Correlation Of INPP4B Gene To Proliferation,Metastasis And Apoptosis Of Ovarian Cancer Cells

Background: Ovarian cancer is the highest fatal gynecological cancer in the world.Due to the early clinical symptoms are not significant,most of the ovarian cancer patients have been diagnosed late clinical stage,while advanced ovarian cancer patients with very low survival rate.Early diagnosis of ovarian cancer less,there is no perfect test method.Late treatment includes the main means of treatment of chemotherapy,radiotherapy and other methods,but the lack of related targeted therapy.Because of the high specificity of detection methods can not effectively detect the disease in the crowd,it is difficult to detect ovarian cancer in the crowd.Inositol polyphosphate4-phosphatase type(INPP4B)is a potential tumor suppressor and 3-phosphoinositide kinase(PI3K)is a key kinase of myo-inositol and phosphatidylinositol and binds to AKT(Protein kinase B,PKB)constitute the PI3 K / AKT signaling pathway,play an important role in cell proliferation and growth.Recent studies have reported that INPP4 B gene involved in the PI3 K / AKT pathway in breast cancer and melanoma expression.Objective: To study the expression of INPP4 B gene in ovarian cancer.In order to find a new target for ovarian cancer,we provide more protocols for clinical treatment of ovarian cancer.Methods: 1,the information of 356 cases of INPP4 B gene in human gene pool was screened by bioinformatics TCGA library,and the ovarian tissues of normal ovarian tissue and ovarian cancer patients were analyzed to study the regulation of INPP4B gene in cell proliferation and metastasis effect.(Real-time Quantitative PCR)q PCR analysis of 40 cases of clinical cases of ovarian cancer and normal ovarian tissue INPP4 B m RNA expression levels.2,According to the nucleotide sequence of the target gene INPP4 B and its negative control interference sequences,specific sh RNA plasmids were designed to knock INPP4 B and over-express INPP4 B with GV230 plasmid.In vitro experiments by INPC4 B gene transfection interference technique using lipofectamin2000 liposome INPC4 B gene downregulation and INPP4 B gene transfection into A2780 and SKOV-3 cells,the use of Western Blot experimental technique to detect INPP4 B gene in ovarian cancer Cell expression.3,Clone formation assay and MTT method to test the proliferation of ovarian cancer cells A2780 and SKOV-3 under different expression of INPP4 B gene.4,Using flow cytometry and DAPI staining to detect the apoptosis of ovarian cancer cells under different expression of INPP4 B gene.The relevant apoptosis protein was used to verify the expression of its apoptosis.5,Through the cell scratch test to verify the different expression of INPP4 B ovarian cancer cell migration and migration.The relevant transfer protein is used to verify the expression of its metastasis.6,The proliferation and proliferation of ovarian cancer cells overexpressing INPP4 B gene after cisplatin treatment were detected by MTT and cell clone formation assay.7,Western Blot method was used to detect the expression of related genes in PI3 K / AKT signaling pathway.Results: 1,TCGA database data can be found,the use of Junc BASE parameter settings,screening out the normal ovarian tissue and ovarian cancer INPP4 B gene content in the m RNA expression of water compared.The experimental results show that the content of INPP4 B in normal ovarian tissue is higher than that of ovarian cancer.Also found that as the disease aggravated G1-G3 phase,INPP4 B gradually decreased.As shown in the results of the 5-year survival rate,the 5-year survival rate of INPP4 B gene-overexpressing patients is higher than that of patients with low INPP4 B gene expression.(Real-time Quantitative)PCR results found that the level of INPC4 B in ovarian cancer ovarian tissue at the m RNA level is lower than the content of INPP4 B in normal ovarian tissue.2,The transfected INPP4 B gene interference technique successfully obtained over-expression of INPP4 B and low expression of INPP4 B stable transfected A2780 and SKOV-3 cell lines.Western Blot results showed that the target gene was successfully introduced into the relevant cell lines and expressed accurately.3,The results of colony formation assay and MTT assay showed that INPP4 B gene was overexpressed and inhibited the proliferation of tumor cells.However,knockdown of INPP4 B gene promoted the growth of tumor cells.4,The results of flow cytometry and DAPI staining showed that INPP4 B overexpression promoted the apoptosis of ovarian cancer cells,while the number of apoptotic cells in knockdown group decreased compared with the control group.Western Blot results showed that over-expression of INPP4 B gene and the increase of Caspase9,a pro-apoptotic protein,decreased the content of anti-apoptotic protein BCL-2.The low level of INPP4 B gene expression of the pro-apoptotic protein Caspase9 relative negative control group decreased anti-apoptotic protein BCL-2 content increased.5,Scratch experiment results showed that over-expression of INPP4 B gene inhibited cell migration,whereas knockdown group increased the metastasis of tumor cells relative to the control group.Western Blot results showed that INPP4 B gene overexpression,MMP2 and MMP9 protein content decreased,INPP4 B low expression group increased.6,The results of MTT and cell clone formation showed that the proliferation of ovarian cancer cells overexpressing INPP4 B gene after cisplatin treatment was significantly inhibited compared with the control group.7,Western Blot experimental results confirmed that INPP4 B gene expression is involved in the regulation of PI3 K / AKT signaling pathway.Conclusion: The proliferation and metastasis of ovarian cancer cells were significantly inhibited by over-expression of INPP4 B gene and the apoptosis of ovarian cancer cells was increased.While knockdown of INPP4 B expression enhanced its proliferation and metastasis relative to the negative control.This indicates that the gene plays an important role in the proliferation and growth,migration and apoptosis of ovarian cancer cells.INPP4 B gene as a potential tumor suppressor gene,or in the treatment of ovarian cancer has a new way of thinking and methods.