Role of Immunohistochemistry and serology in subclassifying the Inflammatory Bowel Disease cases diagnosed as Inflammatory Bowel Diseases-unclassified on colonic biopsies

Background: Inflammatory bowel diseases (IBDs) mainly consist of Crohn's Disease (CD) and Ulcerative Colitis (UC). These two categories have overlapping histopathological features and sometimes it is difficult to diagnose them into distinct category and such biopsies are categorised as Inflammatory Bowel Disease (IBD-U). Recently, there has been an increase in interest to discover new biomarkers of IBD to differentiate UC and CD and predict their prognosis. Method: In the present study, 273 non-neoplastic colonic biopsies with clinicoendoscopic features of IBD were studied and categorized into UC (88; 32.3%) and CD (03; 1.1%) but a major chunk remained in category of IBD-U (182; 66.6%). 161 (58.9%) of these biopsies were then subjected to IHC for RB protein and ?-catenin and Serology for pANCA and ASCA was done in only 85 (31.13%) of these selected cases for identification of UC and CD on colonic biopsies. Result: 161 biopsies that were subjected to IHC analysis included 57 cases of UC, 03 cases of CD, and rest 101 cases of IBD-U. Out of 101 cases of IBD-U, 87 (86.13%) cases were reclassified as UC (61; 60.3%) and CD (14; 13.86%) on the basis of results of IHC and Serology. Conclusion: The two major tools IHC for ?-catenin and RB protein and the assay of serum ASCA and p-ANCA along with proper history and clinical presentation can act as a good adjunct to conventional H and E in subclassifying cases of IBD-U into UC and CD.

Correlation analysis of human papilloma virus 16/18 infection and the expression of Rb and p16 protein in bladder cancer tissue / 国际检验医学杂志

Objective To investigate the correlation between human papilloma virus(HPV) 16/18 infection and the expression of Rb and p16 protein in bladder cancer tissue,and to analyze the relationship between HPV infection and the incidence of bladder cancer.Methods The expression of HPV16/18 E6 and E7 gene encoded protein,RB and p16 were detected by immunohistochemical method in 40 cases of bladder cancer and 40 cases of normal bladder tissues,and the correlation between them and pathological grading,stage of international union of cancer(UICC),whether recurrence or not after receiving surgery was analyzed.Results In bladder cancer tissues,HPV16/18 E6 and E7 gene encoded protein,RB and p16 positive rates were 65%,47.5%,42.5%,compared with the positive rate of normal bladder tissue samples(22.5%,92.5%,87.5%),the differences were statistically significant(P0.05).The expression of HPV16/18 E6 and E7 gene encoded protein and Rb,p16 protein were not significantly correlated(P>0.05),The expression of Rb and p16 protein were negatively correlated(P<0.05).Conclusion HPV 16/18 infection is related to the occurrence and development of bladder cancer,but its mechanism might not be related to the abnormal expression of Rb and p16 protein.

Correlation between p53 and Rb Protein Expression and Clinicopathologic Features in Hepatocellular Carcinoma / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Hepatocellular carcinomas (HCC) are one of the most common cancers in Korea. The mechanism of HCC development is still unclear, and the aberration of the tumor suppressor genes in HCC remains to be clarified. MATERIALS AND METHODS: To study the expressions of p53 and Rb protein, and their correlation with the clinicopathological parameters in HCC, 68 patients, with surgically resected hepatocellular carcinomas, were analyzed by an immunohistochemical method. The expressions of p53 and Rb protein were classified into three categorizes, depending on the percentage of stained cells. RESULTS: The expression of the p53 protein was 51.5% (35/68), and was significantly correlated with differentiation (p0.05). There was a positive correlation between p53 and Rb protein overexpression (p<0.05). The expressions of p53 and Rb protein had correlation with the Ki-67 labeling index (p<0.05). CONCLUSION: These findings suggest the aberrant expressions of p53 and Rb protein may play a role in the progression and carcinogenesis of HCC.

The role of dephosphorylated RB protein in human breast cancer cell apoptosis / 医学研究生学报

Objective: To investigate the role of the expression of dephosphorylated RB protein for cell apoptosis in human breast cancer. Methods: In our works, human breast cell lines (MCF-7/S,the chemosensitive cell line and MCF-7/Adr, the chemoresistent cell line)were evaluated. The expressive levels of dephosphorylated RB protein were detected with immunocytochemistry. Apoptosis rates were determined by flow cytometry(FCM). Results: Before treatment of ADR, phosphorylated RB protein was positive in two cell lines, but dephosphorylated RB protein was negative; After treatment of different concentration ADR, when the concentration of ADR was increased, expression of dephosphorylated RB protein elevated accordingly in MCF-7/S, but no significant change in MCF-7/Adr. Apoptosis and cycle was detected by FCM assays shows ADR induced apoptosis of MCF-7/S more than MCF-7/Adr(P

