Monoclonal Ki-67 Research Articles

Evaluation of Kindlin-1 and Ki-67 immunohistochemical expression in primary cutaneous malignant melanoma: a clinical series

BackgroundThe capacity for prognostic prediction of cutaneous melanoma, one of the most aggressive cancers, is still difficult due to the tumor heterogeneity and lack of reliable tumor markers. The objective of this study is to correlate, through immunohistochemistry, a Ki-67 and Kindlin-1 staining in malignant melanomas with the prognosis of the disease.MethodsA historical cohort study. Immunohistochemistry, using mouse anti-human Kindlin-1 and Ki-67 monoclonal antibodies, was performed using tissue blocks from primary cutaneous melanoma patients treated between 2006 and 2014 at our institution. Information regarding pathological data and outcomes were retrieved from medical records. Statistical analyses were conducted in SPSS version 18.0.ResultsThirty patients were included. The median age was from 50.93 ± 15.31 years old. The expression of Ki-67 was detected in all patients with primary cutaneous melanoma, while Kindlin-1 was negative in two. Kindlin expression was not significantly correlated with Ki-67 expression by Spearman’s rank correlation analysis (P = 0.46), as well as the expression of both markers and the clinical stage (P = 0.34 and 0.18, respectively). Breslow, Clark and mitotic rate were significantly correlated with AJCC stage (P = 0.001).ConclusionOther studies investigating clinical evolution are needed to further test the potential of these markers as possible prognostic markers.

Immunohistochemical Study of Cell Proliferation in Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC), the most common primary liver cancer, with a poor prognosis, is the fifth most common cancer in men and the seventh in women. The study was made on 32 surgically removed liver carcinomas. In order to compare results, we included a group of non-tumor lesions obtained by liver biopsy. Assessment of the proliferative activity of the studied liver lesions was made using immunohistochemical stains with the monoclonal Ki-67 antibody, clone MIB-1 ready-to-use (DAKO Cytomation CA, USA), in the LSAB-HRP work system. To appreciate the proliferation index of Ki-67 (PI Ki-67), we used the semi-automated method of counting the nuclei on digital images. The statistical analysis was performed using SPSS software, Version 20.0 (IBM SPSS Statistics) and Microsoft Office Excel 2007. Mean value of Ki-67 index was 0.4 � 0.2 in normal liver, 3.52 � 0.2 in non-tumor liver lesions and 13.4�7.7 in HCC (p [ 0.001). In chronic hepatitis, PI Ki-67 varied between 2.5 and 5.8 %, with a mean value of 5.2% in portal chronic hepatitis and 5.5% in active chronic hepatitis with cirrhotic evolution. In HCC, the values of Ki-67 index were between 0.7% and 52%, with a mean of PI Ki-67 of 13.43�7.7. 66.6% of HCC associated with hepatitis B virus infection and those developped from a cirrhotic lesion had a high Ki-67 score (p = 0.1). The results we obtained showed: a low Ki-67 score in patients with well-differentiated HCC (G1) (p[0.001), with or without capsule infiltration (p = 0.003); high PI Ki-67 in 33.33% of HCC detected in the right hepatic lobe and those extended bilaterally at the moment of diagnosis (p = 0.142) and a significant relationship between high Ki-67 score and vascular invasion (p [ 0.001), the presence of intrahepatic metastasis being correlated with a high proliferative rate (p [ 0.001). Differences between the proliferation rate of HCC and non-tumor liver lesions (p [ 0.001) show that the uncontrolled division of tumor cells can play an important role in the developpment of HCC.

Abstract 5246: A paradox: Expression of the myelin and lymphocyte protein 2 (MAL2) is down-regulated in human hepatocholangio and renal carcinomas in contrast to its up-regulation in other cancers

