Masoud Hassanzadeh Makoui; Maryam Mobini; Jalal Khoshnoodi; Tannaz Bahadori; Forough Golsaz-Shirazi; Hedieh Moradi Tabriz; Zahra Madjd; Mahmood Jeddi-Tehrani; Amir-Hassan Zarnani; Mohammad Mehdi Amiri; Fazel Shokri
Abstract
Background: Ki67 and P53 are important diagnostic and prognostic biomarkers expressed in several cancers. The current standard method for evaluating Ki67 and P53 in cancer tissues is immunohistochemistry (IHC), and having highly sensitive monoclonal antibodies against these biomarkers is necessary for ...
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Background: Ki67 and P53 are important diagnostic and prognostic biomarkers expressed in several cancers. The current standard method for evaluating Ki67 and P53 in cancer tissues is immunohistochemistry (IHC), and having highly sensitive monoclonal antibodies against these biomarkers is necessary for an accurate diagnosis in the IHC test.Objective: To generate and characterize novel monoclonal antibodies (mAbs) against human Ki67 and P53 antigens for IHC purposes.Methods: Ki67 and P53-specific mAbs were produced by the hybridoma method and screened by enzyme-linked immunosorbent assay (ELISA) and IHC techniques. Selected mAbs were characterized using Western blot and flow cytometry, and their affinities and isotypes were determined by ELISA. Moreover, using the IHC technique in 200 breast cancer tissue samples, we assessed the specificity, sensitivity, and accuracy of the produced mAbs.Results: Two anti-Ki67 (2C2 and 2H1) and three anti-P53 mAbs (2A6, 2G4, and 1G10) showed strong reactivity to their target antigens in IHC. The selected mAbs were also able to recognize their targets by flow cytometry as well as Western blotting using human tumor cell lines expressing these antigens. The specificity, sensitivity, and accuracy calculated for clone 2H1 were 94.2%, 99.0%, and 96.6%, and for clone 2A6 were 97.3%, 98.1%, and 97.5%, respectively. Using these two monoclonal antibodies, we found a significant correlation between Ki67 and P53 overexpression and lymph node metastasis in patients with breast cancer.Conclusion: The present study showed that the novel anti-Ki67 and anti-P53 mAbs could recognize their respective antigens with high specificity and sensitivity and therefore can be used in prognostic studies.
Nasser Gholijani; Ahmad Monabati; Zahra Amirghofran
Volume 1, Issue 1 , June 2004, , Pages 34-40
Abstract
Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer ...
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Background: Prostate cancer is one of the most commonly diagnosed cancers in males. Tumor suppressor gene p53 plays an important role in causing cell cycle arrest and allowing apoptosis to proceed. Objective: To investigate the expression of p53 protein and its relation to apoptosis and prostate cancer traditional prognostic indicators. Methods: In this study expression of p53 was examined in paraffin-embedded tissues from 50 cases of prostate carcinoma by immunohistochemistry and evaluated using an index of staining. Correlation between p53 expression and apoptosis was detected by TUNEL method. Pathological grade, Gleason score and stage of carcinoma were also determined. Results: P53 expression was observed in 48 of 50 cases (26-100% of tumor cells) with mean staining index of 141±65. A significant association between p53 expression and pathologic grade (r=0.37, p=0.004) and Gleason score (r= 0.4, p=0.009) of patients was observed. Apoptosis was detected in only 6 patients. p53 expression showed no correlation with apoptotic index. No correlation between p53 expression and stage or apoptosis and clinicopathological characteristics of patients was found. Conclusion: p53 expression showed a significant correlation with differentiation status of the prostate carcinoma and can be helpful as a prognostic marker. Decreased level of apoptosis observed in our cases was not correlated with p53 expression indicating the possible role of other regulatory molecules involved in the apoptosis.