MACC1 facilitate cancer stem celllike properties of colorectal cancer cells through the PI3K/AKT signaling pathway
Objective To determine the effects and mechanism of metastasisassociated colon cancer-1(MACC1)on the biological behavior of cancer stem cell(CSCs)-like properties of colorectal cancer(CRC).Methods HCT116 cells were transfected with MACC1 overexpression vector(LV-MACC1)and MACC1 empty vector(LV-Ctrl).The transfection effect was observed under fluorescence microscope.Colony-forming assay,Transwell invasion experiment,tumor sphere formation assay were used to measure cell colony formation,invasion abilities and tumor formation capacity of cancer cells,respectively.Western blot test was used to detect the expression difference of protein level of MACC1,CD4,CD133,protein kinase B(AKT)and phosphorylated AKT(p-AKT).740 Y-P was used to determine the possible roles of PI3K/AKT in MACC1induced stem cell properties.Results 48 h after infection,more than 90%of the cells showed green fluorescence.The relative expression of MACC1 protein of LV-MACC1 was(1.03±0.04),which was higher than(0.14±0.03)of LV-Ctrl,and the difference was statistically significant(P<0.05).In the colony-forming assay,the number of colonies of LV-MACC1 was(118.70±6.12),which was more than(27.00±4.16 of LV-Ctrl),and the difference was statistically significant(P<0.05).The number of LV-MACC1 cells infested with perforations was(66.80±3.85),which was more than(27.80±1.99)of LV-Ctrl,and the difference was statistically significant(P<0.05).The numbers of tumor spheres in LV-Ctrl and LV-MACC1 were(43.8±2.1)and(63.8±2.5),and the statistics showed that the number of tumor spheres in LV-MACC1 was significantly higher than that in LV-Ctrl(P<0.05).The relative expression of CD44,CD133 and p-AKT proteins in LV-MACC1 was higher than that in LV-Ctrl(P<0.05),and there was no difference in total AKT protein expression between LV-Ctrl and LV-MACC1(P>0.05).After activation of PI3K/AKT pathway activity with 740 Y-P,the relative expression of CD44 and CD133 was significantly increased compared with that without 740 Y-P,and the difference was statistically significant(P<0.05).Conclusion MACC1 facilitates cancer stem celllike properties and malignant biological behavior in CRC cells through the PI3K/AKT signaling pathway.
Colorectal cancerCancer stem cellMetastasis-associated in colon cancer-1
万方数据
维普
