摘要
AIM： To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma （HCC） cell line HepG2 and normal liver cell line L02. METHODS： We constructed the recombinant replication-incornpetent Ad.rnda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the rnRNA expressing in cells, by ELISA was used to detect MDA-7/IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.rnda-7 was confirmed by Hoechst staining and flow cytometry assay with Annexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. RESULTS： Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RTPCR showed that the Ad.rnda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.rnda-7-infected L02 and HepG2 ceils, respectively. Ad.mda-7 infection obviously induced apoptosis （from 2.604±0.72% to 33.64±13.2%, P=0.00012） and growth suppression in HepG2 （inhibition ratio IR=68%） and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle （from 6.44% to 32.29%, P〈 0.01）, but not in L02 cells.CONCLUSION： These results confirm selectively induction of apoptosis and growth suppression by the mda-7/ IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2.
瞄准：调查在人的肝细胞在肿瘤生长和 apoptosis 上表示 MDA-7/IL-24 的复制无能的侵入人体气管粘膜的病菌向量的效果癌(HCC ) 房间线 HepG2 和正常的肝房间衬里 L02。方法：我们构造了 recombinant 复制无能的 Ad.mda-7 病毒向量并且感染了它进人的 HCC 房间线 HepG2 和正常的肝房间线 L02。RT-PCR 被执行检测在房间表示的 mRNA。被 ELISA 用来在文化上层清液检测 MDA-7/IL-24 蛋白质表示。Ad.mda-7 导致的 apoptosis 的效果被 Hoechst 与染色的 Annexin-V 和 PI 染色和流动血细胞计数试金证实。MTT 试金被用来决定 HepG2 细胞的生长抑制，并且房间周期和 hypodiploidy 分析被流动血细胞计数执行。结果：Recombinant 复制有缺陷者病毒表示 MDA-7/IL-24 成功地被构造。RT-PCR 证明 Ad.mda-7 能调停进 HepG2 和 L02 的外长的基因 MDA-7/IL-24 的表示。在上层清液的 MDA-7/IL-24 蛋白质的集中分别地在 Ad.mda-7-infected L02 和 HepG2 房间是 130 pg/mL 和 110 pg/mL。 Ad.mda-7 感染显然导致了 apoptosis (从2.60%+/-0.72%到33.6%+/-13.2%， P=0.00012 )并且在 HepG2 (抑制比率IR=68%)的生长抑制和在房间的 G2/M 阶段的特定的癌症房间类型的百分比的增加骑车(从6.44%～32.29%， P<0.01 )，然而并非在 L02 房间。结论：这些结果有选择地在人的肝细胞癌细胞线 HepG2 与复制无能的侵入人体气管粘膜的病菌向量由 mda-7/IL-24 基因证实 apoptosis 和生长抑制的正式就职。
