Immunohistochemical Evaluation Of Ki-67 Research Articles

Exploring the osteogenic potential of semisynthetic triterpenes from Combretum leprosum: An in vitro and in silico study.

Combretum leprosum Mart. is a plant of the Combretaceae family, widely distributed in the Northeast region of Brazil, popularly used as an anti-inflammatory agent, and rich in triterpenes. This study evaluated in vitro and in silico potential osteogenic of two semisynthetic triterpenes (CL-P2 and CL-P2A) obtained from the pentacyclic triterpene 3β,6β,16β-trihydroxylup-20(29)-ene (CL-1) isolated from C. leprosum. Assays were carried out in cultured murine osteoblasts (OFCOL II), first investigating the possible toxicity of the compounds on these cells through viability assays (MTT). Cell proliferation and activation were investigated by immunohistochemical evaluation of Ki-67, bone alkaline phosphatase (ALP) activity, and mineralization test by Von Kossa. Molecular docking analysis was performed to predict the binding affinity of CL-P2 and CL-P2A to target proteins involved in the regulation of osteogenesis, including: bone morphogenetic protein 2 (BMP-2), proteins related to Wingless-related integration (WNT) pathway (Low-density lipoprotein receptor-related protein 6-LRP6 and sclerostin-SOST), and receptor activator of nuclear factor (NF)-kB-ligand (RANK-L). Next, Western Blot and immunofluorescence investigated BMP-2, WNT, RANK-L, and OPG protein expressions in cultured murine osteoblasts (OFCOL II). None of the CL-P2 and CL-P2A concentrations were toxic to osteoblasts. Increased cell proliferation, ALP activity, and bone mineralization were observed. Molecular docking assays demonstrated interactions with BMP-2, LRP6, SOST, and RANK-L/OPG. There was observed increased expression of BMP-2, WNT, and RANK-L/OPG proteins. These results suggest, for the first time, the osteogenic potential of CL-P2 and CL-P2A.

The expression of Ki-67 marker in the hydatidiform mole

Background: The hydatiform mole is charcaterized by a pathological proliferation of the trophoblast. Its evaluation could predict the progression to gestational trophoblastic neoplasia. The aim of the study was the analysis of the proliferative activity of the villous trophoblast in the molar vs non-molar lesions. Material and methods: p-57 and Ki-67 were evaluated by immunohistochemistry in 15 cases of hydatidiform mole and 18 cases of abortions. Results: The hydatidiform mole was divided based on the immunoexpression of anti-p57 into: complete (8 cases) and partial type (7 cases). The distribution score of Ki67 immunoreactivity in the villous cytotrophoblast was: complete mole: +3 – 8 cases; partial mole: +1 – 1 case, +2 – 3 cases, +3 – 3 cases; abortions on social indications: +1 – 6 cases, +2 – 9 cases; +3 – 2 cases. The mean and standard deviations were: 2.88±0.354; 2.29±0.756 and 1.82±0.728, respectively. The following statistical correlations were determined: complete vs partial mole (p=0.014), complete mole vs abortion (p<0.01) and overall cases of mole vs abortion (p=0.000034). Conclusions: Villous cytotrophoblast proliferative activity is high in the complete hydatidiform mole, and the immunoreactivity distribution index is highly positive and statistically true in the molar versus non-molar group. Immunohistochemical evaluation of Ki-67 in molar pathology is useful in differential diagnosis as a complementary method.

