基础医学与临床 ›› 2014, Vol. 34 ›› Issue (11): 1508-1513.

• 研究论文 • 上一篇    下一篇

干扰SULT2B1下调MMP-2抑制Hepa1-6肝癌细胞的体外迁移与侵袭

杨晓明1,李晓波2,殷莲华2   

  1. 1. 宁夏医科大学
    2. 复旦大学
  • 收稿日期:2014-03-24 修回日期:2014-05-23 出版日期:2014-11-05 发布日期:2014-11-03
  • 通讯作者: 李晓波 E-mail:xbli@fudan.edu.cn
  • 基金资助:
    宁夏医科大学特殊人才启动项目;家族性高胆固醇血症相关功能突变的组学筛查及基因敲除小鼠模型的建立

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

  • Received:2014-03-24 Revised:2014-05-23 Online:2014-11-05 Published:2014-11-03

摘要: 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。

关键词: 关键词:肝癌细胞, 羟基类固醇硫酸基转移酶SULT2B1, 迁移与侵袭, 基质金属蛋白酶

Abstract: 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 was decreased obviously (P<0.05). Conclusion Knock-down of SULT2B1 can inhibit the migration and invasion ability of Hepa1-6 cells, which role is mainly associated with down-regulating MMP-2 and up-regulating TIMP-2 levels.

Key words: hepatocellular carcinoma, hydroxysteroid Sulfotransferases 2B1, migration and invasion, matrix metalloproteinase-2