title: The targeted gene (KDRP-CD/TK) therapy of breast cancer mediated by SonoVue and ultrasound irradiation in vitro.
abstract: Suicide gene therapy has become an effective therapy for breast cancer, and ultrasound targeted microbubble destruction (UTMD) has become a popular topic in the gene therapy field. In this study, MCF-7 cells with the KDR promoter and LSl74T cells without the KDR promoter were transfected with the recombinant plasmid pEGFP-KDRP-CD/TK using UTMD. The recombinant plasmid pEGFP-KDRP-CD/TK was transfected into MCF-7 and LS174T cells successfully with no significant difference in transfection efficiency (p>0.05). By RT-PCR, the CD/TK fusion gene was shown to be expressed in MCF-7 cells but not expressed in LS174T cells. In a cytotoxicity experiment, transgenic MCF-7 cells were sensitive to the prodrugs 5-FC and GCV. When both 5-FC and GCV were administered, the rate of cellular inhibition was significantly greater than that achieved when only one of the prodrugs was administered (p<0.001). Moreover, the inhibition rates achieved administering 5-FC, GCV and both 5-FC and GCV were all significantly greater than the gene transfection rate of 21.92±3.64% (p<0.001). However, transgenic LS174T cells were not sensitive to any prodrug. These results demonstrated that UTMD is a safe, effective and targeted gene delivery system. Also, the KDR promoter can drive expression of the CD/TK double suicide gene target in MCF-7 cells, and the targeted killing effect of the KDRP-CD/TK gene on MCF-7 cells in vitro has good synergy with expression of the CD/TK fusion gene.
authors: Department of Ultrasound, The Third Xiangya Hospital of Central South University, Changsha, Hunan 410013, China.
authors: Xing-hua
authors: XH
authors: Li
authors: Ping
authors: P
authors: Zhou
authors: Li-hua
authors: LH
authors: Wang
authors: Shuang-ming
authors: SM
authors: Tian
authors: Ying
authors: Y
authors: Qian
authors: Li-rong
authors: LR
authors: Chen
authors: Ping
authors: P
authors: Zhang
mesh_terms: D000704:Analysis of Variance; D001943:Breast Neoplasms / Q000235:genetics* / Q000628:therapy*; D045744:Cell Line, Tumor; D003287:Contrast Media / Q000494:pharmacology*; D043525:Cytosine Deaminase / Q000378:metabolism; D005260:Female; D005437:Flucytosine / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D045423:Microbubbles; D010743:Phospholipids / Q000494:pharmacology*; D011355:Prodrugs / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D013459:Sulfur Hexafluoride / Q000494:pharmacology*; D013937:Thymidine Kinase / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D014465:Ultrasonics
chemical_list: D003287:Contrast Media; D010743:Phospholipids; D011355:Prodrugs; C420843:contrast agent BR1; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005437:Flucytosine; D013937:Thymidine Kinase; D043525:Cytosine Deaminase; D015774:Ganciclovir; D013459:Sulfur Hexafluoride