Ras/MAPK信号通路参与Galectin-3介导大肠癌细胞系LoVo增殖调控的研究

Ras/MAPK signaling pathway meditated by Galectin-3 participated in effects of biology behavior on colorectal cancer LoVo

目的:探讨Ras/MAPK信号通路是否参与了Galectin-3介导的大肠癌细胞系LoVo增殖的调控。方法:以未转染的及转染空质粒的LoVo细胞分别作为阴性对照组和阳性对照组,以前期实验中构建的Galectin-3shRNA序列并成功转染的LoVo细胞作为实验组,免疫印记方法分别检测Ras/MAPK信号通路中的ERK1/2及phospho-ERK1/2、MEK1及phospho-MEK1、K-Ras及PKC信号蛋白在干扰前后的表达情况。结果:ERK1及ERK2在干扰组表达水平与对照组相比明显减弱,F值分别为43.650和20.727,P<0.05。phospho-ERK1及phospho-E-RK2在干扰组表达水平与对照组相比明显减弱,F值分别为838.029和627.082,P<0.05。MEK1及phospho-MEK1、K-Ras、PKC蛋白激酶在干扰组的表达水平与对照组比较均显著减弱,F值分别为159.101、656.148、5737.496和663.855,P<0.05。两对照组比较差异无统计学意义。结论:ERK1/2及phospho-ERK1/2、MEK1及phospho-MEK1、K-Ras及PKC蛋白激酶在Galectin-3基因沉默中表达明显减弱,提示Ras/MAPK信号通路可能参与Galectin-3介导LoVo增殖的调控。

OBJECTIVE：To study whether Ras/MAPK signaling pathway is concerned with the effect of biology behavior of colorectal cancer LoVo mediated by Galectin-3.METHODS：LoVo cells transferred with empty PRNAT-U6.1 or nothing were regarded as negative or positive control groups,respectively,and the LoVo cells succeedingly constructed Galectin-3 shRNA sequence and transfered before were as experiment groups.The expressions of ERK1/2 and phospho-ERK1/2,MEK1 and phospho-MEK1,K-Ras and PKC were detected by Western-blot before and after being transferred by Galectin-3 shRNA.RESULTS：The expressions of ERK1 and ERK2 were all weaken compared with the comparative groups in colorectal cancer LoVo interfered by Galectin-3 shRNA.There were significant differences of the expression among the Galectin-3 shRNA group and the blank and negative groups（F were 43.650 and 20.727,P0.05）.The expressions of phospho-ERK1 and phospho-ERK2 were also weaken compared with the comparative groups in colorectal cancer LoVo interfered by Galectin-3 shRNA.There were significant difference of the expression among the Galectin-3 shRNA group and the blank and negative groups（F were 838.029 and 627.082,P0.05）.The expressions of MEK1 and phospho-MEK1,K-Ras and PKC were also weakened.There were significant difference of the expression among the Galectin-3 shRNA group and the blank and negative groups（F were 159.101,656.148,5 737.496 and 663.855,P0.05）.There was no significant difference between the two control groups.CONCLUSION：The expressions of ERK1/2 and phospho-ERK1/2,MEK1 and phospho-MEK1,K-Ras and PKC were distinctly weakened in colorectal cancer LoVo interfered with Glectin-3 shRNA,suggesting that Ras/MAPK signaling pathway signaling protein possibly participates in the regulation of proliferation in colorectal cancer LoVo mediated by Galectin-3.