HISTOPATHOLOGICAL STUDY OF SOFT TISSUE TUMOURS AND CORRELATION OF HISTOLOGIC GRADE OF SARCOMAS WITH PROLIFERATIVE MARKER KI-67

INTRODUCTION:Soft tissue tumours have diagnostic and therapeutic challenge due to a wide variety of histomorphological patterns. Soft tissue sarcomas are relatively infrequent and constitute less than 1 % of all malignant neoplasms. Immunohistochemical evaluation of Ki-67 is a rapid prognostic tool to assess the cellular proliferation better than mitotic index. METHODS: A prospective study was conducted with a total of 100 cases of soft tissue tumors to analyze the frequency and histomorphology. Soft tissue sarcomas were graded by FNCLCC grading system and were subjected to Ki-67 evaluation. Spearman's Rho and Pearman's correlation coefcients were calculated to determine the correlation between the histologic grade and Ki-67 index. RESULTS: Soft tissue tumours constituted only about 1.3% of all neoplasms. Benign tumours (50%) outnumbered intermediate (11%) and malignant soft tissue tumours (39%). Soft tissue tumours were more common in males (52%) than females (48%) and frequently observed in the sixth decade. The sites of predilection were upper extremities for benign tumours and lower extremities for malignant tumours. Most common benign soft tissue tumour was Neurobroma (19%) followed by Schwannoma (14%). Undifferentiated pleomorphic sarcomas were the most common malignant soft tissue tumour (28%). In FNCLCC grading, most of the sarcomas were grade III (42%). Ki-67 index was low in grade I sarcomas, variable in grade II sarcomas and high in grade III sarcomas. Spearman's rho and Pearman's correlation R values were 0.77 and 0.75 respectively and p value is less than 0.05. CONCLUSION: There is positive correlation between the histologic grade and Ki-67 proliferative index in soft tissue sarcomas. Ki-67 index can be used as an independent prognostic factor to predict the risk of distant metastasis. Prospective evaluation of Ki-67 should be carried out in patients with soft tissue sarcomas for planning adjuvant treatment modalities

Comparative Analysis of Ki-67 in Different Scoring Patterns and its Association with other Prognostic Markers of Breast Carcinoma

Introduction: Immunohistochemical evaluation of Ki-67 is widely used for estimation of tumour proliferation in breast cancer. Till date, no specific method or a cut-off point for Ki-67 exists. Aim: To perform a comparative analysis between different scoring patterns and mean Ki-67 value and association of mean Ki-67 with other prognostic markers like tumour size, lymph node status, tumour stage and grade and different molecular subtypes of breast cancer. Materials and Methods: A cross-sectional study was conducted at Dr. D.Y Patil Medical College, Hospital and Research Centre, Pune, Maharashtra, India, between August 2019 to August 2021. Total of 50 new diagnosed cases of breast cancer were studied for the histologic type, grade and stage of the tumour. Immunohistochemistry for Oestrogen Receptor (ER), Progesterone Receptor (PR), Human Epidermal Growth Factor Receptor 2 (HER2) and Ki-67 was performed. Association of Ki-67 with other prognostic markers like tumour size, lymph node status, tumour stage and grade and different molecular subtypes of breast cancer was evaluated by expressing Ki-67 as a continuous variable (mean±SD) and also by dividing Ki-67 into different scoring patterns (I: ≤14%, >14%, II: ≤15%, 16- 30%, >30% and III: <20%, 20-50%, ≥50%). Statistical tests like Kruskal-Wallis test (mean Ki-67 with tumour size, tumour grade and molecular subtypes), Mann-Whitney Rank Sum Test (mean Ki-67 with lymph node status) and Analysis of variance (ANOVA) (mean Ki-67 with staging) respectively. Results: Out of 50 patients, 40 (80%) were older than 50-yearold. Twenty six (52%) cases affected the left breast. A total of 49 (98%) were diagnosed as Invasive Ductal Carcinoma (IDC). Among them 26 (52%) cases were of grade III and 25 (50%) cases were of Luminal A. Mean Ki-67 and molecular subtypes of breast cancer had statistically significant association (p= 0.002). No association was found between mean Ki-67 and tumour size (p=0.608), lymph node status (p=0.506) stage (p=0.979) and grade (p=0.095) of the tumour. Although scoring pattern I and III had no remarkable findings. Notably, scoring pattern II showed higher tumour sizes, lymph node positivity, higher stage and grade and basal-like tumours demonstrated a higher Ki-67 index. Conclusion: Evaluation of Ki-67 as a continuous variable yielded significant association with other prognostic markers of breast cancer. There was no single “best” scoring pattern identified. A direct association of Ki-67 was found with molecular subtypes of breast cancer.

