Studies On The Expression Of CHD1L And Its Function In Glioma

Objective To verify the high expression of CHD1 L in glioma tissues and cell lines,and analyze its relationship with clinicopathological features and its prognostic value for glioma patients’ survival.To investigated the possible role of CHD1 L in cell cycle,apoptosis,migration and invasion of glioma.Methods 1.Western blot(WB)was used to detect expression of CHD1 L in seven fresh glioma tissues from Grade Ⅱ to Grade Ⅳ.The expression of CHD1 L and Ki-67 was analyzed by immunohistochemistry(IHC)on 81 paraffin-embedded slices.Pearson χ2 test was performed to analyze the associations of CHD1 L expression with clinicopathologic and expression of Ki-67.COX regression analysis and Kaplan–Meier analysis were used to evaluate the contribution of CHD1 L expression on the outcome of glioma patients.2.U87 MG and U251 MG cells were cultured in serum-deprived condition for a period of time,then flow cytometry analysis was used to analyze the cell cycle progression and Western blot was performed to detect protein level of CHD1 L during this process.U87 MG cells were transfected with siRNA targeted on CHD1 L,cell vitality was determined by Cell Counting Kit(CCK8)assay,and cell cycle distribution and apoptosis rate were analyzed by flow cytometry.Moreover,migration and invasion ability was detected by wound healing and trans-well assay.Results 1.Western blot showed that CHD1 L was overexpressed in glioma tissues,and its expression was significantly higher in high-grade glioma tissues than in low-grade glioma tissues.2.IHC data revealed that CHD1 L localized in the nucleus of glioma cells,and its expression increased as the malignant degree increased,which was consistent with Ki-67 expression.Furthermore,there was a positive correlation between CHD1 L and Ki-67 expression in the examined glioma tissues.CHD1 L expression was apparently associated with clinicopathologic grades(P=0.002)and Ki67 expression(P=0.001).Little obvious correlation was observed between CHD1 L expression and patient’s gender,age,tumor location,extent of resection or tumor size in 81 glioma cases.When a multivariate Cox proportional hazard model was constructed,we found that WHO grade,the high Ki67 expression,and CHD1 L expression were prognostic factors of overall survival(P=0.010,0.001,0.041).Kaplan–Meier survival curves indicated that upregulation of CHD1 L was significantly associated with poor overall survival.Combined these results together,we inferred that high expression of CHD1 L could be a strong determinant of poor prognosis in glioma.3.After serum starvation,U87 MG and U251 MG cells were arrested in the G0/G1 phase,then upon serum refeeding,cells in the S phase and expression of CHD1 L increased,expression pattern of CHD1 L was consistent with that of PCNA and cyclinD1.4.Knockdown of the expression of CHD1 L inhibited the proliferation of glioma cells.Loss of CHD1 L expression induced apoptosis in glioma cells.CHD1 L perhaps participated in EMT and affected certain actions of glioma cells,which was related to the migration and invasion of glioma.Conclusions 1.Western blot and immunohistochemistry analysis showed that CHD1 L was overexpressed in glioma tissues and glioma cell lines.In addition,the expression level of CHD1 L was positively correlated with glioma pathologicalgrade and Ki-67 expression.Kaplan–Meier curve indicated that high expression of CHD1 L may result in poor prognosis of glioma patients.2.Supression of CHD1 L in glioma cells was shown to induce cell cycle arrest and increase apoptosis.In addition,knockdown of CHD1 L significantly accelerated migration and invasion ability of glioma cells.Together our findings suggest that CHD1 L is involved in the progression of glioma and may be a novel target for further therapy.