论文部分内容阅读
目的 :探讨HSV TK/GCV(单纯疱疹病毒Ⅰ型胸苷激酶 /丙氧鸟苷 (substrateⅠofherpessimplexvirusthymidinekinase/ ganciclovir)系统及其与普通化疗联合用于治疗卵巢癌的可行性 ,研究分子化疗药物GCV与普通化疗药多柔比星 (阿霉素 )联合应用对卵巢癌SK OV 3/TK细胞的杀伤作用。这对于缺乏有效的早期诊断和治疗手段的卵巢癌具有特殊的意义。方法 :将携带HSV TK基因的逆转录病毒重组体 (XM 6 /TK)以磷酸钙共沉淀法导入包装细胞 ,经G4 18筛选获得产病毒的阳性克隆细胞。继而以包装好的复制缺陷型逆转录病毒为运载体将HSV TK基因导入SK OV 3卵巢癌细胞。再经G4 18筛选 ,确定目的基因在靶细胞内稳定表达后 ,观察GCV对细胞的杀伤作用。进一步观察GCV与多柔比星联合应用对SK OV 3/TK细胞的杀伤作用。结果 :HSV TK基因可由重组逆转录病毒导入卵巢癌细胞并能获得稳定表达 ;GCV(10mg·L-1)可明显抑制被转化的卵巢癌细胞 (SK OV 3/TK)的生长 ,对于SK OV 3/TK细胞 ,在单独应用GCV时 ,其最低有效 (质量 )浓度为 10mg·L-1;单独应用多柔比星时 ,其最低有效 (质量 )浓度为 1mg·L-1;而联合应用时 ,GCV 5mg·L-1、多柔比星 0 .2 5mg·L-1对SK OV 3/TK细胞即可产生明显的杀伤作用。结论 :逆转录病毒介导的HSV TK/GCV系统可
Objective: To investigate the feasibility of HSV TK/GCV (substrate I of herpes simplex virus thymidine kinase/ganciclovir) system combined with chemotherapy in the treatment of ovarian cancer, and to study the molecular chemotherapeutic agent GCV and chemotherapy Drug Doxorubicin (Doxorubicin) Combined Use of Killing Effects on Ovarian Cancer SK OV 3/TK Cells This is of special significance for ovarian cancer that lacks effective early diagnosis and treatment.Methods: Will carry the HSV TK gene The recombinant retroviruses (XM 6 /TK) were introduced into packaging cells using a calcium phosphate co-precipitation method, and virus-producing positive clones were obtained by G4 18 screening, and then the HSVs were packaged using a replication-defective retrovirus as a carrier. The TK gene was transfected into SK OV 3 ovarian cancer cells, screened by G4 18 to determine the stable expression of the target gene in the target cells, and the killing effect of GCV on the cells was observed. Further observation of GCV combined with doxorubicin for SK OV 3/ The killing effect of TK cells.Results: HSV TK gene can be introduced into ovarian cancer cells by recombinant retrovirus and can be stably expressed; GCV (10mg·L-1) can be obviously The growth of transformed ovarian cancer cells (SK OV 3/TK) was the lowest effective (mass) concentration of SK OV 3/TK cells when GCV alone was 10 mg·L -1; Doxorubicin alone was used; The minimum effective (mass) concentration is 1 mg·L-1 when used as a star, while the combination of GCV 5 mg·L-1 and 0.25 mg·L-1 doxorubicin can be used for SK OV 3/TK cells. Significant killing effect.Conclusion: retrovirus-mediated HSV TK/GCV system can be