Knockdown of MACC1 expression suppressed hepatocellular carcinoma cell migration and invasion and inhibited expression of MMP2 and MMP9
Knockdown of MACC1 expression suppressed hepatocellular carcinoma cell migration and invasion and inhibited expression of MMP2 and MMP9
复制标题
MACC1表达的敲低抑制了肝癌细胞的迁移和侵袭，并抑制了MMP2和MMP9的表达
DOI：
10.1007/s11010-012-1545-y
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发表时间：
2013-04-01
影响因子：
4.3
通讯作者：
Yao, Yingmin
中科院分区：
文献类型：
--
作者：
Gao, Jie;Ding, Feihu;Yao, Yingmin
MACC 1（结肠癌转移相关蛋白-1）蛋白的表达与多种人类癌症的转移相关。本研究分析了MACC 1蛋白在肝细胞癌（HCC）组织标本中的表达，然后研究了MACC 1敲低对HCC细胞迁移和侵袭以及基因表达水平的影响。采用免疫组化方法分析60对HCC患者和癌旁正常肝组织中MACC 1的表达。分别采用qRT-PCR和western blot方法检测肝癌细胞株Hep 3B、Huh 7、MHCC 97 H、SMMC-7721、Bel-7402和HepG 2及正常肝细胞株LO 2中MACC 1 mRNA和蛋白的表达，并采用MACC 1短发夹RNA（shRNA）抑制Huh 7细胞中MACC 1蛋白的表达。用伤口愈合试验和侵袭试验分析这些细胞的肿瘤表型的变化，并通过蛋白质印迹法评估基因表达的差异。免疫荧光法检测MACC 1蛋白在上述细胞系中的表达。MACC 1蛋白在肝癌组织中高表达，其核表达与肝癌分化程度低、肝内转移或门脉浸润有关。此外，肝癌细胞株中也有MACC 1 mRNA和MACC 1蛋白的表达。免疫组化结果显示，MACC 1蛋白定位于肝癌细胞的胞核和胞浆中，且MACC 1蛋白的核定位与肝癌细胞的侵袭性有关。使用MACC 1-shRNA敲低MACC 1表达降低Huh 7细胞的迁移和侵袭能力，这与Huh 7细胞中MMP 2、MMP 9和c-Met蛋白的下调有关。MACC 1蛋白定位于细胞核可预测HCC进展。利用MACC 1 shRNA敲低MACC 1表达作为控制HCC的新治疗策略值得进一步评价。
Expression of MACC1 (metastasis-associated in colon cancer-1) protein is associated with metastasis of various human cancers. This study analyzed MACC1 protein expression in hepatocellular carcinoma (HCC) tissue specimens and then investigated the effects of MACC1 knockdown on HCC cell migration and invasion, and gene expression levels. Sixty pairs of HCC and adjacent normal liver tissues from HCC patients were analyzed for MACC1 expression immunohistochemically. The HCC cell lines Hep3B, Huh7, MHCC97H, SMMC-7721, Bel-7402, and HepG2 and the normal liver cell line LO2 were used to assess expressions ofMACC1mRNA and MACC1 protein using qRT-PCR and western blot, respectively.MACC1short hairpin RNA (shRNA) was used to knockdown MACC1 protein expression in Huh7 cells. Changes in the tumor phenotype of these cells were analyzed with wound healing assay and invasion assays, and differences in gene expression were evaluated via western blot. Immunofluorescence was used to locate MACC1 protein in the above cell lines. MACC1 was highly expressed in HCC tissues and the nuclear expression of MACC1 protein was associated with poor tumor differentiation and intrahepatic metastasis or portal invasion. Moreover,MACC1mRNA and MACC1 protein was also expressed in HCC cell lines. Immunostaining showed that MACC1 protein was localized in both nuclei and cytoplasm of HCC cell lines and the nuclear localization of MACC1 protein was associated with increased aggressiveness of HCC in cell lines. Knockdown ofMACC1expression usingMACC1-shRNA reduced Huh7 cell migration and invasion abilities, which was associated with downregulation of MMP2, MMP9, and c-Met proteins in Huh7 cells. Localization of MACC1 protein to the nucleus may predict HCC progression. Knockdown of MACC1 expression usingMACC1shRNA warrants further evaluation as a novel therapeutic strategy for control of HCC.