SRSF10 Affect Colorectal Cancer Progression By Aberrant Alternative Splicing Of RFC5

Research backgroundAccording to the survey,colorectal cancer(CRC),which has a high mortality and morbidity worldwide,is one of the top three tumors in the world.Colorectal cancer is not easy to be found in the early stage.When patients have symptoms,it often has been transferred to other organs of the body,which is one of the important causes of malignant tumor-related death.The basic pathogenesis of CRC shows that alternative splicing(AS)is closely related to the occurrence and development of CRC.The role of serine / arginine-rich splicing factor 10 in colorectal cancer(CRC)has not been fully studied.In this study,we mainly found that SRSF10 can produce different splicing variants by alternative splicing of RFC5 precursor mRNA,thus affecting the pathogenic potential of colorectal cancer.Purpose of research1.To study the expression level of SRSF10 in colorectal cancer.2.To study the effect of knockdown and overexpression of SRSF10 gene expression on the function of colorectal cancer cells.3.To study the selective splicing of RFC5 gene and the expression of its splicing variants in sh SRSF10 colorectal cancer cells.Materials and MethodsThe expression of SRSF10 gene in CRC patients was predicted by bioinformatics.SW480 and HCT116 cells were transfected with sh-SRSF10 and SRSF10 puro.The knockdown and overexpression efficiency were verified by q-PCR and Western Blot.Cell viability and proliferation were measured by CCK8,cell invasion and migration were measured by transwell,cell migration was detected by scratch test,and cell cycle and apoptosis were detected by flow cytometry.The PRIdictor website was used to predict the interaction site of SRSF10 with RFC5.Different transcripts of RFC5 pre-mRNA induced by SRSF10 were determined by agarose gel electrophoresis.ResultsThrough TCGA database analysis,the study found that SRSF10 is highly expressed in CRC patients,because the specific pathogenesis of SRSF10 in CRC remains to be fully elucidated.In this study,SW480 and HCT116 cells were transfected with lentivirus to inhibit and promote the expression of SRSF10.The effects of SRSF10 knockdown and high expression on the proliferation,invasion,migration,apoptosis and cycle of CRC cells were evaluated by CCK8,Western Blot,transwell,scratch test and flow cytometry.The results showed that SRSF10-shRNA effectively knocked down the expression of SRSF10,and SRSF10 knockdown inhibited the proliferation and migration of CRC cells,promoted apoptosis,and changed the DNA replication cycle of CRC cells.When SRSF10 was highly expressed,it promoted the proliferation and migration of CRC cells,and the cell cycle of colorectal cancer also changed.Next,this study sequenced the CRC cells with SRSF10 knockdown and predicted the downstream gene replication factor C subunit 5(RFC5),which also confirmed that SRSF10 increased the transcription variant of RFC5 exon2-AS1(S)by skipping AS1 of RFC5 exon2 to promote the development of colorectal cancer.ConclusionsThis study shows that SRSF10 knockdown can inhibit the proliferation,invasion and migration of colorectal cancer cells in vitro,while promoting colorectal cell apoptosis and changing its cycle.And found that SRSF10 knockdown can affect the AS1 jump of RFC5 exon 2 in colorectal cancer.Therefore,SRSF10 may promote the progression of colorectal cancer by producing AS1 skip isomers of RFC5 exon 2by abnormal splicing of RFC5.It may become an important target for clinical diagnosis and treatment of CRC.