Immunohistochemical evaluation of Ki-67, Cyclin D1 and β-catenin expression in the subtypes of triple negative breast cancer

Aim. To evaluate the expression levels of Ki-67 and cyclin D1 and β-catenin in the subtypes of triple negative breast cancer. Methods. The study was conducted on the surgical material from 60 patients of clinical stage 2A (T1N1M0 or T2N0M0) who were treated at the Rostov Research Institute of Oncology from 2012 to 2015. For immunohistochemistry, antibodies to estrogen and progesterone receptors, cytokeratins 5/6, Ki-67, cyclin D1, β-catenin, HER2/neu and EGFR proteins were used. Results. Triple negative breast cancer with the signs of basal epithelium was found to have a significantly higher expression level of Ki-67 compared to non-basal-like one. In some part of triple negative breast cancer samples overexpression of cyclin D1 was observed. The high level of cyclin D1 in the basal-like subtype was less common than in the subtypes without the signs of basal epithelium, but its average value was significantly higher. In triple negative cancer with cyclin D1 overexpression, the loss of β-catenin on the cell membrane and its abnormal accumulation in the cytoplasm was significantly more frequent. β-catenin translocation into the cell nucleus was observed only in basal-like triple negative cancer, and 2 times more often in case of cyclin D1 overexpression. Conclusion. In triple negative breast cancer tumors with overexpression of cyclin D1 and abnormal expression of β-catenin are observed in some cases; these biomarkers can be considered as potential therapeutic targets for this group of tumors.

Immunohistochemical Evaluation of Ki-67 and Comparison with Clinicopathologic Factors in Breast Carcinomas

Background:Patients primarily received tamoxifen based on their menopausal status due to the lack of immunohistochemistry. A recent study has shown that hormonal receptors were not correlated with menopausal status, and thus, indicating that they present limited therapeutic and prognostic significance in breast cancer management. This study aimed to evaluate Ki-67 value and analyze its association with clinicopathologic parameters in breast cancer patients.Methods:The formalin-fixed paraffin-embedded breast tissue blocks of 125 patients with primary breast carcinomas were subjected to immunohistochemical analysis using Ventana Benchmark® GX automated immunostainer. Analysis of variance and Chi-2 test were used to examine the relationship between Ki-67 and clinicopathologic variables.Results:The mean age of 125 patients included in the study was 47.7 years. The average score of Ki-67 was 56.0%. 84.8% of patients showed Ki-67 ≥ 14%. Mean scores of Ki-67 were correlated with grade (p = 0.006), PR (p = 0.026), histological type, ER, combined ER/RP, and molecular subtype (p < 0.001). Ki-67 was independent of HER2 (p = 0.402) and menopausal status (p = 0.471). The frequency of Ki-67 according to St Gallen 2011 was associated with histological type (p = 0.005), grade (p = 0.005), ER (p < 0.001), combined ER/PR (p = 0.004), and molecular subtype (p = 0.004). There was no significant relationship between the distribution of Ki-67 and the age of the patients (p = 0.859), menopausal status (p = 0.979), PR (p = 0.149), and HER2 (p = 0.597).Conclusion:Ki-67 is useful for treatment decisions in primary breast cancer patients. The high value of Ki-67 was associated with adverse clinicopathologic factors. The increased Ki-67 value should be carefully investigated in triple negative patients.

Ki67 as a Biomarker of Prognosis and Prediction: Is it Ready for Use in Routine Pathology Practice?