Altered Retinoblastoma Protein Expression and Proliferative Activity in Urethane Induced Mouse Lung Tumorigenesis / Journal of the Korean Cancer Association, 대한암학회지

Lung cancer develops through a multistage process involving the accumulation of diverse genetic alterations. To gain an understanding of the roles played by tumor suppressor gene proteins and proliferating cell nuclear antigen (PCNA) in chemical carcinogen-induced mouse lung tumorigenesis, we examined the expression of retinoblastoma protein (Rb), p53, and PCNA in normal lung tissues and urethane-induced mouse lung tumors. MATERIALS AND METHODS: ICR mice were given urethane by intra-peritoneal injection, and sacrificed at 5, 13, 21, 31, and 37 weeks following treatment. Sequential morphological changes and the immunohistochemical expression of Rb protein, p53, and (PCNA), during mouse lung tumorigenesis, were examined. RESULTS: During the carcinogenesis, sequential histological changes from hyperplasia of type II pneumocytes, to adenomas, and ultimately to overt adenocarcinomas were noted. Intense nuclear staining of the Rb protein was observed in normal and hyperplastic alveolar epithelial cells and adenomas. In adenocarcinomas, the Rb protein expression was significantly diminished. The p53 mutant protein was not detected in any lesion. The PCNA labeling index increased along with the advance in the histological grade. CONCLUSION: The above results indicate that mouse pulmonary adenocarcinomas develop through premalignant lesions, and down-regulation of the Rb protein expression may be implicated in the urethane-induced mouse lung tumorigenesis. In addition, the PCNA labeling index may reflect the malignant potential during the tumor progression.

Expression of G1/S Phase Checkpoint Proteins in Breast Carcinoma: Relationship to Clinicopathologic Factors andSurvival Rate / Journal of the Korean Cancer Association, 대한암학회지

The retinoblastoma protein (pRb)/cyclin D1/ p16 pathway plays a critical role in controlling the progression from G1 to S phase of the cell cycle. Abnormal expression of the individual components of the pathway has been reported in many human cancers, including the breast. Our aim was to investigate the role of this pathway in tumorigenesis and tumor progression, and to evaluate the value of these oncoproteins as potential prognostic factors in breast cancer. MATERIALS AND METHODS: We examined the significance of the p16, pRb, and cyclin D1 expression in 128 cases of invasive breast carcinomas using immunohistochemistry on formalin fixed, paraffin sections. The results correlated with the survival rate and clinicopathologic variables, including age, histologic grade, lymph node status, tumor size, estrogen receptor (ER) and progesterone receptor (PR) content. The negative finding for nuclear staining for pRb and p16 were defined as abnormal. RESULTS: Abnormal expression of the p16 and pRb were seen in 21% and 43% of tumors, respectively. There was a significant inverse relationship between the p16 and pRb expressions. There was no association between the p16 staining and any other parameters, including survival rate, cyclin D1, or clinicopathologic variables. Surprisingly, there was a trend for pRb positive tumors to be grade III ductal carcinomas. Cyclin D1 positivity was noted in 46% of cases. The expression of cyclin D1 protein was significantly higher in lower histologic grades, and with higher ER and PR expressions. CONCLUSION: These findings suggest the p16 may be negatively regulated by the pRb, and that cyclin D1 is involved in the tumor progression in well-differentiated tumors and could be an ER and PR related protein. In a Cox multivariate analysis, the p16, pRb, and cyclin D1 were not independent predictors of patient outcome.

The Effect of Rb Protein Expression on the Clinical Response of the Colorectal Cancer Patients Perfused with 5-Fluorouracil Before Operation / 中国医师杂志

Objective To investigate the effect of Rb protein expression on the clinical response of the colorectal cancer patients perfused with 5-Fu before operation. Methods The expression of Rb protein in 34 cases of the colorectal cancer sample was determined by streptavidin-peroxidase(S-P) immunohistochemical method and the positive reactions were analysed by ICM-100 Cell DNA Picture Analysis System. The apoptotic rate of colorectal cancer cells was examined by the TUNEL staining. The chemosensistivity of the colorectal cancer cells were tested by the MTT colorimetric assay of agar culture. The clinical response of the colorectal cancer patients perfused with 5-Fu before operation was observed simultaneously. Results ⑴There was the negative correlation between the resistance index of the colorectal cancer cells to 5-Fu (IC50/10-1 peak concentration) and the apoptotic rate of the colorectal cancer cells induced by 5-Fu (?=-0.53,t=-4.91,P1.0 belongs to the group of resistance,

