Correlation Research On Targeted Inhibition Of GOLPH3 And The Sensitivity Changes Of Glioma Cells To Doxorubicin Chemotherapy

Background Brain glioma is the most common malignant tumor in the Department of Neurosurgery,it's poor to treat,and easy to relapse,With a high mortality rate,especially in patients with grade IV WHO glioblastoma(GBM)is the case.Glioma has the characteristics of rapid proliferation,infiltration growth,and Chemoradiotherapy tolerance.Methods for the treatment of gliomas are as follows,traditional craniotomy,directed radiation therapy,chemotherapy,drug therapy,immunotherapy and molecular targeted therapy,etc..In recent years,with the development of molecular biology,the molecular mechanism of tumor cell development has become more and more thorough.The biological behavior of the tumor was found to be related to the overexpression of some oncoprotein,and related signaling pathways were activated.These studies have made it possible for humans to cure the malignant tumor fundamentally.Molecular targeted therapy is aimed at a key molecule in the tumor cells,blocking,inhibiting its function,so as to inhibit or kill tumor cells.Therefore,looking for therapeutic targets is a key point of molecular targeted therapy. GOLPH3 is a newly discovered oncoprotein,It is overexpressed in a variety of malignant tumors,and its expression is positively correlated with the pathological grade of the tumor.GOLPH3 is closely related to the biological behavior of tumor proliferation,division,invasion,migration,chemotherapy resistance,etc.Preliminary results showed that GOLPH3 might be related to the sensitivity of glioma cells to adriamycin and the repair of DNA damage induced by adriamycin.The function of GOLPH3 may be relatedto AKT1 and UBE2 S.GOLPH3 may be a potential target for molecular targeted therapy of glioma.Based on this basis,the related experiments are as follows.Objective To study the changes of glioma cells' sensitivity to doxorubicin after targeted inhibition expression of GOLPH3;Targeted inhibition expression of GOLPH3,then combined with Doxorubicin,study the changes of glioma cells' apoptotic rate;Analysis relationships between AKT1,GOLPH3,UBE2 S.Methods1.Useing siRNA interfering technology,inhibiting GOLPH3 expression of U251,U87 cells line,confirm the transfection efficiency by Western blot and RT-PCR experimental techniques.2.Depending on whether inhibiting the expression of GOLPH3,the U251,U87 cell line were divided into control and treatment groups.Each group were combined with doxorubicin treatment.Through CCK-8 method detecting the changes of doxorubicin IC50values;Cell apoptosis was detected by flow cytometry;Confirm the relationship between AKT1,UBE2 S and GOLPH3 useing Western blot technique.Results1?Confirmed by Western blot,we effectively down regulated the GOLPH3 expression level of U251 and U87 cell lines,P <0.05.2?The doxorubicin IC50 values of U251,U87 cells were(26.53 ± 3.6)?g / ml and(30.62 ± 4.1)?g / ml,after the targeted inhibition expression of GOLPH3,those values become to(7.45 ± 1.4)?g / ml and(12.51 ± 2.5)?g / ml,the difference between those group was statistically significant,P <0.05.3?In U251 and U87 glioma cell lines,targeted inhibition expression of GOLPH3 then combined with doxorubicin treatment,test the apoptosis changes in each group(from the control group,GOLPH3-siRNA-treated group,oxorubicin treatment group to the combined treatment group).Late apoptosis and necrosis both in U251 and U87 glioma cell lines gradually increased,the difference in each group have statistically significant(P <0.05),the combined treatment group compared to the other three groups hasparticularly significant(P <0.01);about the early apoptosis,in U251 cell line,there is a statistically significant difference among each groups(P <0.05),in U87 cell line,there is no statistically significant difference among each groups(P> 0.05).4?Though Western blot experiments,down-regulated the expression level of GOLPH3 in U251 cell line then united doxorubicin treatment,compared to the control group,GOLPH3-siRNA-treated group,AKT1,UBE2 S expression level decreased;doxorubicin treated group,AKT1,UBE2 S expression level decreased;combined treatment group,AKT1,UBE2 S expression level decreased significantly than the control group and the single factor treatment group,all the above results have statistical significant(P<0.05).Conclusions Targeting inhibition Golph3 can increase the glioma cells' sensitivity to Doxorubicin;Targeting inhibition Golph3 then combination with Doxorubicin can obviously induce the glioma cell apoptosis;GOLPH3-AKT1-UBE2 S pathway plays an important role in doxorubicin resistance of Glioma.