Role Of Notch 1 Signaling In The Proliferation And Die Depth Of High Purity CD133~+ Cells From Human Hepatoma SMMC 7721 Cells

Objective To study the stem cell properties of CD 133+cells from the human hepatoma SMMC 7721 cells and investigate the effect of Notch1 signaling pathway on the proliferation and apoptosis of CD 133+ cells.Methods Sorted out and detected the CD 133+ cells from SMMC 7721 cells with Flow cytometer. Soft agar colony formation and tumor formation in nude mice were used to validate the stem cell properties of CD 133+ cells. The Notch signaling system of CD 133+ cells was measured by RT-PCR.The Jagged1, an activator of Notch signaling, and the γ-secretase inhibitor(DAPT) were added into the mediums of the CD 133+ cells respectively, the control was added with PBS buffer. Then the level of each Hes-1 m RNA was measured by RT-PCR, MTT assay and Flow cytometry were used to detect the proliferation and apoptosis of CD 133+ cells.Results The CD 133+ cells were sorted out from the SMMC 7721 cells successfully. Compared to CD 133-cells,CD 133+ cells had a stronger abilities of colony formation in vitro and tumorigenic in vivo.Only the Notch1/Jagged1 expressed in the CD 133+ cells.Expression of Hes-1 m RNA in activator group and inhibitor group had significant difference compared with control group(P<0.05),the OD490 nm values also had significant difference among the three(P<0.05), and the total apoptosis rates in the activator group,control group and inhibitor group were 46.10%、13.49% and 28.40%(P<0.05) respectively.Conclusions The SMMC 7721 cells contains CD133+cells,CD133+cells has the property of stem cells,activation and inhibition the Notch 1 of CD133+ cells is feasible, The Notch1 signaling pathway plays an important role in the proliferation and apoptosis of the CD 133+ cells,it’s helpful to futher understanding the development of tumor. The research of the biological characteristics and mechanism in CD133+ cells, it can leads tonew ideas for the treatment of liver cancer.