Effect Of Corticotropin Releasing Factor (CRF) On Human Glioma Cell Line U87Proliferation

Background: Corticotropin-releasing factor(CRF) is a hypothalamusneuropeptide containing41amino acids, mainly regulate the hypothalamic-pituitary-adrenal axis function when the body under stress, play acoordination of the autonomic nervous system, endocrine, immune system,physiological and behavioral functions. In addition, CRF on pregnancy,preterm birth, skin, digestive system, cardiovascular system, reproductivesystem, etc. has a modulatory role. In human endometrial cancer, ovariancancer, breast cancer, skin cancer, liver cancer and central nervous systemtumors were found CRF over-expression. The animal experimental modelshowed that the CRF family play a certain role in tumor proliferation,growth, and metastasis. The role of the CRF family of neuropeptides in thecentral nervous system tumor research is still relatively limited, especially inglioblastomas, both at home and abroad are not explicitly reported. We haveused immunohistochemistry and RT-PCR method Were confirmed by theCRF receptor CRFR1expression in glioma cells. CRF as a kind of tumorpromoting factors, there must be a potential role in the development ofgliomas. it will be confirmed in further studies.Objective: Gliomas are the most intracranial malignant tumor, becauseof its highly invasive, great harm to human health, the treatment effect is notideal. From the point of view of biological molecules to study the occurrenceand development of the glioma has become a hot spot. The role of CRF as apro-tumor growth factor in the development of gliomas has not been clearlyexplained. The purpose of this study is to explore the added containingdifferent concentrations of CRF,the factor of U87glioma cell proliferation,Thus confirming the CRF play a catalytic role in the growth of glioma cellsU87cells.it will be provide new content for the further study of the occurrence and development of gliomas.Method:Cultured glioma cells U87, establish the experimental groupand control group,using three different methods to detect the proliferation ofglioma cells U87.(1)At different incubation time (24hours,48hours,72hours), usinginverted microscope to Observe the growth conditions of the experimentaland control groups, to judgment whether CRF play a catalytic role in theprocess of tumor growth.(2)In the experimental group, adding different concentrations of CRF inthe cells culture medium, the concentration of CRF were10-9,10-8,10-7mol/L, The use of MTT method to detect the absorbance values of A in differentaction time (24hours,48hours,72hours), determine the different CRFconcentration and time on the proliferation of glioma cells U87.SPSS13.0statistical software to analyze the data.(3)Enzyme histochemical method stained the mitochondria, through thechanges of mitochondrial staining to reflect the role of CRF in tumor cellmetabolism.Results:(1)Inverted microscope observations show: the experimental groupcompared with the control group,Join10-7mol/L CRF were cultured for24hours,48hours,72hours, U87cell in the experimental group is better, thenumber of adherent cell with a significant increase. the morphology of cellswith diversity, good adhesion between cells, cells tightly covered in theculture bottom.(2)In the U87cell proliferation assay, we found that10-9,10-8,10-7mol/L CRF on the growth of glioma cells U87can promote the proliferativeeffect. Using ANOVA for repeated measurement design data, We derive thesphericity test results, P=0.227>0.05. satisfy the covariance matrix of thespherically symmetric conditions, no correction on the results. At the sameconcentration, the effect of different time, between in24hours,48hours,72hours,Cell proliferation is difference (F=65, P <0.05), there are Statistical significance.At the same time, different concentrations of CRF comparedwith control groupAnd between the CRF10-9,10-8,10-7mol/L, there arestatistically significant (P<0.01). there are statistically significant.Compared with the control group, joined the concentration of10-9,10-8,10-7mol/L CRF,cell absorbance value A increases with the concentration ofCRF, and showed a dose-effect relationship.(3)Enzyme histochemical staining showed that in certain drug concentration range as the experimental group to join the CRF concentration increases continuously,In24hours,48hours after intracellular mitochondria color was deep-ened,distribution than the control group,nuclei differentiation is more active.Conclusions:CRF can promote the value of U87glioma cells,And rendered within a certain range of time and dose dependent.