摘要
目的探讨S100A4基因在食管癌侵袭转移中的作用和可能机制。方法化学合成2对靶向S100A4的siRNA,脂质体介导转染入人食管癌EC-9706细胞,同时以转染阴性对照siRNA细胞和未转染空白细胞作为对照。转染48 h后,半定量RT-PCR检测各组细胞S100A4和基质金属蛋白酶-9(MMP-9)基因mRNA表达的变化,Western blot检测各组细胞S100A4和MMP-9基因蛋白表达的变化;转染48 h后,侵袭小室检测各组细胞穿膜细胞数的变化,划痕实验检测各组细胞迁移能力的变化。结果 RT-PCR和Western blot结果显示:与转染阴性对照siRNA的细胞和未转染的空白细胞相比,转染2对靶向S100A4的siRNA的细胞S100A4 mRNA和蛋白的表达明显降低,差异有统计学意义(P<0.05);与转染阴性对照siRNA的细胞和未转染的空白细胞相比,转染2对靶向S100A4的siRNA的细胞MMP-9 mRNA和蛋白的表达同样明显降低,差异有统计学意义(P<0.05)。Boyden侵袭小室结果显示:与转染阴性对照siRNA的细胞和未转染的空白细胞相比,转染2对靶向S100A4的siRNA的细胞穿膜细胞数明显下降,差异有统计学意义(P<0.05);划痕实验结果显示:转染2对靶向S100A4的siRNA的细胞的迁移距离明显小于转染阴性siRNA的细胞和未转染的空白细胞,差异有统计学意义(P<0.05)。结论 S100A4参与食管癌侵袭转移的发生,S100A4可能通过影响MMP-9的表达参与此作用。
Objective To investigate the role of S100A4 in the invasion and metastasis of esophageal cancer and the possible mechanism. Methods Two siRNAs targeting $100A4 were chemically synthesized and transfected into EC - 9706 cells by LipofectamineTM 2000. EC - 9706 cells transfected with negative control siRNA and blank EC - 9706 cells were used as controls. The mRNA and protein levels of both $100A4 and MMP- 9 were evaluated by semi -quantitative RT - PCR and Western blot, respectively. Boyden chamber was used to evaluate the invasion capability of EC - 9706 cells in vitro. The mobility of EC - 9706 cells was evaluated by wound - healing assay in vitro. Results The expression of S100A4 mRNA and protein was significantly down - regulated in EC - 9706 cells transfected with 2 siRNAs targeting S100A4 （ P 〈 0. 05 ） ; so was expression of MMP - 9 mRNA and protein （ P 〈 0. 05 ） ; so were the reduction of invasion capacity mobility according to Boyden chamber resuhs and wound - healing assay （ P 〈 0. 05 ）. Conclusion S100A4 partic- ipates in the invasion and metastasis of esophageal cancer, which may act via influencing the expression of MMP- 9.
出处
《广东医学》
CAS
CSCD
北大核心
2013年第6期831-834,共4页
Guangdong Medical Journal
