骨桥蛋白基因高表达的重组腺病毒对肝癌细胞恶性生物学行为的影响

Effects of recombinant adenovirus with high expression of osteopontin gene on malignant biological behaviors of hepatocellular carcinoma cells

目的 观察骨桥蛋白（OPN）基因高表达的重组腺病毒对肝癌细胞恶性生物学行为的影响.方法 培养原发性肝癌HepG2细胞株并分为OPN基因高表达的重组腺病毒（rAd-OPN组）和rAd-NC组转染包含无效片段的重组腺病毒（rAd-NC组）,测定细胞中OPN的表达量以及肝癌细胞迁移、侵袭,选取并测定3个肿瘤转移相关基因[基质金属蛋白酶（MMP）-2、MMP-9、尿激酶型纤溶酶原激活物（uPA）]表达.结果 腺病毒转染后12、24、36、48 h时rAd-OPN组细胞中OPN的mRNA含量为1.93±0.25、2.58±0.30、3.61 ±0.57、3.88±0.63,rAd-OPN组各时间段细胞迁移率分别为（18.20±2.76）％、（43.76±7.17）％、（62.48±9.25）％、（77.33 ±10.24）％,穿膜细胞数目分别为（25.24 ±4.26）、（32.13 ±4.97）、（40.27±6.24）、（47.95±7.25）个,明显高于rAd-NC组（P＜0.05）;腺病毒转染后48 h时,rAd-OPN组中MMP-2、MMP-9、uPA的mRNA含量分别为2.32±0.49、1.78±0.28、3.14 ±0.51,均明显高于对照组rAd-NC组,P＜0.05.结论 OPN基因高表达的重组腺病毒能够促进肝细胞癌的迁移、侵袭,上调转移相关基因（MMP-2、MMP-9、uPA）的表达.

Objective To study the effect of recombinant adenovirus with high expression of osteopontin （OPN） gene on malignant biological behaviors of hepatocellular carcinoma （HCC） cells.Methods HCC （HepG2） cells were cuhured and divided into rAd-OPN group and rad NC group.HCC cells in rAd-OPN group were transfected with recombinant adenovirus with high expression of OPN gene,and thsose in NC group were transfected with recombinant adenovirus containing invalid fragment.The expression of OPN,and cell migration,invasion and metastasis associated genes was detected.Results At 12,24,36,and 48 h after adenovirus transfection,mRNA expression of OPN in rAd-OPN group was 1.93 ±0.25,2.58 ±0.30,3.61 ±0.57 and 3.88 ±0.63,cell migration rate was （18.20 ±2.76）%,（43.76 ±7.17）%,（62.48 ±9.25）% and （77.33 ± 10.24）%,and transmembrane cell number was （25.24 ±4.26）,（32.13 ± 4.97）,（40.27 ± 6.24） and （47.95 ± 7.25） cells respectively,significantly higher than in rAd-NC group （P 〈 0.05）.At 48 h after transfection of adenovirus,mRNA expression of matrix metalloproteinase （MMP）-2,MMP-9,urokinase-type plasminogen activator （uPA） in rAd-OPN group was respectively 2.32 ±0.49,1.78 ±0.28,3.14 ±0.51,significantly higher than that in rAd NC group （P 〈 0.05）.Conclusion Recombinant adenovirus with high expression of OPN gene can promote the migration,invasion and up-regulate the metastasis-related gene expression.