The Study Of ZSCAN10 On Glioma Cell Proliferation Via Activating OCT4 Expression And Regulating Wnt/?-catenin Signaling Pathway

Glioma is the most common malignant tumor in central nervous system.Because of its special location,patients with glioma have poor prognosis and low overall survival.The existing treatment methods for gliomas are limited and the treatment effect is not well,so new treatment for gliomas are urgently needed.Our research group has accumulated some experience in the research of targeted therapy for glioma in the past 10 years.In the big data analysis,we obtained several oncogenes that may be related to glioma,such as USP39,c-Src,TGF-?,P21WAF1,CIP1,PME-PA1 a,etc,and performed K-M on these genes Among them,our research group found that the zscan10 gene expression in K-M analysis of patients with poor prognosis,and the difference was statistically significant(P < 0.01),and less used in tumor research.ZSCAN10 gene,known as zdf206,is located in the 16p13.3 region of human chromosome 16.It encodes a transcription factor composed of 14 zinc finger structures.The amino terminal contains a classical SCAN domain,which plays a regulatory role in dimerization or binding with other zinc finger structural proteins.ZSCAN10 is a transcription factor of zinc finger structure,which maintains the pluripotency of ESCs together with OCT4 and Sox2,but its biological function in glioma is still unknown.It is reported that ZSCAN10 transcription factor,OCT4 and Sox2 form a complex to maintain the pluripotency of embryonic stem cells,and the pluripotency of embryonic stem cells plays a decisive role in organ formation and individual development.It found that the protein molecule encoded by ZSCAN10 gene regulated the expression of OCT4 gene by combining with the promoter region of OCT4 gene,and then affected the pluripotency of embryonic stem cells.The latest research found that the expression of ZSCAN10 gene in the pluripotent stem cells induced by old blood donors was significantly lower than that of the pluripotent stem cells induced by young blood donors,and the genomic stability was poor,and the DNA damage repair program initiated by ROS was abnormal.When ZSCAN10 gene expression was compensated,the stability of cell genome was significantly improved and cell survival was prolonged.However,poor genomic stability,including gene mutation,deletion and duplication,is a common phenomenon in tumor cells.Therefore,based on the above research and analysis,ZSCAN10 gene is very important for the normal development of the body,and its abnormal expression may lead to a variety of development related diseases and may lead to cell carcinomatous transformation.At present,there is a relationship between ZSCAN10 and glioma,but its specific function and molecular mechanism are not clear.In this study,we found that the expression of ZSCAN10 gene in glioma was significantly higher than that in the normal tissue.The results showed that the expression of ZSCAN10 gene in 16 cases of gliomas was significantly higher than that in the control group,and the expression of ZSCAN10 gene in the high grade gliomas was significantly higher than that in the low grade gliomas.Meanwhile,ZSCAN10 gene was highly expressed in many glioma celllines.Immunohistochemical analysis also indicated that ZSCAN10 gene was highly expressed in glioma.These results suggest that zscan10 gene may have important clinical significance in the pathogenesis and progression of glioma.In order to study the function of ZSCAN10 gene in glioma,the RNAi technology was used to knock down the expression of ZSCAN10 gene in glioma cell lines(U251,T98 G and U87MG),the cell proliferation and clonogenesis ability of the three glioma cell lines were significantly reduced,and overexpression of ZSCAN10 in U251 Cell Line could promote the proliferation and clonogenesis of glioma cells.However,the apoptosis experiment showed that ZSCAN10 did not affect the apoptosis of U251 cells.These data suggest that zscan10 gene plays an important role in the growth and proliferation of glioma cells.However,ZSCAN10 had no significant effect on the apoptosis of glioma cells.In order to reveal the molecular mechanism of ZSCAN10 to promote the growth and proliferation of glioma cells,we found that ZSCAN10 gene regulate the expression of OCT4 gene by combining with the promoter region of OCT4 gene,thus affecting the pluripotency of embryonic stem cells.At the same time,a large number of studies have reported that OCT4 plays the role of oncogene in the process of various tumorigenesis and progression,such as liver Cancer,lymphoma,lung cancer,etc.In gliomas,epigenetic analysis showed that the methylation level of the promoter region of OCT4 gene decreased and the expression increased,which promoted the growth and clonal formation of cancer cells.In gliomas,epigenetic analysis showed that the methylation level of the promoter region of OCT4 gene decreased and the expression increased,which promoted the growth and clonal formation of cancer cells.Therefore,we speculate that ZSCAN10 can promote the growth and proliferation of glioma cells by regulating the expression of OCT4 gene.We used RT-q PCR and Western blot to verify that down regulating ZSCAN10 can down regulate the expression of OCT4.Furthermore,we find that overexpressed ZSCAN10 can increase the growth and proliferation of gliomas.We used chromatin immuneprecipitation and immunoluciferase reporter gene experiments to prove that ZSCAN10 can be combined with specific promoter region of OCT4 gene in U251 cell line.This experiment proved that zscan10 can promote the expression of OCT4 gene by directly combining with specific promoter region of OCT4.ZSCAN10 gene promotes the growth and proliferation of glioma cells by promoting the expression of OCT4.In addition,we used RT-q PCR and Western blot analysis to verify that the down-regulation of ZSCAN10 can reduce the expression of Wnt/?-catenin signaling pathway related proteins(?-Catenin,Cyclin D1,TCF)and the expression of OCTA4 GENE at the RNA and protein levels.Therefore,we show that ZSCAN10 can promote the growth and proliferation of glioma cells by activating the expression of OCT4 gene and regulating Wnt/?-catenin signaling pathway.In conclusion,ZSCAN10 can promote the growth and proliferation of glioma cells by activating OCT4 gene expression and regulating Wnt/?-catenin signaling pathway.This study will provide a new insight for uncovering the pathogenesis and progress of gliomas,and may be a new candidate target molecule for the development of glioma targeted drugs.