Abstract Recent work has found that Myelin and Lymphocyte Protein 2 (MAL2), a protein that functions in polarized protein sorting, is frequently overexpressed in many epithelial-derived human cancers, and has been linked to poor prognosis in patients with colorectal and pancreatic cancers. Yet, its role in tumorigenesis is unknown. Our recent overexpression/knockdown studies in polarized hepatic WIF-B cells, Hep3B hepatocellular carcinoma cells and hepatoma-derived Clone9 cells all indicate that MAL2 is a tumor suppressor. How can MAL2’s function as a tumor suppressor be reconciled with its upregulated expression and link to bad prognosis in human cancers? MAL2 resides on chromosome region 8q24, a region frequently amplified in multiple epithelial-derived human cancers along with c-Myc, a well-known oncogenic driver whose overexpression dysregulates gene expression leading to downstream decreased expression of MAL2 via inactivation of the Miz1 transcription factor. Our prediction is that lower grade lesions from human epithelial-derived cancers display high MAL2 expression that will diminish as cancers progress into higher grade lesions and metastases with a concomitant increase in c-Myc expression. We tested this hypothesis with a cohort, of 23 cholangiocarcinoma (CC), 18 hepatocellular (HCC) and 20 renal cell (RCC) human carcinoma cases whose formalin fixed paraffin embedded tissue sections containing benign and later-stage tumor lesions were immunostained using the Dako Autostainers Link 48 along with Mal2 rabbit polyclonal (ABCAM), Miz-1 monoclonal ZBTB17(NOVUS), c-Myc monoclonal Y69 (ABCAM) and Ki-67 monoclonal Mib-1(Dako) antibodies. Nuclear and/or cytoplasmic immunoreactivity was scored based on the intensity and percentage of positive cells in both the benign epithelium and adjacent tumor component in each case. In all three carcinoma types, both MAL2 and Miz-1 expression was high in the benign tissue. In the tumor component MAL2 and Miz-1 expression was respectively lost in 15/23(65%) and 22/23(96%) CC, 13/18(72%) and 17/18(94%) HCC, 13/20(65%) and 16/20(80%) RCC cases. In contrast only 2/20 (10%) and 1/20(5%) RCC cases showed an upregulated expression of MAL2 and Miz-1 respectively compared to the benign component, with all three remaining carcinoma cases showing no up-or-down regulation in protein expression. However, no enhanced c-Myc expression was detected in the tumor lesions despite the higher proliferative index as indicated by Ki-67 labeling. These results indicate a concordant down-regulation of both MAL2 and Miz-1 in CC, HCC, and RCC supporting a tumor suppressor role in these cancers. We are currently extending these studies to other epithelial-derived cancer tissue types and also are more closely examining MAL2, Miz1 and c-Myc expression across graded lesions to better test our hypothesis. Note: This abstract was not presented at the meeting. Citation Format: Alfonso Lopez-Coral, Joeffrey J. Chahine, Bhaskar V. Kallakury, Pamela L. Tuma. A paradox: Expression of the myelin and lymphocyte protein 2 (MAL2) is down-regulated in human hepatocholangio and renal carcinomas in contrast to its up-regulation in other cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5246.

Prognostic Factors for Luminal B-like Breast Cancer.

This study aimed to examine the prognostic factors of luminal B-like breast cancer. Clinical data of 695 luminal B-like breast cancer patients who had been treated in our hospital during the period of past 4.5 years were collected and analyzed. Estrogen receptor (ER), progesterone receptor (PgR), antigen identified by monoclonal antibody Ki-67 (Ki67) were immunohistochemically detected. Different cutoffs of ER, PgR, and Ki67 were evaluated. Pearson χ2 test was performed to compare categorical parameters. Univariate and multivariate models were used to evaluate predictors of disease free survival (DFS). The results showed that patients who were younger, and had larger tumors, and more positive lymph nodes were more likely to receive neo-adjuvent chemotherapy (NAC). Patients with ER-positive tumors having <10% positive cells received more anthracycline- and taxane-based chemotherapy and less endocrine therapy than those with ER-positive tumors having ≥10% positive cells (P=0.004 and P=0.007, respectively); however, patients with ER-positive tumors having <10% positive cells experienced more recurrence (P<0.001). PgR expression levels were not associated with therapeutic schedule and DFS. Patients with tumor tissue Ki67 score ≥30% received more anthracycline- and taxane-based chemotherapy and had worse DFS than those with tumor tissue Ki67 score <30%. Univariate and multivariate analysis showed that clinical T stage, lymph nodes, ER, Ki67, and HER2 status were independent prognostic factors. In conclusion, ER-positive rate <10% and Ki67 score ≥30%, similar to higher clinical T stage, more metastatic lymph nodes, and HER2 positive status, may indicate a worse prognosis for luminal B-like breast cancer patients. Multi-center prospective trials with larger sample sizes are necessary for the continued perfection of our work.

