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

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）。免疫组织化学法检测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4－10d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。

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

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.