Breast cancer is the most common cancer in women, and the search for effective markers to design therapeutic strategies and patient management algorithms is still a work in progress. Ki67, a proliferative marker, has gained attention as a prognostic and predictive factor in early breast cancer and/or to decide response to chemotherapy. Individual studies and meta-analyses have provided evidence of its usefulness in this regard. Immunohistochemical staining Ki67 has emerged as an easy and cheap tool to assess proliferation in laboratory setting. However, debate continues over its meaningful clinical use, given the lack of standardization of staining techniques and firm recommendations about pre-analytical tissue handling, interpretation of the stain, and methods to estimate Ki67, resulting in high interlaboratory and interobserver variability. Importantly, no consensus has been reached on the cut-off values for risk stratification. The Breast Cancer Working Group proposed guidelines for immunohistochemical evaluation of Ki67 in 2011. However, the follow-up study showed poor reproducibility even among experts, and the use of Ki67 in daily practice is still in question.

Immunohistochemical Evaluation of Ki-67, MMP-9 and CD3+ Lymphocytes in Feline Mammary Cancer – Correlation with Classification Standards

Immunohistochemical Evaluation of Ki-67, MMP-9 and CD3+ Lymphocytes in Feline Mammary Cancer – Correlation with Classification Standards

Prognostic value of proliferation in pleomorphic soft tissue sarcomas: a new look at an old measure

Though proliferation has repeatedly shown a prognostic role in sarcomas, it has not reached clinical application. We performed a comprehensive evaluation of the prognostic role of 5 proliferation measures in a large series of soft tissue sarcomas of the extremities and the trunk wall. One hundred ninety-six primary soft tissue sarcomas of the extremities and the trunk wall were subjected to DNA flow cytometry for quantification of S-phase fraction and to immunohistochemical evaluation of Ki-67, Top2a, p21, and p27Kip1. In univariate analysis, positive expression of Ki-67 (hazard ratio = 4.5, CI = 1.6-12.1), Top2a (hazard ratio = 2.2, CI = 1.2-3.5) and high S-phase fraction (hazard ratio = 1.8, CI = 1.2-3.7) significantly correlated with risk for metastasis. When combined with currently used prognostic factors, Ki-67, S-phase fraction and Top2a fraction contributed to refined identification of prognostic risk groups. Proliferation, as assessed by expression of Ki-67 and Top2a and evaluation of S-phase fraction and applied to statistical decision-tree models, provides prognostic information in soft tissue sarcomas of the extremity and trunk wall. Though proliferation contributes independently to currently applied prognosticators, its role is particularly strong when few other factors are available, which suggests a role in preoperative decision-making related to identification of high-risk individuals who would benefit from neoadjuvant therapy.

Clinicopathological features of salivary and non-salivary adenoid cystic carcinomas

Adenoid cystic carcinoma (ACC), commonly from salivary glands, is known for its insidious local growth and usually protracted clinical course. ACC developing from non-salivary glands (i.e., non-salivary ACC) is heterogeneous, and its clinicopathological features remain poorly defined. Patients treated for ACC in a single institution between 1995 and 2007 were included in this study. Immunohistochemical evaluation of Ki-67, E-cadherin, p16, and cyclinD1 was performed. The prognostic significance of clinical and immunophenotypic markers was evaluated. 83 cases of salivary ACC and 24 cases of non-salivary ACC were included. The expression levels of Ki-67 (54.8%), E-cadherin (90.4%), p16 (32.9%), and cyclinD1 (19.2%) between ACCs present at various sites were not different. Sinonasal, lacrimal, and tracheobronchial ACCs had significantly worse outcomes than those of ACC of the major salivary glands. Postoperative radiotherapy reduced the recurrence rate of patients with a negative resection margin (P=0.028). Older age (age >60 years), advanced stage, positive resection margin, high histological grade, and high expression of Ki-67 were significantly correlated with poor prognosis. In conclusion, the site of origin plays a role in the prognosis of ACC, in which positive resection margin and advanced stage are possible factors underlying the differences in outcomes.

