PURPOSE: Recently, it is suggested that the inhibitian of apoptosis is associated with tumorigenesis of colon. Bcl-2 gene is an important inhibitory regulator of apoptosis, and bcl-2 acts antagonistly with the wild type p53 gene, one of the tumor suppressor genes, in apoptosis. To detnmine the role of bcl-2 and p53 gene in colonic tumorigenesis, we performed the study. MATERIALS AND METHODS: We tested the tissue obtained by polypectomy and surgical resection by immunohistochemical staining for Bcl-2 and p53. RESULTS: We found that in normal colonic tissue, the Bcl-2 was sparcely expressed, and the p53 was expressed sporadically. The rate of positivity of staining was below 5%. However, in colonic adenoma and colon cancer tissue, Bcl-2 and p53 were expressed more than in nonnal colonic tissue(p<0.05). (Scoring in Colonic adenoma: Bcl-2 6.2+/-1.1, p53 5.7+/-1.0; Scoring in Colonic carcinoma: Bcl-2 4.7+/-1.0, p53 8.3+/-0.9) CONCLUSION: Our results suggested that the bcl-2 and p53 play an important role in colonic tumorigenesis.
Adenoma* ; Apoptosis ; Carcinogenesis ; Colon* ; Colonic Neoplasms ; Genes, bcl-2 ; Genes, p53* ; Genes, Tumor Suppressor

PURPOSE: This study was undertaken for evaluation of the extent of apoptosis and expression of the bcl-2 gene and p53 gene as prognostic factors in breast cancer. MATERIALS AND METHODS: The extent of apoptosis was determined by the 3' end-labeling method of DNA in tissue sections and the expressions of bcl-2 and p53 was determined by immunohistochemical staining in 41 cases of breast carcinomas. RESULT: bcl-2 was correlated with ER positivity (p=0.005), with nuclear grade (p=0.001), and with tumor size (p=0.019), whereas it was inversely correlated with apoptosis (p= 0.018). No association was found with axillary nodal status. Apoptosis was not associated with clinicopathological parameters, such as ER positivity, nuclear grade, tumor size, and lymph node status. p53 was not associated with above clinicopathological parameters. No relationship between the expression of bcl-2 and p53 was found. CONCLUSION: These results suggest that bcl-2 protein inhibit apoptosis, and its expression is associated with favorable clinicopathological features.
Apoptosis* ; Breast Neoplasms* ; Breast* ; DNA ; Genes, bcl-2 ; Genes, p53 ; Lymph Nodes

OBJECTIVETo observe the cell apoptosis after tractive spinal cord injury in rats, determine expression of apoptosis correlative genes, and study the molecular mechanism of cell apoptosis.

METHODSThe T(13)-L(2) spinal cord of rats was injured by traction after the amplitude of P1-N1 wave decreased to 70% in postoperation than in preoperation through cortical somatosensory evoked potential (CSEP) monitor. Then rats were killed in 30 min, 6 h, 1, 4, 7, 14 and 21 d respectively after operation (n = 4). Cell apoptosis was examined by the flow cytometer and terminal deoxynucleotidyl transferase-mediated DUTP-biotin nick end labeling (TUNEL) reaction, the expression of p53, bax and bc1-2 genes was tested with immunohistochemistry.

RESULTSThe flow cytometer test and TUNEL method showed that the apoptosis cell ratio raised in 6 h and reached at peak in 7 d after injury, and then declined till 21 d, they showed significant difference (P < 0.05, 0.01). TUNEL method showed that injured group had a large number of apoptosis glial cells in white matter. Immunohistochemical staining showed that the positive expression of p53, bax and bc1-2 protein raised at 6 h, expression of p53 protein reached at peak in 4 d, bax and bc1-2 protein reached at peak in 7 d after injury. Compared with control group and laminectomy group, the injured group showed significant difference (P < 0.05, 0.01).

CONCLUSIONThere is cell apoptosis phenomenon after tractive spinal cord injury in rats. Morphology indicates that apoptosis includes neurons and glialcytes, which is an important form of cell death and pathological changes in secondary lesion period after tractive spinal cord. There exist high expression of apoptosis correlative gene p53 and bax after spinal cord injury, they may play an important role in reduction of cells to apoptosis.

