| background:Transmembrane protein(TMEM)is an integral membrane protein that spans biological membranes.It is known that members of the transmembrane protein family are related to the progression of many tumors.However,there is no research on the mechanism of TMEM44 gene in renal clear cell carcinoma(KIRC).Therefore,this study aims to analyze the effect of TMEM44 gene on KIRC cell proliferation,apoptosis and migration,and to explore its underlying molecular mechanism.method:1.We first used bioinformatics to analyze the expression difference of TMEM44 gene in KIRC and the survival prognosis of patients;used single and multivariate COX regression to analyze the correlation between TMEM44 gene and patient prognosis,and analyzed the prognostic value of TMEM44 in KIRC;through ss GSEA Algorithm predicts the molecular mechanism of the influence of TMEM44 gene on the biological activity of KIRC cells.2.The expression difference of TMEM44 gene in KIRC tissue was confirmed by PCR experiment,immunohistochemical method and western blot experiment;the expression difference of TMEM44 gene in KIRC cell line was confirmed again by PCR experiment and western blot experiment;then knocking down TMEM44 expression was passed through CCK8 Observe the changes in the proliferation ability of 786-O cells by experiments and Ed U experiments,observe the changes in the migration ability of 786-O cells by wound healing experiments and cell migration experiments,and observe the changes of TMEM44 on the cell cycle and apoptosis of786-O cells by flow cytometry;finally Using stably transfected 786-O cells knocking down the expression of TMEM44 to construct a subcutaneous tumor survival model in nude mice to analyze the effect of TMEM44 on the growth ability of 786-O cells in vivo xenograft tumors.3.After knocking down the expression of TMEM44,explore the changes of PI3K/AKT/m TOR signaling pathway-related proteins and the changes of proliferation and apoptosis-related proteins in 786-O cells.result:1.Bioinformatics analysis of the high expression of TMEM44 gene in KIRC is related to the poor prognosis of patients;the ss GSEA algorithm is used to predict the close relationship between TMEM44 gene and PI3K/AKT/m TOR signaling pathway in KIRC;PCR experiments,immunohistochemical methods and western blot Experiments also confirmed that TMEM44 gene was significantly highly expressed in KIRC tissues and cell lines(P<0.05).2.After knocking down the expression of TMEM44,CCK8 experiment and Ed U experiment confirmed that the proliferation ability of 786-O cells could be significantly inhibited;wound healing test and transwell migration experiment confirmed that the migration ability of 786-O cells may be obviously inhibited;flow cytometry confirmed that 786-O cell cycle was arrested in the G1 phase,and the apoptosis rate was obviously increased;the subcutaneous tumor preservation experiment in nude mice also proved that the tumor growth ability was significantly inhibited after knocking down the expression of TMEM44(P <0.001).3.After knocking down the expression of TMEM44,the protein expression levels of p PI3 K,p AKT,and pm TOR in 786-O cells in the normal group were obviously reduced,the expression levels of proliferation proteins MMP2,MMP9,and MMP13 were obviously reduced,the expression levels of BCL2 protein were obviously reduced,and the expression levels of Caspase3 and BAX were obviously increased.(P <0.001).conclusion:1.Bioinformatics analysis and clinical sample experiments confirmed that TMEM44 gene is highly expressed in KIRC and is associated with poor prognosis of patients,which may be an independent prognostic risk factor for KIRC2.TMEM44 is a tumor-promoting gene in KIRC.It has been confirmed by in vivo and in vitro experiments that it promotes the proliferation of KIRC cells,and in vitro experiments also confirm that it promotes the migration of KIRC cells and inhibits the apoptosis of KIRC cells.3.TMEM44 affects the proliferation,apoptosis and migration of KIRC cells through the PI3K/AKT/m TOR signaling pathway. |
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