Expression of Ki-67 as a proliferation marker in prostate cancer

Prostate carcinoma (PCa) is the most common male cancer found in industrialized societies and represents a serious public health problem. Delineation of gene expression patterns in early PCa that correlate with an aggressive phenotype is a priority and may allow for radical treatment to be offered on a more selective basis to those patients with a clinically localized, yet aggressive disease. Ki-67 was recognized as associated with carcinogenesis in PCa. The aim of this study was the immunohistochemical evaluation of Ki-67 and its expression in PCa following radical prostatectomy, and analysis of its relationship to chosen clinical and morphological parameters of such tumors. A total number of 56 randomly selected patients undergoing radical prostatectomy were investigated. The tumors, after fixation with 10% neutral buffered formalin, were completely embedded in paraffin. The sections were cut into hematoxylin-eosin staining for histological examination. The sections were also immunostained, with monoclonal antibodies against Ki-67. Immunolocalization of Ki-67 was performed using the LSAB method. Serum PSA levels were obtained from clinical information. These obtained results were statistically analyzed using the Fisher's exact test and χ 2 test. No statistically significant correlation was found between the expression of Ki-67 and the preoperative PSA level, lymph node metastases, capsular penetration, seminal vesicle invasion, and positive or negative surgical resection margins. However, a strong statistically significant correlation between Ki-67 positive and the T stage was found. We also found a relationship between the Gleason score of 7 or above and a high expression of Ki-67 in PCa (p < 0.004, p < 0.02 respectively). It is noteworthy that a significant correlation exists between the Gleason score and the expression of Ki-67 in this present study. We observed a high expression of Ki-67 for a Gleason score of 7 or above. Our results suggest that Ki-67 may be useful to serve as a tumor marker in PCa.

New trends in primary systemic therapy for breast cancer

Neoadjuvant chemotherapy (NAC) is the standard treatment for locally advanced breast cancer. NAC is also recognized as the initial treatment of choice for patients who are candidates for adjuvant chemotherapy. A pathologic complete response (pCR) is a predictor of outcome associated with improved disease-free and overall survival. The identification of accurate predictors of a pCR to NAC is important to spare patients unnecessary toxicity. Hormone receptors, histologic grade, HER2, and ki-67 labeling index were evaluated as predictors of a pCR. An early response to NAC might be correlated with a high probability of a pCR. Jinno et al. reviewed indications for and predictors of NAC. The histological effects differ among the subtypes of breast cancer when the same NAC regimen is administered. In clinical studies of patients with HER2positive breast cancer, NAC with trastuzumab resulted in high pCR rates. The induction of personalized medicine and new molecular targeted therapies are expected to result in higher pCR rates in NAC. Dr Kinoshita summarized the current consensus on the adoption and benefits of NAC, especially for operable breast cancer patients. Hormone receptor (HR)-positive, HER2-negative breast cancer is regarded as a subtype with a good prognosis and which is sensitive to hormone therapy. Endocrine therapy may be sufficient for some HR-positive, HER2-negative breast cancer patients. Neoadjuvant endocrine therapy (NAE) is a new treatment option for such patients. Takei et al. reviewed NAE for premenopausal breast cancer patients using a luteinizing hormone-releasing hormone (LH-RH) analogue plus tamoxifen and for postmenopausal breast cancer patients using tamoxifen or aromatase inhibitors. The prognosis of HR-positive breast cancer patients receiving NAE has not been evaluated thoroughly. No comparative study of pre and postoperative endocrine therapy is available. Dr Iwata has introduced a new clinical study, N-SAS BC06, which is a randomized phase III trial conducted in Japan. Postmenopausal women with ER-positive, HER2-negative T1c-T2 N0 M0 breast cancer were classified into complete response (CR), partial response (PR), or stable disease (SD) groups and the progressive disease (PD) group, and patients in the CR, PR, or SD groups were randomized to a chemotherapy followed by endocrine therapy group or to an endocrine therapy group. The primary endpoint is disease-free survival. Immunohistochemical (IHC) biomarkers such as ER, PgR, and HER2 are very important predictors of the sensitivity to endocrine therapy or chemotherapy. Recently, there has been a focus on IHC evaluation of Ki-67 as a predictor of breast cancer outcome. Kumaki et al. reviewed our present understanding of the IHC evaluation of biomarkers for breast cancer under NAC.