Correlation between TopIIα and ki-67 expression in tumors of luminal, primary operable breast cancer without HER2/neu overexpression in postmenopausal women as a prognostic factor.

e12065 Background: Assessment of proliferative activity is important in terms of tumor prognosis. Topoisomerase IIα (TopIIα) and ki-67 are common markers of breast cancer (BC) cell proliferation. The purpose of the study was to assess the correlation between TopIIα and ki-67 expression as a prognostic factor in luminal, primary operable breast cancer without Her 2 neu overexpression in postmenopausal women. Methods: 60 postmenopausal patients with BC (invasive ductal carcinoma) were equally divided into 3 groups: I –luminal A BC, surgical treatment with subsequent radiotherapy and hormonal therapy; II – luminal B BC, surgical treatment with subsequent radiotherapy and hormonal therapy; III – luminal B BC, surgical treatment with subsequent radiotherapy, polychemotherapy and hormonal therapy. Mean age of patients: I – 76.16±3.14 years, II – 65.0±3.91, III – 74.0±2.54 years. Immunohistochemical study was performed with mouse monoclonal ki-67 (Thermo scientific) and TopIIα (Santa Cruz Biotechnology) antibodies using the Reveal Polyvalent HRP-DAB Detection System. Results were analyzed using the STATISTICA 10.0 program (StatSoftInc., USA). Results: The average expression levels of the ki-67 marker in the groups were: I - 12.3±1.4%, II - 15.1±3.1%, III - 37.0±7.1%; TopIIα expression – 3.5±1.6%, 16.0±5.9%, 21.7±6.4%, respectively. The statistically significant Spearman correlation between TopIIα and ki-67 was observed both as general in groups (r = 0.787 at p &lt; 0.001) and in groups with luminal B subtype (groups II and III). Strong positive correlation between TopIIα and ki-67 was registered in groups II (r = 0.941 at p &lt; 0.01) and III (r = 0.864 at p &lt; 0.001) with luminal B subtype. Weak correlation was observed in group I with luminal A subtype (r = 0.477 at p≥0.05). Conclusions: The study revealed a correlative increase in the level of TopIIα with simultaneous overexpression of the ki-67 protein in the studied groups. This indicator can be considered predictive for breast cancer.

C-ERB-2, P53, Ki67 Proteins and Receptors of Estrogen and Progesterone on the Prognosis of Epithelial Ovarian Cancer

Ovarian cancer is the seventh most common cancer diagnosed in women worldwide. To date, many studies inepithelial ovarian cancer (EOC) have reported on the association HER-2/neu, p53 proteins and steroid hormones and their respective receptors with prognosis and/or the carcinogenesis process, but no definitive conclusion has been reached.Objectives: To assess the proteins c-erbB-2, p53, Ki67 and receptors of estrogen (ER) and progesterone (PR) of EOC, with regard to clinical stage findings and its effect on survival.Methods: 125 patients with a diagnosis of EOC treated by primary surgery and chemotherapy have participated. A surgical stage was noted and analyzed the correlation with c-erbB-2, p53, Ki67, ER and PR. Immunohistochemical analysis, using the anti-c-erbB-2, p53, Ki67 monoclonal antibodies, the antibody cod PR clone PgR and code ER-6-F11 Anti human estrogen. The c-erbB-2 study was complemented by genetic amplification and was reported univariate and multivariate analysis.Results: Age 55.7 ± 16; 50.2% with residual disease (&lt; 2 cm); initial (54.6%) and advanced (45.4%) stage. Univariate analysis showed positive staining for c-erbB-2, p-53, Ki67, PR and ER. The patients with negative receptors had a significantly shortened survival time (p = 0.01) than patients with positive receptors. Multivariable analysis revealed only clinical FIGO stage as an independent prognostic of overall survival (p = 0.002). Other variables like c-erbB-2, p53, Ki67, and ER were not significantly related to survival.Conclusions: We concluded that patients with negative PR had a significantly shortened survival time than patients with positive receptors. The overexpression of markers c-erbB-2, p53, Ki67, and ER, were not significantly related to survival in EOC. Only the FIGO stage was achieved to be an independent predictor of overall survival. They should be evaluated together with the patient’s clinical status and other prognostic factors.

