腺病毒介导的人野生型p53、GM-CSF和B7-1基因转移诱导肝癌细胞的生长抑制和凋亡  

Adenovirus-mediated transfer of human wild-type p53, GM-CSF, and B7-1 genes efficiently produces growth suppression and apoptosis of hepatocellular cells in vitro

作　　者：施明[1] 王福生[1] 刘明旭[1] 张冰[1] 邱兆华[2] 雷周云[1] Masayoshi Namha 高兰兴[4] 吴祖泽[2] 

机构地区：[1]解放军第三0二医院生物工程研究室,北京100039 [2]军事医学科学院放射医学研究所 [3]日本Okayama大学 [4]军事医学科学卫生学环境医学研究所

出　　处：《中华肝脏病杂志》2001年第4期229-231,共3页Chinese Journal of Hepatology

摘　　要：目的观察携带人野生型ｐ５３、ＧＭ－ＣＳＦ和Ｂ７－１基因的重组腺病毒载体（ＢＢ－１０２）转染ＢＥＬ－７４０２、ＨＬＥ及ＨｕＨ－７肝癌细胞后ｐ５３基因的表达，以及诱导肝癌细胞凋亡，影响肝癌细胞的增殖。方法ＢＢ－１０２以ＭＯＩ为５０ｐｆｕ／细胞　感染肝癌细胞系ＢＥＬ－７４０２（ｐ５３基因为野生型）、ＨＬＥ及ＨｕＨ－７（ｐ５３基因为突变型）。免疫组织化学祛检测ＢＢ－１０２携带的ｐ５３基因的表达，ＴｄＴ法原位检测肝癌细胞的凋亡。结果ＢＢ－１０２携带的ｐ５３基因能在转染了 ＢＢ－１０２的肝癌细胞中高效表达。转染 ＢＢ－１０２后肝癌细胞生长明显受到抑制；染毒后第 ４－ １０ ｄ期间， ＢＥＬ－７４０２、 ＨＬＥ及 ＨｕＨ－７三株肝癌细胞的平均受抑率分别为 ５８．５％、 ８１．５％及７１．１％，其中对 ｐ５３基因突变的肝癌细胞的抑制程度要大于对ｐ５３基因为野生型肝癌细胞的抑制程度。转染ＢＢ－１０２还能诱导肝癌细胞的凋亡。结论ＢＢ－１０２通过其介导ｐ５３基因的表达抑制肝癌细胞的增殖，这为ＢＢ－１０２应用于肝癌的基因治疗提供实验依据。Objective To investigate the tumor suppressor activity of recombinant adenovirus vector expressing the human wild-type p53, GM-CSF, and B7-1 proteins (designated as BB-102) in human hepatocellular carcinoma cells (HCC) in vitro. Methods The wild-type p53 BEL-7402, mutant p53 HLE, and HuH-7 HCC cell lines were infected with BB-102 at MOI of 50 in vitro. Immunohistochemical assay was used to determine p53 expressed by BB-102. Tumor suppressor activity of the expressed p53 was identified by terminal deoxynucleotidy I transferase (TdT) assay in BB-102- infected HCC cell lines. Results p53 protein was found to express in a dose-dependent manner in BB- 102-infected HCC cell lines. The proliferation of HCC cell lines were suppressed significantly at the average rates of 58.5%, 81 .5%, and 71.1 % for BEL-7402, HLE, and HuH-7 respectively from 4 to 10 days, accompanying inducing apoptosis in BB-102-infected HCC cell lines. Conclusions Besides the expression of B7-1 and GM-CSF, BB-102 is able to express p53 protein in independent manner and exerts its anti-tumor activity, which suggests that BB-102 mad be useful for gene therapy against HCC in vivo.

关 键 词：肝细胞癌 p53基因 脱噬作用 基因治疗 细胞凋亡 GM-CSF B7-1基因 

分 类 号：R735.7[医药卫生—肿瘤]