Li Zhi-Jin, Ye Jing-Zhu, Zhan Li-Ying, Zheng Ma-Liang
Department of Hematology, 184 Hospital of PLA, Yingtan 335000, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2016 Mar;36(3):423-8.
To investigate that the role of Axin in regulating the invasion and migration ability of lymphoma cells and explore the molecular mechanisms.
The expressions of Axin, β-catenin, MMP7, and MMP9 were detected in different lymphoma cell lines by RT-PCR and Western blotting. A lymphoma cell line with low Axin expressions was transiently transfected with pCMV5-HA-Axin and pcDNA5-His-β-catenin plasmid, and the expressions of β-catenin, MMP7, and MMP9 mRNA and protein were observed. A lymphoma cell model stably overexpressing Axin was transfected with AXIN-shRNA and β-catenin-shRNA, and the changes in β-catenin, MMP7, and MMP9 cexpressions were observed. The changes in the invasion and migration abilities of this cell model were assessed following Axin knockdown.
In the lymphoma cell lines tested, the Axin expression showed a negative correlation with β-catenin, MMP7, and MMP9 expressions. In Raji cells with a low Axin expression, overexpression of Axin resulted in decreased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, and overexpression of β-catenin obviously increased MMP7 and MMP9 mRNA and protein expressions. In the cells with stable Axin overexpression, Axin knockdown caused increased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, while β-catenin knockdown caused lowered expressions of MMP7 and MMP9 and suppressed cell invasion and migration.
In lymphoma cells, Axin overexpression can decrease the expression of β-catenin, which in turn decreases the expressions of MMP7 and MMP9 to inhibit the cell invasion and migration.
研究Axin在调节淋巴瘤细胞侵袭和迁移能力中的作用并探讨其分子机制。
采用RT-PCR和蛋白质印迹法检测不同淋巴瘤细胞系中Axin、β-连环蛋白、基质金属蛋白酶7(MMP7)和基质金属蛋白酶9(MMP9)的表达。将Axin表达低的淋巴瘤细胞系用pCMV5-HA-Axin和pcDNA5-His-β-连环蛋白质粒进行瞬时转染,观察β-连环蛋白、MMP7和MMP9 mRNA及蛋白的表达。将稳定过表达Axin的淋巴瘤细胞模型用AXIN-shRNA和β-连环蛋白-shRNA进行转染,观察β-连环蛋白、MMP7和MMP9表达的变化。在敲低Axin后评估该细胞模型侵袭和迁移能力的变化。
在所检测的淋巴瘤细胞系中,Axin表达与β-连环蛋白、MMP7和MMP9表达呈负相关。在Axin表达低的Raji细胞中,Axin过表达导致β-连环蛋白、MMP7和MMP9蛋白水平表达降低,但mRNA水平未降低,而β-连环蛋白过表达明显增加MMP7和MMP9 mRNA及蛋白表达。在稳定过表达Axin的细胞中,敲低Axin导致β-连环蛋白、MMP7和MMP9蛋白水平表达增加,但mRNA水平未增加,而敲低β-连环蛋白导致MMP7和MMP9表达降低并抑制细胞侵袭和迁移。
在淋巴瘤细胞中,Axin过表达可降低β-连环蛋白的表达,进而降低MMP7和MMP9的表达以抑制细胞侵袭和迁移。