Immunohistochemical study of Ki-67, PHH3, and CK15 protein expression in oral epithelial malignancy

To investigate the expression of Ki-67, phosphohistone H-3 (PHH3), and cytokeratin 15 (CK15) proteins in the cells of the oral mucosa (OM) according to the degree of its malignant transformation. OM biopsy specimens from 69 patients diagnosed with focal epithelial hyperplasia, intraepithelial squamous cell neoplasia, cancer in situ, and squamous cell carcinoma were examined. Tissue antigens were determined using mouse Ki-67 monoclonal antibodies, rabbit PHH3 polyclonal antibodies, and mouse CK15 monoclonal antibodies. There was an increase in epithelial proliferative and mitotic activities in squamous cell carcinoma and a sharp decrease in the expression of CK15 in the cytoplasm in cancer in situ and squamous cell carcinoma of the OM. The protein CK15 can be used for the differential diagnosis between high-grade dysplasia and OM epithelial malignancy at the stage of carcinoma in situ and squamous cell carcinoma.

Ki-67 Prognostic Value in Different Histological Grades of Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma

Introduction:Abnormal cell proliferation appears to be a possible predictor of tumorigenesis, Ki-67 protein expression is closely related to the cell proliferation and could be used as a biomarker for the growth in the most of human tumors. The aim of the study: Investigating of Ki-67 expression in the pathological grades of oral epithelial dysplasia and oral squamous cell carcinomas.Materials and Methods:The sample consisted of 30 formalin-fixed, paraffin-embedded specimens of oral epithelial dysplasia (OED), 30 other of oral squamous cell carcinomas (OSCC), and 10 normal oral epithelium (NOE) were conventionally stained with hematoxylin and eosin and immunohistochemically stained with Ki-67 monoclonal antibody.Results:Expression of Ki-67 was restricted to the basal layers in the normal oral epithelium whereas Ki-67 positive cells in oral epithelial dysplasia (OED) were located in the basal, suprabasal and spinous layers, Ki-67 expression was increased in high-risk cases. Ki-67 positive cells in well-differentiated (OSCC) were located mainly in the periphery of the tumor nests, in moderately-differentiated (OSCC) were located in both peripheral and part of a center of the tumor nests whereas it was diffused in most of the Poorly-differentiated (OSCC). Statistical analysis indicated a significant difference between the expression in (OED) and (NOE), (OSCC) and (NOE), and no differences between (OED) and (OSCC).Conclusion:This study has concluded that Ki-67 antigen could be used as a marker for the histological grading of OED and OSCC, Expression of Ki 67 increased according to the severity of oral epithelial dysplasia.

Assessment of markers expressed in human hair follicles according to different skin regions.

Body region-dependent hair follicle (HF) characteristics are concerned with follicular size and distribution, and have been demonstrated to have characteristics for each region of the body. The aim of the present study was to investigate the expression patterns of the markers called cytokeratin 15 (K15), cytokeratin 6 (K6) and monoclonal antibody Ki-67, and also apoptosis in HFs, which can be observed in different parts of the human body. In this study, healthy human HFs were taken by biopsy from 5 various donor sites of the human body: the scalp, the leg, the abdomen, the back and waist. HF-containing skin specimens taken using cryosection were stained with hematoxylin & eosin (H&E) and K15, K6, Ki-67 and terminal deoxynucleotidyl transferase-mediated digoxigenin-dNTP nick end-labelling (TUNEL) immunofluorescence staining protocol was performed. Different skin regions from the human body were examined histologically. While the HFs of scalp tissue showed anatomically obvious hair layers, some hair sections from other regions, like the leg, the abdomen, back and waist, were not as distinct as in the scalp region. According to our findings, K15 expression was highest in the scalp. In addition, the immunoreactivity (IR) intensity of K15 was significantly decreased in the HFs on the waist and abdominal regions, compared to the scalp and back regions (p < 0.001). However, the IR intensity of K6 in the scalp region was statistically significantly higher than the IR intensity of K6 in the abdomen region (p < 0.05). Moreover, we showed intraepithelial apoptosis and proliferation of keratinocytes in the bulge of HF. In the study, Ki-67-positive and TUNEL-positive cell numbers were not statistically significant (p > 0.05). Our findings are important for further investigation of molecular aspects of the human hair follicle stem cells compartments in health and disease, which might be a promising model for comparative studies with different human diseases.

Vasoactive intestinal peptide induces proliferation of human hepatocytes.

