Regulatory Effect Of SFMBT1 On HeLa Cell Proliferation And Migration Via Notch Signaling Pathway

Objective: In human cells,SFMBT1 is a majority protein with four repetitive MBT structures that play an important role in transcriptional silencing of cells and can interact with a variety of proteins to act on DNA transcriptional repressors.SFMBT1 plays an important role in maintaining cell dryness,accelerating cell growth,promoting cancer cell EMT and invasiveness through its transcriptional inhibition,and can promote cell progression and metastasis,which is associated with poor tumor prognosis.The aim of this study was to investigate the effects of SFMBT1 expression on the proliferation and migration of cervical cancer HeLa cells,as well as the changes in the expression levels of key factors and downstream target genes in Notch signaling pathway,and to explore the effect of SFMBT1 on Notch signaling pathway in HeLa cells.Methods: The recombinant eukaryotic expression vector prvkm-SFMBT1 containing SFMBT1 gene was constructed and identified by double enzyme digestion and gene sequencing.The SFMBT1 interference sequence of siRNA was synthesized.The blank control group,prvkm-vector group and SFMBT1-siNC group were also set.HeLa cells were transfected with prvkm-SFMBT1,prvkm-vector,siRNA-SFMBT1 and SFMBT1-siNC,respectively.After 48 h,the relative expression levels of SFMBT1 mRNA and protein were detected by RT-qPCR and Western blotting to confirm the overexpression or knockdown of SFMBT1.The expression of SFMBT1 mRNA and protein was detected by MTT method at 0h after transfection.Cell proliferation was detected at 24 h,48h,72h;after cell transfection,scratches were made in the culture dish,and the width of the scratches were measured at different times to determine the migration of the cells;SFMBT1 was detected by RT-qPCR and Western blotting.The effects of the Notch signaling pathway key factor RBP-J and the Notch3 receptor and downstream target genes HEY1,HES1 mRNA and protein expression levels;CHIP andCOIP experiments were used to detect the interaction between SFMBT1 and RBP-J.Results: Overexpression of SFMBT1 promoted the proliferation and migration of cervical cancer HeLa cells,while interference with SFMBT1 expression inhibited the proliferation and migration of cervical cancer HeLa cells.Overexpression of SFMBT1 inhibited the key factors of Notch signaling pathway RBP-J and Notch3 receptors and downstream target genes.The expression of HEY1 and HES1 mRNA and protein,while interfering with SFMBT1,played the opposite role;CHIP and COIP experiments showed an interaction between SFMBT1 and RBP-J.Conclusion: Down-regulation of SFMBT1 can significantly inhibit the proliferation and migration of HeLa cells,whereas up-regulation of SFMBT1 is the opposite;SFMBT1 can interact with RBP-J to inhibit the activation of Notch signaling pathway,which can act as a tumor-promoting factor in the specific cellular environment of cervical cancer,SFMBT1 Induction of target gene suppression by Notch signaling pathway may be a target for the treatment of cervical cancer.