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目的研究骨桥蛋白(OPN)促进肝癌转移的可能机制。方法经质粒转染、G418筛选获得稳定高表达OPN人肝癌细胞株SMMC-7721,以空质粒转染组为对照。比较两组细胞的肿瘤转移相关基因表达差异。选取其中3个肿瘤转移相关基因(MMP-2、MMP-9、uPA),用ELISA方法检测其细胞培养上清液的蛋白表达情况。结果经肿瘤转移基因芯片比较发现SMMC-7721高表达OPN后,有88个肿瘤转移相关基因表达上调,有6个肿瘤转移相关基因表达下调。高表达OPN SMMC- 7721上清液MMP-2、uPA水平明显高于对照组[(46．11±3．12)vs(19．66±3．69)μg/L,(5．36±0．94)vs(1．39±0．64)μg/L,而MMP-9表达水平差异无统计学意义[(7．97±2．42)vs(7．25±2．14)μg/L,]与基因芯片结果符合。结论在SMMC-7721肝癌细胞株中OPN高表达引起多个肿瘤转移相关基因改变。OPN可能通过上调MMP-2、uPA等基因表达促进肝癌转移。
Objective To investigate the possible mechanism of osteopontin (OPN) in promoting the metastasis of liver cancer. Methods The stable and highly expressed OPN human hepatocellular carcinoma cell line SMMC-7721 was obtained by plasmid transfection and G418 screening. The empty plasmid transfection group was used as a control. Differences in tumor metastasis-related gene expression between the two groups were compared. Three of the tumor metastasis related genes (MMP-2, MMP-9, uPA) were selected and the protein expression of cell culture supernatants was detected by ELISA. Results After SMMC-7721 was overexpressed OPN, 88 tumor metastasis-associated genes were up-regulated and 6 metastasis-associated genes were down-regulated by tumor metastasis gene chips. The level of MMP-2 and uPA in the supernatant of OPN SMMC-7721 cells was significantly higher than that in the control group [(46.11 ± 3.12) vs (19.66 ± 3.69) μg / L, (5.36 ± 0 .94) vs (1.39 ± 0.64) μg / L, while there was no significant difference in the expression of MMP-9 (7.97 ± 2.42 vs 7.25 ± 2.14 μg / L,] in line with the results of the gene chip. Conclusion The overexpression of OPN in SMMC-7721 hepatocarcinoma cell line leads to several tumor metastasis related gene changes. OPN may promote the metastasis of liver cancer by up-regulating gene expression of MMP-2 and uPA.