究结果提示，使用该重组腺病毒并结合化疗药物，对临床治疗多药耐药的肿瘤可能会更有效。

Objective: To explore the effect on biological behavior of chemotherapy-resistant tumor cells by human wild-type p53, GM-CSF and B7-1 genes mediated via recombinant adenovirus. Methods: p53-abnormal KB-v200 (VCR resistant) and KB-s (VCR sensitive) cell lines were used as model tumor cells, which are resistant and sensitive to chemotherapeutic drugs, respectively. After infected with recombinant adenovirus carring human wild-type p53, GM-CSF and B7-1 genes, changes in biological behavior (including drug sensitivity) of these two kinds of gene-transduced cancer cells were observed. Results: Both of the cell lines were susceptible to adenovirus, all of the three exogenous genes (p53, GM-CSF and B7-1) could be effectively expressed in these cell lines. Their growth was suppressed, and apoptosis was induced. The drug-pumping-out function of Pgp glycoprotein on the cytomembrane of drug-resistant KB-v200 cells was markedly affected 48h after transfection Of the recombinant adenovirus, as revealed by increase of the amount of rhodamine 123 accumulation in the cells. The Mrs assay also indicated the reversal of their sensitivity to VCR drugs. In vivo experiment in nude mice demonstrated reduction of tumorigenicity of the KB-v200 cells or KB-s cells infected with the recombinant adenovirus, and increase of their sensitivity to VCR. Conclusion: These study results suggest that clinical application of this recombinant adenovirus carrying multiple genes in combination with chemotherapeutic agents might be more effective in treatment of tumors with MDR.