The Cell Cycle Promotion Function Of ILF2 In Breast Cancer Cells

Objective: The expression level of ILF2 in different breast cancer cells was detected to investigate its potential functions in breast cancer cells.The ILF2gain-of-function cell model and lose-of-function cell model were constructed,to study the function of ILF2 gene in breast cancer cells.Methods: 1.The expression levels of ILF2 in MDA-MB231,T47 D and MCF-7 breast cancer cells were detected by Western Blot.2.PCR cloning methods was employed to construct HA-ILF2 overexpression plasmid for mamalian cells.Three of sh RNA fragments targeting ILF2 were synthesized and cloned into p LV vector containing HA promoter.3.A ILF2-silencing cell model and doxycyclin-induced ILF2-overexpressing cell model were constructed by liposome transfection and puromycin screening methods,seperately.4.Cell counting methods were employed to detect cell cycle promotion functions of ILF2,in MCF7-Tet-on-HA-ILF2 and MDA-MB231-Tet-on-sh ILF2 cells.Results: 1.ILF2 was deteted to be expressed in all three types of breast cancer cells,and showed highest expression in T47 D,the medium expression in MDA-MB231,the lowest expression in MCF-7.2.PLV-HA-ILF2 plasmid was successfully constructed for overexpresion of ILF2 in MCF7 cells.An inducible expressing ILF2 cells line MCF7-Tet-on-ILF2 was obtained.The cell proliferation were promoted in Dox induced MCF7-Tet-on-ILF2 cells compared with noninduced MCF7-Tet-on-ILF2 cells.3.Three sh RNA plasmids targeting ILF2 were successfully constructed and a stable ILF2-silencing cell model（MDA-MB231-sh ILF2）was successfully constructed.The cell cycle of MDA-MB231-Tet-on-sh ILF2 cells was inhibited by silencing ILF2,compared with control cells.Conclusions: In this research,it was found that overexpression of ILF2 in breast cancer cell line MCF7-Tet-on-HA-ILF2 can promote the proliferation of breast cancer cells and silencing of ILF2 gene in MDA-MB31-Tet-on-sh ILF2 cell line can inhibit the proliferation of breast cancer cells,and ILF2 may play an important role in breast cancer development.