Effect Of CARMA3 Gene On Proliferation And Apoptosis Of Glioma Cells

Objective explore the molecular mechanism that CARMA3 gene regulate the proliferation and apoptosis of glioma cell,and discuss the possibility of using the CARMA3 gene as a potential therapeutic target for glioma.Methods(1)acquire the glioma specimens and non-tumor brain tissue(paraneoplastic tissues)by surgery,and detect the expression of CARMA3 gene in tumor tissues and paraneoplastic tissues by RT-q PCR.(2)according to the CARMA3 gene sequence of gene bank,design the si RNA interference sequence of CARMA3 m RNA,filtrate si RNA interference sequence with high interference efficiency.(3)set a blank control group,a negative control group,and a CARMA3 si RNA group separately.Detect cell proliferating changes of different experimental groups by CCK-8 kit,apoptosis situation by flow cytometry.Cell proliferation,apoptosis-related proteins(Bcl-2,Cyclin D1,P53),along with expressions of NF-κB protein and CARMA3 protein are detected by Western blot.(4)By regulating the activity of NF-κB pathway in the blank control group and CARMA3 si RNA group,detect the proliferation and apoptosis of each group by CCK-8 and flow cytometry as well,along with the relevant proliferation and apoptosis protein level by Western blot.Results(1)The result of RT-q PCR showed that the expression of CARMA3 gene in glioma tissues was significantly higher than that in paraneoplastic tissues.(p<0.01)(2)Design and screen out si RNA interference sequences of CARMA3 m RNA with high interference efficiency CARMA3 si RNA3.(3).The expression of P-P65 and CARMA3 protein in U251 glioma cells transfected with CARMA3 si RNA3 interference sequence was weaker than that of the blank control group and the negative control group(p<0.01).The proliferation ability of the cells was significantly weaker than that of the blank control group and the negative control group(p<0.01).Compared with the blank control group and the negative control,the apoptosis ability was significantly enhanced(p<0.01).After silencing CARMA3 gene,the expression of Cyclin D1 and Bcl-2 in U251 glioma cells was significantly decreased,and the expression of P53 was significantly enhanced(p<0.01).(4)The addition of NF-κB promoter LPS increased the proliferation and the expression of Bcl-2 and Cyclin D1(p<0.01),but decreased P53 expression and ability of cell apoptosis in the CARMA3 si RNA group cells(p<0.01),After adding NF-κB antagonist PS-341 to the blank group,the apoptosis and the expression of P53 was enhanced(p<0.05),theexpression of Bcl-2 and Cyclin D1 and cell proliferation ability was weakened(p<0.01).Conclusion Transfecting the U251 glioma cells with CARMA3 si RNA sequence can effectively reduce the expression of CARMA3 m RNA and CARMA3 protein.CARMA3 si RNA can effectively induce apoptosis of U251 glioma cells by regulating the NF-κB pathway.This study provide experimental support for the CARMA3 gene as a potential therapeutic target for glioma and silencing of the CARMA3 gene as an effective means of treating glioma.