肝素对肝癌细胞乙酰肝素酶mRNA表达和侵袭能力的影响

Effect of Heparin on heparanase mRNA expression and invasion of human hepatoma carcinoma cells in vitro

目的:探寻肝素对乙酰肝素酶(heparanase,HPA)转染的肝癌细胞侵袭能力和乙酰肝素酶mR-NA表达的影响。方法:用含人全长乙酰肝素酶cD-NA的pcDNA3-HPA质粒转染人肝癌细胞株BEL-7404,加入不同剂量肝素进行细胞培养,用侵袭小室观察肝癌细胞穿膜细胞数,用RT-PCR法检测细胞乙酰肝素酶mRNA表达。结果:BEL-7404细胞组和空载体转染细胞组乙酰肝素酶mRNA表达均为阴性,pcDNA3-HPA质粒转染组乙酰肝素酶表达阳性,穿膜细胞数(27.00±4.30)显著高于另外2组(分别为11.67±2.52和9.67±2.89,P<0.01);加入肝素后BEL-7404细胞组和空载体转染细胞组穿膜能力和乙酰肝素酶表达均无显著变化;而pcDNA3-HPA质粒转染组穿膜细胞数随着肝素剂量的加大而逐渐减少,超过一定剂量后则不再显著减少,但该组乙酰肝素酶表达始终阳性。结论:在一定剂量范围内,肝素可抑制表达乙酰肝素酶之肝癌细胞的侵袭能力,而不影响乙酰肝素酶mRNA表达。

AIM： To explore the effect of Heparin on heparanase （HPA） mRNA expression and invasion of human hepatoma carcinoma cells （HCC） in vitro. METHODS： Human liver carcinoma cells, BEL-7404, were transfected with a full-length human HPA cDNA using a pcDNA3 expression plasmid, and different dose of Heparin was added when the cells were cultured. The invasiveness of transfected BEL-7404 was measured by the matfigel invasion assays, and the HPA mRNA expression was observed by using RT-PCR. RESULTS： The HPA mRNA expression could not be detected in BEL-7404 cells （HCC group） and HCC transfected with a control pcDNA3 plasmid lacking the HPA cDNA insert （pcDNA3 group） while the HPA expression was positive in HPA-transfected HCC （pcDNA3-HPA group）. The number of membrane-permeating cells in pcDNA3-HPA group （27.00 ±4.30） was significantly more than that in the other groups （ 11.67 ± 2.52 for HCC group and 9.67 ± 2.89 for pcDNA3 group） （P 〈 0.01）. No obvious changes in HPA expression and cell invasiveness were observed in both HCC group and pcDNA3 group after Heparin was added. The membrane-permeating cells in pcDNA3-HPA group gradually decreased along with the inerease in the dose of Heparin, and the further decrease was no longer found when added Heparin exceeded a certain quantity. The HPA expression was positive all the time in pcDNA3-HPA group. CONCLUSION： Within a certain dose range, Heparin can inhabit the invasive ability of HPA gene-transfected HCC, but does not influence the HPA mRNA expression.