Splicing Factor SRSF3 Promotes The Progression Of Cervical Cancer Through Regulating DDX5

Objective: The abnormality of alternative splicing(AS)seriously affects the occurrence and development of tumors,especially the tumor mechanism involving abnormal expression of splicing factors,which has been widely concerned in the medical field.The serine/arginine rich splicing factor family(SR family)is an important family that regulates the alternative splicing of precursor mRNA.It has been proved that serine/arginine rich splicing factor 3(SRSF3)is a key oncogene for inducing many tumors such as liver cancer,gastric cancer,rectal cancer,etc.,but the role and mechanism of SRSF3 in cervical cancer remain unclear.In order to clarify the mechanism and regulation process of SRSF3 in cervical cancer,we conducted in-depth research here.Methods:SRSF3 and DDX5 long type(DDX5-L)/DDX5 short type(DDX5-S)stable expression plasmids were constructed.Through cell culture,HeLa cells with stable overexpression of SRSF3 and DDX5-L / DDX5-S were constructed.Cell count kit-8(CCK-8),cell clone formation experiment was performed to analyze the cell proliferation ability.The ability of cell migration and invasion was analyzed by wound healing,transwell migration and cell invasion test.The effect of DDX5-L/DDX5-S on AKT transcription was studied by fluorescence quantitative PCR.The expression correlation between SRSF3,DDX5-L/DDX5-S and AKT was also studied by bioinformatics analysis.Results:First,we analyzed the high expression of SRSF3 in cervical cancer tissue samples and cancer cell lines,suggesting that SRSF3 is a gene that promotes cancer.Further theoretical results indicate that stable expression of SRSF3 promotes the proliferation,invasion and migration of HeLa cells,while knocking down SRSF3 inhibits these processes.Second,SRSF3,as a splicing factor,can regulate the alternative splicing of DDX5 exon 12.We found that knockdown of SRSF3 resulted in the production of DDX5-S mRNA,and overexpression of SRSF3 reduced the splicing percentage in(PSI)of DDX5-S.Interestingly,the overexpression of DDX5 reduces the stability of DDX5-S.In order to study the effects of DDX5-L and DDX5-S on HeLa cells,we demonstrated that DDX5-L promotes HeLa cell proliferation,migration and invasion,while DDX5-S inhibits HeLa cell proliferation,migration and invasion and has different functions.This further indicates that SRSF3 regulates splicing of DDX5 in cervical cancer.Finally,we continue to explore the mechanism of action of SRSF3 in cervical cancer.According to reports,DDX5 is a transcription cofactor of AKT,and AKT can be introduced into the development of cervical cancer.We found that although the DDX5-L can promote AKT transcription,DDX5-S can inhibit AKT transcription.In order to determine whether there is a relationship between SRSF3 and AKT,we found that SRSF3 can promote the expression of AKT.Conclusion:Highly expressed SRSF3 promotes the inclusion of DDX5 exon 12,increases the growth of DDX5,and activates the AKT signaling pathway in cervical cancer,thereby promoting the proliferation,invasion and migration of cervical cancer cells;while low expression SRSF3 promotes DDX5 exon 12 skipping,inhibiting the expression of DDX5 long type and promoting the increase of DDX5 short type,thereby inhibiting the AKT signaling pathway,thereby reducing the proliferation and metastatic ability of cervical cancer cells.In conclusion,the splicing factor SRSF3 is involved in the development of cervical cancer through the signal axis of SRSF3-DDX5-AKT.We believe that SRSF3 can promote the occurrence and development of cervical cancer,and it is a potential biomarker and a target for treatment and prognosis.