Research On The Expression Of Pirh2 And Its Function In Glioma

Objective 1.To study the expression of Pirh2 in glioma patients and to explore the relationship between Pirh2 and glioma development and progression.2.To study the molecular mechanism of Pirh2 on cell apoptosis and proliferation at the cellular level,and to further study the molecular mechanism of glioma progression,and provide a promising therapeutic target in the treatment of glioma.Methods 1.The expression of Pirh2 in normal brain tissue and glioma tissues was detected by Western blot(WB)method at the tissue level.we analyzed expression of Pirh2 and Ki-67 in 90 glioma samples by immunohistochemistry.The relationship between the expression of Pirh2 with the clinicopathological parameters and Ki-67 expression was analyzed by Pearson ?2 test.2.At the cellular level,the expression of Pirh2 in glioma cell lines was detected by WB,and the highest expression cells were selected.We used siRNA oligos to silence Pirh2 expression in U87 MG cells.we performed western blot analysis to examine the expression of apoptosis-related proteins,such as Bcl-2 and cleaved caspase-3.The changes of apoptosis were detected by flow cytometry.3.The expression of PCNA and cyclin D1 was detected by WB in U87 MG cell line.The cell cycle was detected by flow cytometry,CCK-8 and clone formation assay were used in order to detect the proliferation of glioma cells.Results 1.Western blot and immunohistochemistry revealed a significantly increased of Pirh2 expression in glioma tissues when compared with normal brain tissues.and the expression of Pirh2 was gradually increased with the increase of glioma grade.Moreover,we observed a strong correlation between Pirh2 expression and the degree of malignancy(P<0.003).In addition,the expression of Pirh2 was positively correlated with that of proliferation marker protein Ki-67.In glioma cells,correlation analysis showed that Pirh2 expression was positively correlated with Ki-67 expression.Kaplan-Merier analysis showed that the increase of Pirh2 expression in glioma cells was associated with a reduced overall survival rate(P <0.01).2.Among the cell lines,Pirh2 was most abundantly expressed in U87 MG cells.After inhibiting the expression of Pirh2 in U87 MG cells,the levels of cleaved and caspase-3 were increased,and the levels of Bcl-2 were decreased.and flow cytometry analysis showed the apoptosis of the cells was increased.3.In the U87 MG cells,interference of Pirh2 resulted in apparently reduced expression of PCNA and cyclin D1,suggesting that cell division was stagnated.CCK-8 test and clone formation test showed that cell proliferation was decreased.Conclusions 1.The expression of Pirh2 was positively correlated with the WHO grade,and showed a significant positive correlation between Pirh2 and Ki-67.The expression of Pirh2 was correlated with the clinical prognosis of glioma patients,suggesting that Pirh2 was involved in the development of glioma.2.Pirh2 could inhibit the apoptosis of glioma cells and promote the proliferation of glioma cells.Silencing Pirh2 expression could promote apoptosis and impairs the proliferation of glioma cells,so Pirh2 may be a new target for glioma treatment.