干扰SULT2B1下调MMP-2抑制Hepa1-6肝癌细胞的体外迁移与侵袭

Interfering SULT2B1 inhibits migration and invasion of mouse hepatocellular carcinoma Hepa1-6 cells in vitro by down-regulating MMP-2 expression

目的研究羟基类固醇硫酸基转移酶(SULT2B1)对小鼠肝癌细胞Hepa1-6体外迁移与侵袭的影响及分子机制。方法分别用SULT2B1慢病毒干扰载体和对照载体感染Hepa1-6细胞,设立干扰组和对照组。Transwell法检测细胞的侵袭与迁移能力,Real-time PCR法检测MMP-2、MMP-9和TIMP-2的mRNA水平,明胶酶谱法检测肿瘤条件培养基(TCM)中MMP-2和MMP-9的活性。结果 SULT2B1干扰组穿过未铺和铺Matrigl胶滤膜的细胞数较对照组明显减少(P<0.05),干扰SULT2B1下调MMP-2、上调TIMP-2的mRNA水平(P<0.05),干扰SULT2B1的TCM降低MMP-2的活性(P<0.05)。结论干扰SULT2B1可抑制Hepa1-6细胞的迁移与侵袭能力,其机制主要与降低MMP-2,升高TIMP-2的表达水平有关。

Objective To investigate the effect and molecular mechanism of hydroxysteroid sulfotransferase 2B1 （SULT2B1） on the migration and invasion ability of mouse hepatocellular carcinoma Hepa1-6 cells in vitro.Methods Hepa1-6 cells were divided into SULT2B1 interference group and conrol group,which infected with lentivirus-mediated SULT2B1 vector and GFP-LV vector respectively.The invasion and migration ability of treated Hepa1-6 cells were examined by Transwell assay.The mRNA levels of matrix metalloproteinase-2 （MMP-2）,MMP-9 and tissue inhibitor of metalloproteinase-2 （TIMP-2） were determined by Real-time PCR analysis.MMP-2 and MMP-9 activities in tumor conditioned medium （TCM） of Hepa1-6 cells were detected by gelatin zymography.Results The number of Hepa1-6 cells with SULT2B1 interference invaded or migrated through the membrane without and with matrigel decreased significantly as compared with the control group （P ＜ 0.05）.MMP-2 mRNA level was down regulated in Hepa1-6 cells with SULT2B1 interference,while TIMP-2 was up-regulated in the opposite way （P ＜ 0.05）.The MMP-2 activity in TCM of Hepa1-6 cells with SULT2B1 interference obviously decreased （P ＜0.05）.Conclusions Knock-down of SULT2B1 inhibits the migration and invasion ability of Hepa1-6 cells,which contributes to down-regulating MMP-2 and up-regulating TIMP-2 levels.