Proliferation of hepatocytes in vitro can be stimulated by growth factors such as epidermal growth factor (EGF), but the role of vasoactive intestinal peptide (VIP) remains unclear. We have investigated the effect of VIP on maintenance and proliferation of human hepatocytes. Human hepatocytes were isolated from liver specimens obtained from patients undergoing liver surgery. Treatment with VIP or EGF was started 24h after plating and continued for 3 or 5d. DNA replication was investigated by Bromodeoxyuridine (BrdU) incorporation and cell viability detected by MTT assay. Cell lysate was analysed by western blotting and RT-PCR. Urea and albumin secretion into the culture supernatants were measured. VIP increased DNA replication in hepatocytes in a dose-dependant manner, with a peak response at day 3 of treatment. VIP treatment was associated with an increase in mRNA expression of antigen identified by monoclonal antibody Ki-67 (MKI-67) and Histone Cluster 3 (H3) genes. Western blotting analysis showed that VIP can induce a PKA/B-Raf dependant phosphorylation of extracellular signal-regulated kinases (ERK). Although EGF can maintain hepatocyte functions up to day 5, no marked efffect was found with VIP. VIP induces proliferation of human hepatocytes with little or no effect on hepatocyte differentiation. Further investigation of the role of VIP is required to determine if it may ultimately support therapeutic approaches of liver disease.

Analysis of membranous Ki-67 staining in breast cancer and surrounding breast epithelium

Membranous Ki-67 staining with the MIB-1 antibody has been described in hyalinising trabecular adenomas of the thyroid and sclerosing haemangiomas of the lung. Its relatively rare occurrence in breast tumours has also been documented. The aim of the present study was to assess the rate of any membranous MIB-1 staining in breast specimens. The staining was performed at room temperature with 1:100 dilution of the antibody. One hundred four core needle biopsies and 41 operative specimens were analysed. Membranous staining was noted in 36/144 invasive carcinomas, 20/42 in situ carcinomas and 46/99 cases of peritumoural benign/normal breast epithelium. Most often, it presented as focal and partial polarised luminal membranous staining although complete circumferential staining also occurred, and membranous labelling was sometimes accompanied by cytoplasmic staining, too. In a few cases tested, greater dilution of the primary antibody did not abolish the membranous staining, which was absent with the SP6 monoclonal Ki-67 antibody. The membranous staining of invasive tumours showed no association with histological grade, lumen formation, oestrogen or progesterone receptor status or the Ki-67 nuclear labelling. In contrast, it was associated with a HER2-positive status, although it occurred in all molecular subtypes approached by immunohistochemistry. The background of this membranous staining remains elusive. It is unlikely to represent an artefact. At least partial sharing of an epitope of the nuclear Ki-67 protein with an unidentified membranous protein and some functional differences between membranous staining producing tumours and tumours lacking this pattern of staining may both contribute to some extent.

Vitexin reverses the autophagy dysfunction to attenuate MCAO-induced cerebral ischemic stroke via mTOR/Ulk1 pathway

Stroke, as a kind of acute cerebrovascular diseases, has greatly influenced the patients’ quality of life and left a huge public health burden. Vitexin is a flavone C-glycoside (apigenin-8-C-β-D-glucopyranoside) present in several medicinal and other plants. This study aims to explore the role of vitexin in middle cerebral artery occlusion (MCAO)-induced cerebral ischemic stroke. The results showed that the MCAO-induced brain infarction was obviously decreased by vitexin. And the abnormal protein levels of Caspase-3, Bcl-2-associated X protein (Bax), antigen identified by monoclonal antibody (Ki-67) and B cell lymphoma 2 (Bcl-2) in MCAO model rats were reversed by vitexin. Further research indicated that vitexin alleviated MCAO-induced oxidative injury by reducing the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA) and nitric Oxide (NO). In addition, vitexin attenuated the secretion of pro-inflammatory cytokine (interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α)) and increased anti-inflammatory cytokine (IL-10) production to ameliorate MCAO-induced inflammation. What’s more, vitexin repressed the MCAO-induced autophagy through mechanistic target of rapamycin (mTOR)/Ulk1 pathway. Specifically, the MCAO-induced decreased expression of mTOR, peroxisome proliferator-activated receptor γ (PPARγ) and p62 were inhibited by vitexin. At the same time, MCAO-induced increased expression of Ulk1, Beclin1 and rate of LC3Ⅱ/LC3Ⅰ also were repressed by vitexin. But the inhibition of vitexin on the MCAO-induced oxidative injury, apoptosis and inflammation were reversed by rapamycin. These results implied that vitexin suppressed the autophagy dysfunction to attenuate MCAO-induced cerebral ischemic stroke via mTOR/Ulk1 pathway.

