The Role And Mechanism Of SHCBP1 In Regulating Mitosis Of Gastric Cancer Cells

Objective: SHCBP1 is a novel mitotic regulator,and its mechanism in the occurrence and development of gastric cancer has not yet been elucidated.This study aimed to explore whether SHCBP1 can participate in the regulation of gastric cancer cell mitosis and its potential mechanism.Methods: The expression of SHCBP1 in gastric cancer cells during mitosis and the effect of knockdown of SHCBP1 on mitosis in gastric cancer cells were investigated by Western blot（WB）and immunofluorescence experiments.Immunofluorescence staining and live cell imaging were used to explore the localization of SHCBP1 in different stages of mitosis and the effect of SHCBP1 knockdown on spindle morphology.Finally,a gastric cancer cell line stably expressing GFP-SHCBP1 fusion protein was constructed to explore the dynamic localization of SHCBP1 in gastric cancer cell mitosis.Results: After synchronization of the cell cycle,Western blot and immunofluorescence experiments showed that SHCBP1 was significantly overexpressed in mitosis.Flow cytometry showed that knockdown of SHCBP1 gene in gastric cancer cells caused cell arrest in G2/M phase.Spindle immunofluorescence staining and live cell imaging showed that SHCBP1 was localized to the spindle at metaphase and the cytokinesis ring at telophase.Knockdown of SHCBP1 inhibits cell division and forms multipolar and defective spindles.Finally,high-content detection of GFP-SHCBP1 fusion protein was used to find the dynamic localization of SHCBP1 in mitosis.SHCBP1 could localize to punctate organelles after mitosis,and immunofluorescence staining showed that the localized punctate organelles were midbody remnant.Conclusion: SHCBP1 regulates mitosis and promotes gastric cancer growth by participating in the formation of spindle,midbody and midbody remnants,which is worthy of further study.