The Expression Of Galanin And Its Receptor In Human Pheochromocytoma And The Relationship With The Secretion Of Tumor

PartⅠ:The Clinical Features and The expression of Galanin and its receptors in human pheochromocytomas[Objective]To review and evaluate the distribution,diagnosis and treatment of 156 cases of pheochromocytomas.To research the feature of different pheochromocytoma,and the galanin in serum.To investigate the expression of galanin and its receptors in the tissue of pheochromocytoma.[Methods]Analysis the differences of pheochromocytoma with different function.Detect the galanin in serum with Elisa.Analysis the expression of galanin and its receptors in the tissue of pheochromocytoma with immunohistochemical method.[Results]The hiding function pheochromocytoma without difference on age,sex,innocence/ malignant,adrenal/outside adrenal with the functionality.And has difference in catecholamine of urine in 24hs.the galanin in serum of the hiding function pheochromocytoma is(1.92±0.86)ng/ml[normal value:＜3.5ng/ml],the group of functionality is(1.87±0.84)ng/ml,without any statistics difference with the group of healthy and primary hypertension.6 cases(23.1%) is positive in the expression of galanin in the tissue of hiding function pheochromocytoma with immunohistochemical method,lower than the functionality,123 cases is positive(94.6%).25 cases(96.2%) is positive in the expression of GALR1 in the tissue of hiding function pheochromocytoma,without difference with the functionality,124 cases(95.4%).GALR2 is positive in 9 cases (34.6%),lower than the group of functionality,113 cases(86.9%)。[Conclusions]The function of secrete the catecholamine in pheochromocytoma incorrelate with the level of galanin in serum,it is possible correlate with the the expression of galanin and GALR2 in the tissue.PartⅡ:the primary culture and the identification of human pheochromocytoma cell[Objective]To establish the primary cultural methods of human pheochromocytoma cell.[Methods]Adopt the successive fractionation of the collagenase enzyme digestion to separate and culture human pheochromocytoma cell.Detec the level of catecholamine in cell culture medium,Chromogranin A(CgA) and neuronspecific enolase(NSE) with immunohistochemical method to understand the character of the cell and identify the function,to detect the growth curve of primary culture human pheochromocytoma cell with MTT method.[Results]To establish the primary cultural methods of human pheochromocytoma cell.human pheochromocytoma cell grew comparatively fast from 3^rd to 7^th,cell differentiate in 3^rd to 5^th of culture.we detect the level of catecholamine in cell culture medium,Chromogranin A(CgA) and neuronspecific enolase(NSE) of immunohistochemical is positive,it is proved that the cell can synthetize and secret catecholamine,the cell has different diameter,which can be divide in two kinds of cell,the big one and the small one.the big one has weak expression of CgA,galanin,there is no obviously different expression of NSE,GALR1 and GALR2。[Conclusion]The primary cultural methods of human pheochromocytoma cell can grew and has the function of pheochromocytoma,it can be the basal material of vitro empirical study.The hiding function pheochromocytoma of clinical has little difference with the functionality in culture,it can be any other factors to influence the secretion in vivo. PartⅢ:the influence of Galanin,glucocorticoid to the secretion of catecholamine in the primary cultural human pheochromocytoma cell[Objective]To investigate the influence of galanin and other stimulant to the human pheocbromocytoma cell,to investigate the effection and mechanism of action of galanin and other stimulant to the human pheochromocytoma cell.[Methods]Using the primary cultural methods of human pheochromocytoma cell in partⅡ,different level of Dexamethasone,galanin,TGF,U0126 (inhibiter of ERK kinase),GF109203X(inhibiter of PKC kinase) affect the culture cell,then detect the catecholamine of the medium.Galanin,GALR1 and GALR2 with immunohistochemical method to understand the character of the cell,to detect the growth curve of culture cell with MTT method.[Results]Dexamethasone can increase the secretion of catecholamine in primary cultural human pheochromocytoma cell,and the increasing is dose related,norepinephrine increase 2-3 times,epinephrin increase 2-3 times,make no difference of the proliferation,galanin can increase the secretion of catecholamine in primary cultural human pheochromocytoma cell too,norepinephrine increase 1-2 times,epinephrin increase 1-2 times,but the effect is weak than Dexamethasone.it can enhance the proliferation,this effect can be blocked by U0126 or GF109203X.TGF can enhance the proliferation but depress the secretion of catecholamine.[Conclusion]Human pheochromocytoma cell can secrete catecholamine and this function is correlated with Dexamethasone and galanin.Dexamethasone and galanin can stimulate synthesize and secretion of catecholamine in primary cultural human pheochromocytoma cell,galanin may through ERK or PKC kinase pathway to participate in cell growth and the secretion of catecholamine.GALR2 could be the chief pathway of mediate the physiological functions of galanin,the deficiency of Galanin and GALR2 may be the primary cause of the hiding function pheochromocytoma.