Active mechanism of the surface marker CR-1 expression on the esophageal cancer stem cell growth

Objective:To investigate the active mechanism of the surface marker Cripto-1(CR-1) expression on esophageal cancer stem cell growth.Methods:Eca109 stem cells was isolated from Eca109 cells.The total proteins of HEEC,Eca109 cells and Eca109 stem cells were extracted,and the expression level of CR-1 was detected using Western-blot.The silencing CNE2/CR1^- vector targeting CR-1 gene and overexpression vector CNE1/CR-1⁺ were constructed,and transfected into the Eca109 stem cells using lipofectamine.The control group was set.The transfection efficiency was examined using RT-PCR.The proliferation of Eca109 stem cells in control group,CNE2/CR-1^- and CNE1/CR-1⁺ were detected using MTT.the prognosis of Eca109 stem cells in each group was examined using flow cytometer.The protein expression levels of Akt and p-Akt in each group were detected using Western-blot.Results:The expression levels of CR-1 in Eca109 cells and Eca109 stem cells were significantly higher than that in HEEC cells(P<0.01).RT-PCR results showed that the expression level of CR-1 in CNE2/CR-1-group was significantly lower than that in control group,and the expression level of CR-1 in CNE1/CR-1⁺ group was significantly higher than that in control group(P<0.01).MTT assay results showed that the cell proliferation in CNE2/CR-1^- group was significantly lower than that in control group,and which in CNE1/CR-1⁺ group was significantly higher than that in control group(P<0.01).Flow cytometry results showed that the apoptosis rate of cells in CNE2/CR-1^- group was significantly higher than that in control group,which in CNE1/CR-1⁺ group was significantly lower than that in control group(P<0.01).Western-blot results showed that the expression level of p-Akt protein in CNE2/CR-1^- was significantly lower than that in control group,and the expression level of p-Akt protein in CNE1/CR-1⁺ group was significantly higher than that tin control group(P<0.01).The expression change of Akt protein was not obvious(P>0.05).Conclusions:The CR-1,an esophageal cancer stem cell marker,may regulate the proliferation and apoptosis of esophageal stem cell by PI3K/Akt signaling pathway.