Abstract A15: The relationship between cervical cancer associated genes and human papilloma viruses in rural Africans in Zaria

Abstract Cervical cancer is a major public health problem and one of the leading gynecological cancers in developing nations. Reports from Nigeria, the largest black nation in the world by population, show that the national incidence of cervical cancer is 250/100,000. We sought to evaluate the relationship between cervical cancer associated genes and human papilloma viruses from rural Africans in Zaria, Nigeria using immunohistochemistry and molecular phylogenetics. Selected cervical cancer associated genes sequences which include Ki-67 (NM_002417.4), p53 (AB082923.1), EGFR (NM_005228.3), MYC (M25803.1), KRAS (EF471957.1), TGF-B (AF254794.1) and RB (AH002963.2) genes and HPV 16 (KJ869425.1 E6 and E7 variants), HPV 18 (KJ869426.1), HPV 45 (S40261.1), HPV 6 (A25369.1), HPV 11 (J04351.1), HPV 31 (KJ869427.1) and HPV 33 (KJ869428.1) sequences were retrieved from the gene bank data base. The sequences were aligned and bootstrap at 2000 bp to create a molecular phylogenetic tree. Haematoxylin and Eosin together with immunohistochemistry were performed on fifty formalin-fixed paraffin-embedded (FFPE) tissue blocks using Anti- Ki-67, p53, EGFR, MYC, KRAS, TGF-B and RB monoclonal antibodies. Result from the phylogenetic analysis reveals trichotomized tree showing apparent significant evolutionary relationship between cladogram of HPV16 and MYC; HPV18, HPV6 and KRAS genes; EGFR gene and HPV45. Immunohistochemistry results show multivariate expression of Ki-67, p53, EGFR, MYC, KRAS, TGF-B and RB of proteins from the cervical cancer tissue blocks used. Combination of Ki-67 biomarker and other cervical cancer associated genes together with HPV subtypes-genotyping can serve as useful instruments in proper management of the disease. Citation Format: John Idoko, Mohammed Faruk, Jigo Dangude Yaro, James Oluwa Enemari, Peter Akpulu. The relationship between cervical cancer associated genes and human papilloma viruses in rural Africans in Zaria [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr A15.

Morphological characteristics of uterine fibroids among women of reproductive age

The article deals with modern views on etiopathogenesis of uterine fibroids. Increased knowledge of the markers of proliferation and their study will help to predict the course of the disease and thus to develop the optimum tactics of the further treatment of the early stages. Much attention is paid to the study of markers that characterize apoptosis. By immunohistochemistry in tissue fibroids 15 patients studied protein expression Ki-67 and Bcl-2. Analyzed values of the correlation expression of the proteins according to the clinical and morphological features of the tumor. It showed a reduction in the proliferation marker Ki-67, and increased expression of apoptotic Bcl-2 inhibitor.Background. Presented by changes in the nature of proliferation and apoptotic activity in myoma nodes among women of reproductive age who underwent conservative myomectomy.The aim of the research was to study the performance of protein antibody Ki-67 proliferation and apoptosis inhibitor of Bcl-2 in women with uterine myoma.Material and methods. Surveyed 15 women with uterine fibroids who underwent conservative myomectomy. Inclusion criteria were: the reproductive age, uterine fibroids the size of which does not exceed the 12-week pregnancy. Exclusion criteria were the size of fibroids greater than 12 weeks of pregnancy, acute inflammatory diseases of the pelvic organs, severe somatic diseases, cancer of the reproductive organs. It was carried out histological and immunohistochemical study of remote myoma node. We examined by immunohistochemistry proliferation protein monoclonal antibody Ki-67 (Clone MIB-1, 1 : 150, Dako), inhibitor of apoptosis Bcl-2 (NCL-bcl-2-486). (Novocastra). Evaluation of the expression of markers studied conducted qualitative and quantitative methods in computer image analysis system “Morphology 5.0” (Video Test, Russia). Statistical processing of the results was performed using statistical analysis packages (STATGRAPHICS v. 6.0).Results. Age of women surveyed averaged 33,4 ± 4,1 years. Histological study of benign tumors of the uterus body showed the presence of degenerative changes, necrosis and inflammatory infiltration in every fourth case of the material.Conclusions. Immunogistohimichesoe study of uterine leiomyomas body revealed a significant increase in the expression of apoptosis inhibitor of Bcl-2 and reducing the proliferation marker Ki-receptor expression in all 67 tested samples, which is consistent with a histological study of the absence of mitotic activity in leiomyomas.