Animals ; Apoptosis ; genetics ; Disease Models, Animal ; Female ; Gene Expression ; Genes, bcl-2 ; genetics ; Genes, p53 ; genetics ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; genetics ; pathology ; bcl-2-Associated X Protein

BACKGROUND: Scid.adh is a recently developed murine thymic lymphoma cell line, which has been used as in vitro model for the study of double negative stage III thymocytes. In this study, we compared the expression profile of a number of genes and proteins, which are tightly related to T cell development and apoptosis, in thymic lymphoma cell lines, R1.1, EL-4, and Scid.adh for the developmental staging. METHODS: We examined the expression of development marker genes and proteins in three lymphoma cell lines by flow cytometry and RT-PCR. In addition, the expression of apoptosis-related molecules including bcl-2, bax and Fas was also investigated. RESULTS: As previously reported, Scid.adh cell line expressed CD8 and CD25 but not TCR alpha chain, while R1.1 cells expressed TCR alpha chain and both CD4 and CD8 transcripts. These suggest that R1.1 might be in double positive stage, and low level of CD44 expression and the absence of CD25 support this suggestion. In contrast, EL-4 cells showed high level of TCR alpha chain transcript, and low-level of CD4 expression, suggesting that EL-4 is in more mature stage than R1.1. Further, this suggestion was supported by the lack of mT-20 in EL-4 cells, which is expressed in the immature thymocytes, and Scid.adh and R1.1 cell lines, but not in the terminally differentiated thymocytes and peripheral T cells. Among the apoptosis-related gene, transcripts of bcl-2 gene were detected in both R1.1 and EL-4 but not in Scid.adh cells, while bax was expressed in all cell lines. Fas expression was the highest in EL-4 cells and low in Scid.adh cell line. CONCLUSION: R1.1 cell may represent double positive stage, and EL-4 is more differentiated cell line. In addition, Scid.adh and EL-4 cell lines are suspected to be useful for the study of function of bcl-2 family and Fas during the thymocyte development, respectively.
Apoptosis ; Cell Line* ; Flow Cytometry ; Genes, bcl-2 ; Humans ; Lymphoma* ; T-Lymphocytes ; Thymocytes

OBJECTIVE: The study was designed to investigate the expression of Bcl-2 and CD-44 molecules in cervical cancer and compare the results with known clinical prognostic factors, and lastly to define the roles of Bcl-2 and CD-44 molecules in tumorigenesis and metastasis of cervical cancer. METHODS: A total of 85 patients were enrolled in this study and retrospective analysis of prognostic factors were compared with immunohistochemical staining of Bcl-2 and CD-44. RESULTS: The expression of Bcl-2 was relatively even when we compared with stage, cell types, grade and lymph node metastasis. And the CD-44 spliced variant form V6 expression was significantly strong in higher stage and positive lymph node metastasis but cell types and grade of tumor does not correlated with status of CD-44 expression. CONCLUSION: Bcl-2 expression in cervical cancer does not correlate with clinical prognostic factors, but CD-44 spliced variant V6 form does correlated with poor prognostic factors. CD-44 may play a role in the process of the tumor metastasis and poor prognosis.
Carcinogenesis ; Genes, bcl-2 ; Humans ; Lymph Nodes ; Neoplasm Metastasis ; Prognosis ; Retrospective Studies ; Uterine Cervical Neoplasms*

OBJECTIVETo investigate the Bcl-2 protein and gene expression in diffuse large B-cell lymphoma (DLBCL), and analyze its correlation with immunosubtype and prognosis.

METHODSSeventy-three cases of DLBCL were performed immunohistochemistry analysis with a panel of antibodies CD3, CD10, CD20, Bcl-6, Bcl-2 and MUM-1, and classified into germinal center B-cell (GCB) type and non-GCB type. Fluorescence in situ hybridization (FISH) was employed to detect bcl-2 gene expression in 57 cases with chromosome translocation t (14;18).

RESULTSThe percentages of tumor cells expressed CD10, Bcl-6, MUM-1 and Bcl-2 were 15.1%, 38.4%, 71.2% and 79.2%, respectively. 16 cases (21.9%) were GCB type and the rest (78.1%) were non-GCB type. 16 of 57 cases (28.1%) were t (14; 18), including 5 of GCB type (31.2%) and 11 of non-GCB type (68.2%). The expression of Bcl-2 protein was correlated with immunological subtype (P = 0.035), but not with survival time (P = 0.253). Between the t(14;18) positive and negtive groupes, there was significant difference for survival time (P = 0.022), but no difference for immunological subtype (P = 0.340). There was no correlation between Bcl-2 protein and t(14;18).

CONCLUSIONSGCB type DBLBCL with expression of Bcl-2 protein had a poor prognosis. t(14; 18) positive BLBCL had poor prognosis. The expression of Bcl-2 protein and t(14; 18) are usually discordant.

Genes, bcl-2 ; Germinal Center ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; Prognosis

Genes, Reporter ; Genetic Vectors ; Luciferases ; genetics ; Transcription, Genetic ; bcl-2-Associated X Protein ; genetics

OBJECTIVETo assess the protection against cisplatin-induced ototoxicity by adenovirus-mediated overexpression of the bcl-2 gene in cultured spiral ganglion cells (SGC).

