| BTCC is the most common urinary system carcinoma. Up to date, however, the cellular and molecular mechanisms leading to carcinoma-tous change of bladder cell remain unclear. Increasing evidence exists that carcinogenesis must be understood in terms of accumulation of mutationns in regulatory genes. Including activation of oncogenes and inactivation or loss of tumor suppressor genes. Among the available candidates, the p53 gene has come to be known as a master guardian of the genome. Recently, a new gene with significant homology to p53 has been discovered and named p73 ( Kaghad et al, 1997). This gene mapped to the short arm of chromosome 1. It has also been reported recently that p73 can at least when overproduced activate the transcription of p53 - responsive genes and inhibit cell growth, in a p73 -like manner by inducing apoptosis. But unlike p53, no significant mutation of the p73 has been found in human tumors. This is different from classics and - oncogene. No data describing the expression of p73 in BTCC in correlation to the mutation expression of p73 have so far been reported. We therefore performed immunohistochemistry and TUNEL for the p73 protein and apoptosis expression and analyzed thecorrelation between p73 and clinicopathologic factors. MATERIAL AND METHODSA total of 44 BTCC removed by partical cysteetomy, radical cys-teetomy, and transurethral resection of bladder tumor. To be freezen in liguid nitrogen and preservated in -70℃. No patient received pre-operative c'hemo - or radio - therapy but regularly irrigation of bladder after operation0 Patients included 34 men and 10 women, ages 32 -79 years (mean age 55 years). Histopathological and clinical classification of tumors was performed according to the criteria of WHO and UICC. All patients were clinically followed -up. All case were fixed in 10% phophate -buffered formalin or acetone. For each case, all a-vailable hematoxylin and eosin stained sections wer reviewed. The rabbit polyclonal p73 antibody and In situ cell apoptosis detection kit were obtained from SANTA CRUZ Biotechnology. Immunohistochemi-cal case was performed on acetone for 10 min. The endogenous peroxide activity was blocked with 3% hydrogen peroxide for 5 minutes. After the sections were incubated with 10% normal goat serum to reduce nonspecific binding they were treated with anti - p73 antibody (diluted 1: 50 ) at 37℃ for 60 minutes. Immunostaining was performed by the strepvidin - biotin being incubated at 37℃ for 10 mi-nates. Apoptosis case were performed on 10% neutrality formalin for 25 minute the endogenous peroxide active was blocked with 3% hydrogen peroxide for 10 minutes then put into 0. 2% triton - x - 100 at 37℃ for 15 minute. Apoptosis cases were performed on the mixture include TdT, Diction -11 - dVTP and labeling Buffer at 37℃ for 10 mimutes. Soaked in 2xSSC at 37℃ for 5 minutes, treated by Avidin -HPP, at 37℃ for 60 minutes. This method use a histofine sap - pokitstained was visualized with diaminobenzidine tetrachoride. The nuclei were counter stained lightly with hematoxylin the phosphate - buffer saline was substituted for primary antibody or TdT as the negative control. P73 and Apoptosis expressions was evaluated as positivive if the stained cells were determined by the chi - square test. Compared mean by analysis of variance.RESULTSImmunoreaction to p73 and Apoptosis was localized in the nuclei of neoplastic cells. Overall, 55% (19 of 44) of the surgically resected tumors were positive for p73, for apoptosis nuclear staining was detected in 23 of 44 tumors ( 52% ). Histologically 8 normal mucous membrane of bladder tissues were negative for p73 and apoptosis. In our study, p73 and Apoptosis were positively related in tumor grade (p <0.05). No significane correlation was found between p73. for apoptosis expression and clinical stage ( p > 0. 05 ) . There was good correlation for the expression between p73 proteins and apoptosis in BTCC0 Approximated 71% of p73 positive BTCC. (19 out of 24) also displayed positive reactio...
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