摘要
目的研究Krüppel样因子4(KLF4)和基质金属蛋白酶9(MMP9)在原发性肝癌中的表达情况,探讨KLF4调节MMP9对原发性肝癌细胞侵袭和迁移能力的影响。方法收集原发性肝癌手术患者癌和癌旁组织标本,通过免疫组织化学、实时荧光定量PCR和免疫印迹试验(Western blot)检测50例肝癌组织及对应癌旁组织中KLF4和MMP9的表达情况;构建重组质粒,上调肝癌细胞(HepG2细胞系)的KLF4表达,检测MMP9mRNA及蛋白水平的表达情况;转染后的HepG2细胞运用Transwell侵袭试验和划痕试验观察侵袭和迁移能力的变化。结果相比癌旁组织,KLF4在肝癌组织中的表达明显降低(P<0.05),而MMP9的表达明显增高(P<0.05)。通过构建重组质粒,上调KLF4的表达,发现MMP9的mRNA和蛋白表达均降低,并影响HepG2细胞的侵袭和迁移能力。结论在原发性肝癌中,KLF4低表达,而MMP9高表达。在肝癌细胞中上调KLF4表达可引起MMP9表达下降,进而抑制肝癌细胞的侵袭和迁移。
Objective To investigate the effects of Krtippel like factor 4 （KLF4） on matrix metalloproteinase 9 （MMP9） ex- pression in hepatocellular carcinoma（HCC）. Methods A total of 50 primary hepatoce｜lular carcinoma samples and their correspond- ing adjacent tissues specimens were collected. The expression of KLF4 and MMP9 were detected by IHC,Western blot and qRTPCR. After KLF4 gene was transfected into hepatoceUular carcinoma cell line（HepG2 cell line）, the expressions of KLF4 and MMP9 were conformed by qRT-PCR and Western blot. Migration and invasion of HepG2 cell line transfected by KLF4 were detec- ted by wound-healing assay and invasion assay. Results Compared to corresponding adjacent tissues,The expression of KLF4 was significantly lower in HCCs（P〈（0.05） ,and MMP9 expression was remarkably higher in HCCs（P〈0.05）. KLF4 over-expression inhibited the expression of MMP9 on the protein and mRNA levels. Wound-healing assay and invasion assay confirmed that KLF4 regulated cell invasion and migration through regulating MMP9 expression. Conclusion KLF4 showed low expression in HCCs,and MMP9 was overexpressed, Up-regulationof KLF4 could decrease the expression of MMP9 in HepG2 cell line, which inhibited inva- sion and migration.
出处
《重庆医学》
CAS
北大核心
2015年第22期3025-3029,共5页
Chongqing medicine
基金
江西省教育厅一般项目(GJJ14056)