METHODSSGC from P3 rats were cultured in vitro and exposed to adenovirus vector carrying green fluorescent protein gene (Ad-GFP), followed by immunocytochemical analysis for expression of the neuron-specific marker Neurofilament 200 (NF200) and detection under laser scanning confocal fluorescence microscope. Then, SGC were transduced by Ad-bcl-2 and the expression of human bcl-2 protein was evaluated by Western Blot. Finally, the cultures of SGC were divided into 4 groups: the group of Ad-bcl-2 transfection followed by cisplatin treatment, the group of Ad-GFP transfection followed by cisplatin treatment, the group of cisplatin treatment only and the untreated group. Cisplatin worked for 48 hours at a concentration of 2 microg/ml. Outcome measures included survival number of SGC and longest neurite length by using ImageJ software.

RESULTSSGC were cultured successfully in vitro and transfected by adenovirus vector safely and efficiently. By Western Blot, human bcl-2 protein was expressed in the group after exposure to Ad-bcl-2, but not in the Ad-GFP transfected SGC. Cisplatin exposure resulted in shrinking of neuritis and pyknosis of cell body, even cell death. Expression of bcl-2 in the SGC provided a significant level of protection against cisplatin-induced SGC degeneration.

CONCLUSIONSOur results suggest that SGC can be transduced by adenovirus vector safely and efficiently in vitro. Adenovirus-mediated delivery of the bcl-2 gene attenuates cisplatin-induced SGC degeneration.

Adenoviridae ; genetics ; Animals ; Apoptosis ; drug effects ; Cisplatin ; pharmacology ; Genes, bcl-2 ; Humans ; Spiral Ganglion ; cytology

We studied thirty benign and twenty-one malignant brain tumors in order to investigate the relationship between p53, bcl-2, apoptosis and histologic grade of brain tumors. For the study of p53 and bcl-2 gene expression, we used immunohistochemical staining method using monoclonal antibodies to p53 and bcl-2; and, for apoptosis, In-situ end labeling technique was used. The malignant group showed significantly higher p53 and apoptosis positive index(PI) than the benign group(mean p53 PI, malignant: 16.0 benign: 0.9/mean apoptosis PI, malignant: 2.3 benign: 0.2)(p=0.003); but bcl-2 positive index was not significantly different between two groups (p=0.118). Correlation between p53 mutation and apoptosis PI was statistically significant(p=0.012, Pearson coefficient=0.349); but correlation between bcl-2 expression and apoptosis PI was not(p=0.318). Moreover, correlation between p53 mutation and bcl-2 expression was not statistically significant(p=0.583). These results suggest that higher p53 mutation tends to exist in the group of tumors with higher malignant histologic grades. Furthermore, it can be concluded that greater DNA damage reflected by higher frequency of apoptosis tends to exist in the group of higher malignant histologic grade.
Antibodies, Monoclonal ; Apoptosis* ; Brain Neoplasms* ; Brain* ; DNA Damage ; Genes, bcl-2

PURPOSE: Alteration of p53 tumor suppressor genes is most frequently identified in human neoplasms, including urinary bladder tumor. The overexpression of the bcl-2 gene has been correlated with poor prognosis and chemotherapy resistance in other systems. We evaluated the significance of these gene expressions and correlation with prognostic factors in the urinary bladdder cancer. MATERIALS AND METHODS: The expression of bcl-2 and p53 oncoprotein were investigated serially in formalin-fixed and paraffin-embedded tissue specimens from 43 patients with primary transitioanl cell carcinoma in the urinary bladder. Thirty four were superficial bladder tumors and nine were invasive tumors. In histology grade according to WHO grading there were 9 grade I, 21 grade II and 13 grade III. RESULTS: Positive immunoreaction for bcl-2 was found in 3 out of 9 invasive bladder tumors, while 15 of 34 superficial tumors showed positive staining. Positive p53 immunostaining was found in 7 of 9 invasive tumors, while 18 of 34 superficial tumors showed positive staining. There was no significant correlation between bcl-2 or p53 expression and tumor stage(p=0.56, p=0.179). A higher incidence of bcl-2 staining was found in the grade I group than in the grade III group(p=0.041). There was no significant correlation between p53 expression and tumor grade (p=0.23). A significant inverse relationship was found between bcl-2 and p53 topographic expression(p=0.001). CONCLUSIONS: The results of this study indicated an inverse relationship between bcl-2 and p53 expression. These findings suggest bcl-2 expression may be associated with favorable prognosis in bladder tumor.
Drug Therapy ; Gene Expression ; Genes, bcl-2 ; Genes, Tumor Suppressor ; Humans ; Incidence ; Prognosis ; Urinary Bladder Neoplasms ; Urinary Bladder*