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.
出处
《中华肝脏病杂志》
CAS
CSCD
2001年第4期229-231,共3页
Chinese Journal of Hepatology
