Effects Of Embryonic Stem Cell Microenvironment On Neuronal Tumor Cells

The first part research purposes to explore the feasibility and possible mechanism of stem cell microenvironment for the treatment of Parkinson's disease.Methods:The experiment was divided into four groups.SH-SY5Y cells cultured in vitro were used as blank control group(SH group);SH-SY5Y and ES were indirectly co-cultured group(SE group)at 3:1 ratio;SH-SY5Y and ES were indirectly co-cultured at 3:1 plus MPP chemotherapy drugs(SEM group);SH-SY5Y plus MPP chemotherapy positive control group(SM group);All groups were harvested SH-SY5Y cells after 72 hours of culture,respectively,cell proliferation experiments,cell death Experiments,reactive oxygen species detection,quantitative real-time RT-PCR(qPCR),and Western Blot(WB)were performed to investigate the effect of embryonic stem cells on SH-SY5Y cell.The effects of cell biology functions and JNK/P53/Bax signaling pathways were compared and the differences between the groups were compared.Results:ESc microenvironment promoted the apoptosis of SH-SY5Y cells induced by MPP+and accelerated the death of SH-SY5Y cells.The expression of reactive oxygen species(ROS)in ES and SH-SY5Y co-culture groups increased significantly,and the expression of apoptosis-related proteins JNK,P53 and Bax increased.Conclusion:ESc can promote the apoptosis of SH-SY5Y cells through the JNK/P53/Bax pathway,which has the potential to treat neuroblastoma.The second part research purposes:To explore the possible mechanisms and treatmentmethods of reducing the malignant degree of human glioma by the microenvironment of embryonic stem cells.Methods:Choose the nervous system of the most common malignant tumor cell lines,glioma cell line(U118),the experiment is divided into four groups,and a total in cultured glioma cells alone(U118)as the blank control group(GB);U118 directly co-cluture with ES in accordance with the ratio 3:1 trained group(GE);U118 and ES according to 3:1 after co-culture with PI3K agonist group(GA);U118 plus for mo thiazole amine as the positive control group(GT);collecting U118 cells of four groups after culture 72 h,analysis biology function of U118 glioma cell and PI3K/AKT signal pathway and compare differences between groups by cell proliferation experiment,cell apoptosis experiment,reactive oxygen experiment,microvascular experiments,quantitative real-time RT-PCR(qPCR),protein immunoblot experiment(Western Blot,WB).Results:GE co-culture group and chemotherapy drugs in the control group compared with the blank control group in GB,co-culture group and chemotherapy drugs weakened cell proliferation ability,apoptosis increased,reactive oxygen species increased,angiogenesis mimicry formation reduce or even disappear,promoting tumor proliferation proteins as Cyclin D and Cyclin B decreased significantly,and promoted tumor cell apoptosis and inhibit proliferation of related proteins such as the expression of BAX,BCL-2,P53,Caspase3,Gsk-3 B and P21 and P27 significantly increased,with significantly statistical differences.While in PI3K/AKT pathway,expression of positive regulating factor as PI3K,PDK,AKT,mTOR decreased,such as negative control factor expression of PTEN increased,differences were statistically significant;adding PI3K agonist group(GA)compared with GE group,expression of positive adjustment factor increased and negative adjustment factor reduced,the difference was statistically significant.Conclusion:The embryonic stem cell microenvironment reduced the malignant degree of glioma by down-regulating PI3K/AKT signaling pathway,inhibiting tumor cell proliferation,inhibiting protein synthesis,and promoting tumor cell apoptosis.