脂质体/DNA复合物介导HSV—TK基因在膀胱癌细胞的表达和生物活性

Gene Transfer of HSV-TK with DNA-Liposome Complexes in Human Bladder Cancer Cells: Expression and Biological Activity

目的：探讨脂质体／ＨＳＶ—ＴＫ重组 ＤＮＡ对膀胱癌细胞的转导和在细胞内的活性表达。方法：用脂质体／ＤＮＡ复合物将 ＨＳＶ－ＴＫ基因逆转录病毒重组子（ｐＬＸＳＮ－ＴＫ）导入膀胱癌细胞株 ＢＩＵ８７。携带 ＨＳＶ一ＴＫ基因的 ＢＩＵ８７细胞（ＢＩＵ８７—ＴＫ）由 Ｇ４１８筛选获得。经过 Ｓｏｕｔｈｅｒｎ杂交和细胞原位杂交检测后，观测 ＡＣＶ对 ＢＩＵ８７－ＴＫ细胞存活的影响。结果： Ｓｏｕｔｈｅｒｎ印迹杂交证实 ＨＳＶ—ＴＫ基因存在于 ＢＩＵ８７－ＴＫ细胞染色体中。细胞原位杂交检测 ＨＳＶ－ＴＫｍＲＮＡ的表达。外源ＤＮＡ的导入以及ＨＳＶ－ＴＫ和Ｎｅｏ基因表达不会改变细胞生长特性。然而，在ＡＣＶ作用下，转导ＨＳＶ－ＴＫ基因的膀胱癌细胞的存活率受到影响，ＡＣＶ可显著抑制癌细胞生长。结论：脂质体／ＴＫ重组 ＤＮＡ复合物可用于膀胱癌细胞的转基因治疗。

Objective TO investigate the transference of HSV-TK with DNA-liposome complexes and biological activity of HSV-TK gene in BIU87 cells. Methods: The recombinant retrovirus expression vector (pLXSN-TK) was transferred into human bladder cancer cell line BIU87 with DNA-liposome complexes. The BIU87-TK cells carrying neomycin resistance and HSV-TK gene were selected in the medium containing C418. After the analysis by Southern blot and technique of cell in situ hybridization, the influence of ACV on the BIU87 cells was investigated. Results: Southern blot analysis detected the HSV-TK gene existing in the genomic DNA of BIU87-TK cells. Expression of HSV-TK gene in HSV-TK gene-transduced cells was detected by the technique of cell in situ hybridization. Transudation of plasmid DNA or expression of neoresistance and HSV-TK gene did not significantly change the growth characteristics of BIU87 cells. But the survival rate of the bladder cancer cells of the transferred HSV-TK gene was influenced by ACV. The proliferation of HSV-TK+BIU87 cells was inhibited significantly by ACV. Conclusion: Gene transference of HSV-TK with DNA-liposome complexes may be utilized in gene therapy for human bladder cancer cells.