POTEE Drives Colorectal Cancer Development Via Regulating SPHK1/p65 Signaling

Background:Colorectal cancer is the third most commonly diagnosed cancer(10.2%of the total cases)and the second leading cause of cancer related deaths(9.2%of the total cancer deaths)in 2018 globally.Both incidence and deathrates of colorectal cancer are increasing rapidly and maintain an upward trend in Asian countries.The global burden of colorectal cancer(CRC)is expected to increase by 60%to more than 2.2 million new cases and 1.1 million deaths by 2030.Exploring related genes in the development of CRC and finding important links that affect the biological characteristics of CRC are critical ways to understand the malignancy of tumors and to improve the survival and prognosis of CRC patients.POTE(Prostate,Ovary,Testes,and Embryo)is a newly detected gene family that express in a variety of human cancers.Previous studies have shown that POTEE(POTE ankyrin domain family,member E)was only weakly expressed in normal tissues of prostate and breast,but its expression was significantly elevated in their tumor counterparts.Its biological functions and tumorigenesis mechanisms remains largely unknown.In colorectal cancer,the dysregulation of POTEE are still undefined to our knowledge.Methods:1.Western blot,qRT-PCR and IHC were used to verify the expression of POTEE in colorectal cancer clinical samples and the follow-up data were used to explore the relationship between POTEE and the overall survival of tumor patients.2.We firstly constructed POTEE knockdown and over-expressing stably transfected cell lines,then used MTT,cloning plates,scratch,invasion and Migration)experiments,subcutaneous tumor formation models in nude mice,flow cytometry to detect cell cycle and apoptosis,to clarify the relationship between POTEE and cell proliferation and invasion,cell cycle and apoptosis in colorectal cancer.3.We conducted a genome-wide analysis to globally characterize POTEE-regulated transcriptome changes.4.We used forward and reverse experiments to explore the key genes regulated by POTEE and the activated downstream signaling pathways.Results:1.POTEE is upregulated and predicts poor clinical outcome in CRC patients;2.POTEE plays oncogenic roles in CRC cells;3.RNA-Seq reveals POTEE-regulated signaling pathways in CRC cell;4,POTEE facilitates proliferation through upregulating SPHK1;5.p65 plays as a functional downstream molecular of POTEE/SPHK1 axis;6.Knockdown POTEE could decrease tumor growth in vivo.Conclusions:Our present study sheds light on the oncogenic functions of POTEE in CRC progression.Mechanistically,our results also showed that POTEE might play a critical role in regulating the expression of SPHK1 and followed activation of NF-?B singling,which might act as a novel biomarker and a potential intervention of CRC patients...