The Experimental Study Of STIP1 Promote Glioblastoma Invasion By TRAP1-AKT Signaling Pathwag

Objective:Glioma is the most common malignant tumor with intracranial primary malignancy.It has the characteristics of high morbidity,recurrence rate,mortality and low cure rate.More and more studies tend to molecular targeted therapies,so seeking specific targets,and developing new treatment plans,which may lead to breakthrough in the treatment of glioma.In recent years,many studies have confirmed that the phosphorylated stress induced protein 1(STIP1)is involved in the occurrence and development of a variety of tumors.In this paper,we examined the differences of expression in human glioma tissue samples and normal brain tissues.Study on the effects of STIP1 on the proliferation and invasion of glioma cell lines;To explore the relationship between STIP1 and TRAP1 in glioma.To lay a theoretical foundation for the diagnosis of glioma and molecular targeted therapy.Method:41 glioma tissue which were parts of samples choosed from 2000 to 2016 years collected from the First Affiliated Hospital of Soochow University.That included 35 cases different levels of glioma tissue,6 cases of normal brain tissue.We extract RNA of brain tissue,then using real-time quantitative PCR(qRT-PCR)technology,detect STIP1 gene express in different grade glioma tissues and nomal brain tissue.At the same time the differences in protein levels were detected by Western Blot and immunohistochemistry.The lentivirus(siRNA)was transfected into glioma cell lines U87 and U251,and then MTT was used to detect the proliferation ability of cells.Transwell test detects invasion ability.Western Blot was detected before and after transfection with the expression of 1,TRAP1,AKT,p-akt and MMP2.Result:1.The level of gene and protein in human glioblastoma was significantly higher than that in non-cancerous brain tissue.Using qRT-PCR technolog,we analyzed the STIP1 in 6 non-cancerous brain tissue samples and 35 samples with different levels of human glioma.The results indicated that there was no significant difference between the lower grade glioma and the non-cancerous brain tissue(P>0.05),and there was a significant difference between the non-cancerous brain tissue and glioblastoma(P<0.05).Immunohistochemistry and westernblot assay showed that the expression of STIP1 in glioblastoma was higher than that of noncancerous brain tissue.2.Down-regulation of STIP1 expression inhibited proliferation and invasion of glioma cell lines.By using lentivirus stable transfection technique,after down-regulating STIP1 expression in U87 and U251 glioma cell line,the proliferation of U87 and U251 glioma cell line was significantly inhibited by MTT assay,showed that the invasion ability was decreased by transwell assay.3.After down-regulation of STIP1 expression,the expression of TRAP1p-AKT MMP2 protein decreased.Lentivirus-transfected U87 and U251 cells down-regulated the expression of STIP1.Western blot showed that the expression of TRAP1 protein decreased and the expression of invasion associated protein p-AKT-1 MMP2 decreased.Conclusion :(1)the expression level of STIP1 in human glioblastoma is significantly higher than that in non-cancerous brain tissue.(2)After down-regulation of STIP1,it can inhibit the proliferation and invasion of glioma cells.(3)STIP1 can regulate the glioblastoma invasion of cells through TRAP1-AKT pathway.