Immunohistochemical evaluation of Ki-67, PCNA and MCM2 proteins proliferation index (PI) in advanced gastric cancer.

The current study objective was to assess the proliferation indices (PI) of Ki-67, PCNA and MCM2 proteins in advanced gastric cancer and in metastatic lymph node in correlation with certain clinicopathological features and with postoperative survival of patients. The study was conducted in a group of 100 patients with advanced gastric cancers. Involvement of local lymph nodes was present in 36 cases. Immunohistochemical investigations were carried out using monoclonal antibodies against Ki-67 (DAKO), PCNA (DAKO) and polyclonal antibody to MCM2 (Santa Cruz Biotechnology). Visualization of the antigen/antibody complex was performed using LSAB technique (biotin-streptavidin-peroxidase) followed by application of chromogene DAB (DAKO). Statistical analysis revealed no correlations of Ki-67, PCNA and MCM2 PI in tumour tissue or metastatic lymph node with patients' age and gender, tumour location, histological grade, macroscopic type according to Bormann's classification and histological grading by Lauren's and Goseki's classifications. Moreover, no correlation was observed of Ki-67 and MCM2 PI in tumour tissue with histological grading. No correlation was also noted between the proliferation indices of all the three proteins in the affected lymph node and grade of histological differentiation. Such clinicopathological parameters as patients' age and gender, histological grading by Lauren's and Goseki's classifications and lymph node involvement did not correlate with survival time of patients. Furthermore, no statistically significant correlation was shown of postoperative survival time with Ki-67 and MCM2 PI in tumour tissue and metastatic lymph nodes and with PCNA PI in the affected lymph nodes. However, a statistically significant correlation was found of Ki-67, PCNA and MCM2 PI in tumour tissue and metastatic lymph nodes with depth of wall invasion and local lymph node involvement. A statistically significant correlation was also noted between PCNA PI in the main mass of tumour and histological grading. The postoperative survival time of patients exhibited a statistically significant correlation with tumour location and macroscopic type according to Bormann's classification. Correlations on statistical borderline were noted between survival time and depth of gastric wall invasion and PCNA PI in the main mass of tumour.

Immunohistochemical evaluation of Ki-67, PCNA and MCM2 proteins proliferation index (PI) in advanced gastric cancer.

Immunohistochemical evaluation of Ki-67, PCNA and MCM2 proteins proliferation index (PI) in advanced gastric cancer.

Ki-67 Expression in Dentigerous Cysts, Unicystic Ameloblastomas, and Ameloblastomas arising from Dental Cysts

This study investigated whether or not an ameloblastoma developing in the wall of a dentigerous cyst is a distinct lesion from the unicystic ameloblastoma. An immunohistochemical evaluation of Ki-67 in dentigerous cysts, unicystic ameloblastomas, and ameloblastomas arising in dentigerous cysts was done. The values of Ki-67 positivity were 3.14 for the dentigerous cyst, between 5.32 and 16.56 for unicystic ameloblastoma, and 11.77 for ameloblastoma arising in a dentigerous cyst. Statistically significant differences were found between the dentigerous cyst and the unicystic ameloblastoma and between the dentigerous cyst and the ameloblastoma arising from a dentigerous cyst. No statistically significant difference was present between unicystic ameloblastoma and ameloblastoma arising from dentigerous cyst. These immunohistochemical data confirm the hypothesis that an ameloblastoma arising from a dentigerous cyst has a similar biological behavior to the unicystic ameloblastoma and should be considered as merely a histologic variant.