Fibulin-2在肝癌中的表达及其与MMPs、VEGF的相关性研究

Expression of Fibulin-2 in hepatocellular carcinoma and its correlation with MMPs and VEGF

目的探讨原纤维蛋白2(Fibulin-2)在肝细胞肝癌(HCC)组织中的表达情况,并探究Fibulin-2对HepG2肝癌细胞的生物学变化以及对细胞中基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、血管内皮生长因子(VEGF)表达水平的影响。方法对20例HCC癌组织和癌旁组织中Fibulin-2的表达水平采用Western blot和qRT-PCR检测分析。在细胞实验中,将肝癌细胞分为7组:未经处理的Normal组,进行Fibulin-2沉默处理的Si1、Si2、Si3组及其阴性对照组(Si-NC组),进行Fibulin-2过表达处理的overexpresion-Fibulin2组及其阴性对照组(overexpresion-NC组)。使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法、细胞划痕实验、Transwell法以及流式细胞术分别检测细胞增殖、迁移、侵袭能力、凋亡率;并使用Western blot检测Fibulin-2过表达或沉默后对应的MMP-2、MMP-9及VEGF的表达情况。结果 qRT-PCR及Western blot实验结果显示,在翻译和转录水平,Fibulin-2在肝癌癌组织中的表达低于相对应的癌旁组织(P<0.05);Fibulin-2沉默组较普通肝癌组细胞中MMP-2、MMP-9及VEGF的表达水平出现相应的增高(P<0.05),肝癌细胞的迁移、侵袭、增殖能力增强以及细胞凋亡率减少(P<0.05);Fibulin-2过表达组较普通肝癌组细胞中MMP-2、MMP-9及VEGF的表达水平出现相应的降低(P<0.05),肝癌细胞的迁移、侵袭、增殖能力受抑制以及细胞凋亡增加(P<0.05)。结论 Fibulin-2在肝癌组织中的表达低于癌旁组织,过表达Fibulin-2可抑制肝癌细胞的恶性特征,沉默Fibulin-2则增强肝癌细胞进程。Fibulin-2可能通过调节MMP-2、MMP-9及VEGF的表达水平从而影响肝癌细胞的生物学特征。

Objective To investigate the expression of Fibulin-2 in hepatocellular carcinoma(HCC), and to explore the effects of Fibulin-2 on the biological changes of HepG2 hepatoma cells and the expression of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9) and vascular endothelial growth factor(VEGF).Methods The expression of Fibulin-2 in 20 cases of HCC carcinoma and paracancerous tissues was detected by Western blot and qRT-PCR.In the cell experiment,Fibulin-2 overexpression and silencing treatment were divided into common hepatocellular carcinoma group(common group),silence-NC group(Si-NC group),Fibulin-2 silencing group(Si group),overexpression-NC group and Fibulin-2 overexpression group.MTT colorimetry,cell scratch test,Transwell assay and flow cytometry were used to detect cell proliferation,migration,invasion and apoptosis rate.Western blot was used to detect the expression of MMP-2,MMP-9 and VEGF after Fibulin-2 overexpression or silencing.Results The results of qRT-PCR and WB experiments showed that the expression of Fibulin-2 in hepatocellular carcinoma tissues was significantly lower than that in corresponding paracancerous tissues at the translation and transcriptional levels(P<0.05).The expression levels of MMP-2,MMP-9 and VEGF in Si group were higher than those in common group(P<0 05).The migration,invasion and proliferation ability of HepG2 hepatoma cells were enhanced and the apoptosis rate was decreased(P<0 05).Compared with the normal group,the expression levels of MMP-2,MMP-9 and VEGF in the overexpression group were decreased,and the migration,invasion and proliferation ability of liver cancer cells were inhibited and the apoptosis was increased(P<0.05).Conclusion The expression of Fibulin-2 in HCC tissues was significantly lower than that in paracancerous tissues.Overexpression of Fibulin-2 could inhibit the malignant characteristics of HCC cells,while silencing Fibulin-2 could enhance the progression of HCC cells.Fibulin-2 may affect the biological characteristics of hepatocellular carcinoma cells by regulating the expression levels of MMP-2,MMP-9 and VEGF.