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目的:探讨载脂蛋白A-I结合蛋白(AIBP)在肝癌细胞中的表达及意义。方法:用RT-PCR与Western blot法分别检测正常肝细胞株L02,AFP阳性人肝癌细胞株HepG2和Hep3B,及AFP阴性人肝癌细胞株SMMC7721中AIBP基因与蛋白的表达;构建AFP启动子驱动的双自杀基因(CD,TK)+AIBP基因过表达载体pcDNA3.1-AFP-AIBP-yCD/TK,并转染Hep3B和SMMC7721细胞,用MTT法检测转染后细胞的增殖能力,用RT-PCR和Western blot法检测细胞AIBP,血管内皮生长因子(VEGF),VEGF受体2(VEGFR-2),基质金属蛋白酶9(MMP-9)基因和蛋白的表达。结果:AIBP mRNA和蛋白在正常肝细胞中高表达,而在各肝癌细胞系中均表达下调,且Hep3B和SMMC7721细胞中下调明显。成功构建pcDNA3.1-AFP-AIBP-yCD/TK真核表达质粒并转染入Hep3B和SMMC7721细胞。转染后AFP阳性Hep3B细胞生长到明显抑制,但AFP阴性SMMC7721细胞增殖不受影响;两种细胞的AIBP基因与蛋白表达均明显上调,而VEGFR-2,VEGF和MMP-9基因与蛋白表达明显表达下调。定量指标间的差异均有统计学意义(均P<0.05)。结论:AIBP在肝癌细胞中表达下调,AIBP与肝癌细胞的侵袭转移能力有关,而与细胞增殖能力无关;成功构建了联合基因载体pcDNA3.1-AFP-AIBP-yCD/TK。
Objective: To investigate the expression of Apo-A binding protein (AIBP) in hepatocellular carcinoma cells and its significance. Methods: The expression of AIBP gene and protein in normal liver cell line L02, AFP positive human hepatoma cell line HepG2 and Hep3B, and AFP negative human hepatocellular carcinoma cell line SMMC7721 were detected by RT-PCR and Western blot, respectively. AFP promoter-driven The double suicide gene (CD, TK) + AIBP gene overexpression vector pcDNA3.1-AFP-AIBP-yCD / TK was transfected into Hep3B and SMMC7721 cells. The proliferation of transfected cells was detected by MTT assay. Western blot was used to detect the expression of AIBP, VEGF, VEGFR-2 and MMP-9. Results: AIBP mRNA and protein were highly expressed in normal liver cells, but down-regulated in all hepatocellular carcinoma cell lines, and down-regulated in Hep3B and SMMC7721 cells. The pcDNA3.1-AFP-AIBP-yCD / TK eukaryotic expression plasmid was successfully constructed and transfected into Hep3B and SMMC7721 cells. The AFP positive Hep3B cells were significantly inhibited after transfection, but the proliferation of AFP negative SMMC7721 cells was not affected. The expression of AIBP gene and protein were up-regulated in both AFP and VEGFR-2, VEGF and MMP-9 genes and proteins Down regulation. There were significant differences between the quantitative indicators (all P <0.05). CONCLUSION: The expression of AIBP is down-regulated in hepatocellular carcinoma cells. AIBP is related to invasion and metastasis of hepatocellular carcinoma cells, but not to cell proliferation ability. A combined gene vector pcDNA3.1-AFP-AIBP-yCD / TK was successfully constructed.