摘要
目的探讨单纯疱疹病毒胸苷激酶(HSVTK)基因在肺腺癌基因治疗中的应用。方法对转导了HSVTK基因的感染人肺腺癌细胞A549(A549/TK)和野生型A549,采用噻唑蓝(MTT)法测定不同浓度丙氧鸟苷(GCV)作用72小时后的细胞存活率,求得半抑制浓度(IC50)。动物实验将9×106的A549/TK或野生型A549细胞接种于Balb/cnu/nu裸鼠的皮下,当肿瘤长至200~300mg时,随机分组。A549/TK组(1组)荷瘤小鼠给予每天100mg/kgGCV腹腔注射,连续给药2周,对照组(2组)A549/TK给予生理盐水,野生型A549组(3组)给予等量GCV,从给药开始之日起,每周二次测量肿瘤大小,按经验公式计算肿瘤相对重量。结果体外细胞毒实验:在20μmol/LGCV存在时,野生型A549形态无明显改变,而实验组细胞明显死亡。GCV对野生型A549的IC50为500μmol/L,对A549/TK的IC50为05μmol/L,二者相差1000倍(P<0.01)。动物实验:腹腔注射GCV可使A549/TK皮下肿瘤显著缩小。从注射之日起到17天时,肿瘤体积下降了92%,而GCV对野生型A549肿瘤?
Objective Evaluate the clinical potential of HSV TK/GCV system in gene therapy of tumors by using this system to treat human lung cancer A549 cells in vitro and in vivo. Method The in vitro sensitivity levels of A549/TK (TK tranfered A549) cells and wild type A549 cells to GCV were expressed as IC 50 which was measured with MTT assay after treating both cells for 72 hrs. In vivo study, The nude mice bearing A549/TK tumor or A549 tumor were used as experimental models. All the animals were divided into 3 groups. Group 1, the animals bearing A549/TK tumor would be treated with GCV. Group 2, The animals bearing A549/TK tumor would be treated with saline. Group 3, The animals bearing A549 tumor would be treated with GCV. When the tumors reached to 200~300 mg the animals would be injected intraperitoneally with GCV solution (100 mg·kg -1 ·day -1 ) or equal volume of saline for two weeks. In this period and the following four weeks, the tumor weights were measured and calculated with the experience formula once every three or four days. Result In vitro study showed that the A549/TK cells were apparently killed in a 20 μmol/L GCV solution while the wild type A549 cells grew normally under the same condition. The IC 50 of A549/TK cells in GCV solution was 0.5 μmol/L and that of A549 cells 500 μmol/L, showing a 1 000 fold difference ( P <0.01). In vivo experiment GCV had compelled the A549/TK tumors to lead to a regression of 92%, meanwhile the A549 tumors treated with GCV increased 3.9 times in weight in the first seventeen days of experiment. Group 2 tumors A549/TK tumors which were treated with saline, grew as well as group 3 did. There was a significant difference ( P <0.01) between group 1 and other groups. Conclusion GCV could specifically and effectively kill the TK transfered A549 cells in vitro and in vivo. This result suggested that the HSV TK/GCV system may have a clinical potential for gene therapy of lung cancer.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
1998年第5期265-267,共3页
Chinese Journal of Tuberculosis and Respiratory Diseases
关键词
肺肿瘤
HSV-TK基因
基因治疗
Lung neoplasm Adenocarcinoma Cell line Tiansfection HSV TK gene Gene therapy