Apoptosis in Renal Cell Carcinoma: Correlation to Apoptosis Related Genes and Cell Proliferation, and Its Prognostic Significance / 대한암학회지

PURPOSE: To investigate the prognostic role of apoptosis and to evaluate the relationship between apoptosis and apoptosis-related genes, as well as cell proliferation in renal cell carcinoma (RCC). MATERIALS AND METHODS: Apoptosis was detected by using the terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) technique in 67 formalin-fixed and paraffin-embedded RCC specimens. Immunohistochemical stainings for p53 and retinoblastoma (Rb) proteins and proliferating cell nuclear antigen (PCNA) were also conducted simultaneously. RESULTS: The apoptotic index (AI) varied from 0.2% to 25.5%. The PCNA index (PI) ranged from 2.1% to 70.3%. The expression of p53 protein was found in 31 of 67 (46.3%) cases. Abnormal expression of Rb was seen in 23 of 67 (34.3%) cases. There was a statistically significant positive correlation between AI and increasingnuclear grade (p<0.001). A significant correlation was found between AI and PI (r=0.329, p<0.01). When comparing the AI with the expression of p53 and Rb proteins, there was no significant difference. In univariate survival analysis, nuclear grade, TNM stage, PI, expression of Rb and AI were significantly associated with shortened survival. However, TNM stage was the only independent prognostic factors by multivariate analysis. CONCLUSION: The present findings indicate that apoptosis in RCC is closely associated with cell proliferation, but not with the expression of p53 and Rb proteins. In multivariate analysis, the AI does not carry an independent prognostic significance.

A Study on Expression Pattern of p53, Rb Gene and Apoptosis in Ovarian Epithelial Borderline Tumors and Invasive Carcinoma / 대한산부인과학회잡지

OBJECTIVE: The aim of this study is to evaluate the role of tumor suppressor genes, p53 and Rb gene, as well as apoptosis in the carcinogenesis of ovarian epithelial tumors. And the value of these factors as prognostic markers to tell the transformation of borderline tumors to overt carcinomas is also studied. METHOD: Thirty cases of ovarian epithelial benign and borderline tumors and invasive carcinoma were used and the expression of the p53 protein and Rb gene protein were evaluated by immunohistochemical method. The apoptosis was evaluated by TUNNEL method. RESULTS: Positive rate of p53 expression in benign, borderline and invasive tumors were 0, 28, and 94 %, respectively. And also, p53 was highly expressed in chemoresistant cases (2/3), in residual tumor (4/5) and in recurred cancer (2/2). Rb protein was partly lost in the borderline tumors, but the rate of Rb protein loss in both borderline tumors and invasive carcinomas were similar. Apoptosis were more active in overt carcinomas than in borderline and benign tumors. In borderline tumors, p53 protein was expressed as 28.6% positivity, and apoptosis was expressed as 28.6% negativity, which showed indirectly that there was apoptosis induction effect of p53. In ten cases of invasive carcinomas showing highly expressed p53, apoptosis revealed all positive reaction except 2 cases, and Rb protein revealed variously. This result supported the apoptosis imduction effect of p53, but it was difficult to find the association of expression degree between the two tumor supressor genes CONCLUSION: In conclusion, the values of p53 is a discriminating factor of malignancy from benign and the expression of p53 is related with clinical aggressivity such as recurrence and residual cancers. Apoptosis are more active in overt carcinoma than in benign & borderline tumor, and in borderline tumor the expression of p53 is related to apoptosis induction which results to carcinomatous change.

Expression of p53 and Rb Proteins in Invasive Ductal Carcinoma of the Breast

Inactivation of tumor suppressor genes may play an important role in many human cancers including breast. This study was done to determine the relationship between the expression of p53 and Rb protein and prognostic factors such as histopathologic differentiation, tumor size, and lymph node metastasis. In 57 cases of breast invasive ductal carcinomas, the immunohistochemical staining with p53 and Rb protein gave the following results: p53 protein was detected in 45.6% (26/57) of cases. Tumors with large size, poor differentiation or lymph node metastases tended to show increased expression of p53 protein. However, p53 protein expression did not show any significant correlation with prognostic factors such as tumor size (p value 0.25), histologic grade (p value 0.75), and positive lymph node status (p value 0.26). Rb protein was detected in 57.9% (33/57) of cases. Rb protein also did not show any significant correlation with prognostic factors such as tumor size (p value 0.56), histologic grade (p value 0.71), and positive lymph node status (p value 0.98). There was no significant correlation between p53 expression and Rb protein expression (p value 0.80).

Relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer / 中国癌症杂志

Purpose:To investigate the relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer. Methods:In our work,human breast cell lines (MCF-7/S,the chemosensitive cell line and MCF-7/ADR,the chemoresistent cell line)were evaluated. Chemosensitivity of two cell lines was evaluated by the MTT colorimetric assay;the expressive levels of dephosphorylated RB protein were detected with immunocytochemistry. Apoptosis rates were determined by flow cytometry(FCM). Results:ADR inhibited proliferation of chemosensitive cell line MCF-7/S ,the 50% inhibition concentration (IC 50 ) was 0.128 ?g/ml;And IC 50 of MCF-7/ADR was 10.89 ?g/ml. The chemotherapeutic sensitivity of MCF-7/S was more than that of MCF-7/ADR by 86 times . Before treatment with ADR,phosphorylated RB protein was positive in two cell lines,but dephosphorylated RB protein was negative;After treatment of different concentration ADR,when the concentration of ADR was increased,expression of dephosphorylated RB protein elevated accordingly in MCF-7/S,but no significant change in MCF-7/ADR. Apoptosis and cell cycle was detected by FCM assays shows ADR induced apoptosis of MCF-7/S more than MCF-7/ADR(P0.05).

Expression Pattern of the Rb Protein and its Correlation with Prognosis in Primary Lung Cancer

An immunohistochemical stain for the Rb tumor suppressor gene product was performed in pathologic specimens from 72 primary lung cancer patients to study the correlation between its expression and histologic type, cancer differentiation, clinical stage and survival rate. The expression of the Rb protein was positive in 34 cases(47.2%) and negative in 38 cases(52.8%). The Rb protein was not expressed in 16 of 42 cases(38.1%) in squamous cell carcinoma, in 17 of 23 cases(73.9%) in adenocarcinoma, in one of three cases(33.3%) in undifferentiated large cell carcinoma, in two of two cases(100%) in small cell carcinoma, in one of one case(100%) in an adenosquamous carcinoma and in one of one case(100%) in an atypical carcinoid. There were significant difference of the Rb protein expression between squamous cell carcinoma and adenocarcinoma(p0.05). The two year survival rate for patients with the Rb positive was 65% compared with 37% for those with the Rb negative which was significant(p<0.05). This result suggests that an altered or the absence of the Rb protein in cancer cells can be a valuable prognostic factor in the lung cancer.

Expression Pattern of the Rb Protein and its Correlation with Prognosis in Primary Lung Cancer

An immunohistochemical stain for the Rb tumor suppressor gene product was performed in pathologic specimens from 72 primary lung cancer patients to study the correlation between its expression and histologic type, cancer differentiation, clinical stage and survival rate. The expression of the Rb protein was positive in 34 cases(47.2%) and negative in 38 cases(52.8%). The Rb protein was not expressed in 16 of 42 cases(38.1%) in squamous cell carcinoma, in 17 of 23 cases(73.9%) in adenocarcinoma, in one of three cases(33.3%) in undifferentiated large cell carcinoma, in two of two cases(100%) in small cell carcinoma, in one of one case(100%) in an adenosquamous carcinoma and in one of one case(100%) in an atypical carcinoid. There were significant difference of the Rb protein expression between squamous cell carcinoma and adenocarcinoma(p0.05). The two year survival rate for patients with the Rb positive was 65% compared with 37% for those with the Rb negative which was significant(p<0.05). This result suggests that an altered or the absence of the Rb protein in cancer cells can be a valuable prognostic factor in the lung cancer.

Enhancement of cytotoxicity of Adramycin by neferine in Saos-2 cells and its mechanism / 中国药理学通报

AIM To study the synergistic anticancer effccts of neferine combined with anticancer agents on Saos 2 cell lines and its mechanism. METHODS\ Cytotoxicity assay was tested by MTT method.Cell cycle and apoptosis was detected by PI staining flow cytometty. Expression of Rb protein was examined by indirect immune fluorescent symbol flow cytometty and S P immunohistochemical staining. RESULTS\ 5,10 ?mol?L -1 Nef augmented apoptosis of Saos 2 cells induced by ADM,enhanced the expression of Rb protein in Saos 2 cells and raised the G 1 phase ratio of Saos 2 cells. CONCLUSION Nef can promote chemotherapy sensitivity to ADM in Saos 2 cells. Its mechanism is associatd with the up modulation of Rb protein expression in the Saos 2 cells arrest cells in G 1 phase

Expression of cyclin D1 and Rb protein in breast cancer / 中国免疫学杂志

Objective:To detect the expression of cyclin D1 protein and Rb protein in breast cancer and analyze its relationship to carcinogenesis and development.Methods:The expression of cyclin D1 and Rb protein were detected in 52 breast cancer and 20 benige breast tissues by S-P immunohistochemical method.Results:The positivity rate of cyclin D1 overexpression in the breast cancer was 34.6%(18/52),while there was a significant difference compared with that of benign breast tissues 10%(2/20),P