Differential Expression of Glypican-3 and Insulin–Like Growth Factor-II mRNAs and Alpha-Fetoprotein and Ki-67 Markers in HCV Related Hepatocellular Carcinomas In Egyptian Patients

Background:Increasing evidence indicates that in hepatocellular carcinomas (HCCs) abnormal gene expression, for example of glypican-3 (GPC-3) and insulin-like growth factor-II (IGF-II), are associated with the occurrence and progression of HCC. The objective of this study was to evaluate the differential expression of GPC-3 and IGF-II mRNAs in HCC tissues with a background of chronic hepatitis C virus (HCV) genotype 4 cirrhosis, in relation to Ki-67 and alpha-feto protein (AFP) tissue markers.Methods:One hundred and five patients with HCCs who had undergone hepatectomy, were included, after obtaining informed consent. Total RNA was extracted from malignant and corresponding peri-malignant liver tissues, and GPC-3 and IGF-II mRNAs in addition to beta-actin mRNA as an internal control, were evaluated in all samples by reverse transcriptase-polymerase chain reactions (RT-PCR). Routine histopathological diagnosis as well as immunohistochemical (IHC) staining using monoclonal antibodies for Ki-67 and AFP were also performed.Result:Expression of GPC-3 mRNA was positive in all HCC malignant tissue, with overexpression in 86/105 (81.9%); in respect to the grade of the tumor (1-3 grades), while in peri-malignant tissue it was over expressed only in 20/105 (19%). The IGF-II mRNA was over expressed in only 10/105 (9.5%) malignant and peri-malignant samples. AFP was expressed in 33.3% of malignant samples but absent in peri-malignant tissues. Ki-67 expression was significantly increased in malignant compared to peri-malignant tissue.Conclusion:GPC-3 and IGF II mRNAs may be good molecular markers for HCC, especially with a background of cirrhosis due to chronic HCV infection. Significant correlations were noted with the pattern of AFP and Ki-67 expression.

Comparison of Core Needle Biopsy and Surgical Specimens in Determining Intrinsic Biological Subtypes of Breast Cancer with Immunohistochemistry.

PurposeWe evaluated the concordance between core needle biopsy (CNB) and surgical specimens on examining intrinsic biological subtypes and receptor status, and determined the accuracy of CNB as a basic diagnostic method.MethodsWe analyzed breast cancer patients with paired CNB and surgical specimen samples during 2014. We used monoclonal antibodies for nuclear staining, and estrogen receptor (ER) and progesterone receptor (PR) status evaluation. A positive test was defined as staining greater than or equal to 1% of tumor cells. Human epidermal growth factor receptor 2 (HER2) was graded by immunohistochemistry and scored as 0 to 3+ according to the recommendations of the American Society of Clinical Oncology/College of American Pathologists. Ki-67 immunostaining was performed using the monoclonal antibody Ki-67, and the results were divided at 10% intervals. The cutoff value for high Ki-67 was defined as 20%. Concordance analysis of ER, PR, HER2, Ki-67, and five intrinsic biological subtypes was performed on CNB and surgical specimens. Statistical analysis for concordance was calculated using κ-tests.ResultsWe found very good agreement for ER and PR with a concordance of 96.7% for ER (κ=0.903), and 94.3% for PR (κ=0.870). HER2 and Ki-67 showed concordance rates of 84.8% (κ=0.684) and 83.5% (κ=0.647), respectively, which were interpreted as good agreement. Five subgroups analysis showed 85.8% agreement and κ-value of 0.786, also indicating good agreement.ConclusionCNB showed high diagnostic accuracy compared with surgical specimens, and good agreement for ER, PR, HER2, and Ki-67. Our findings reaffirmed the recommendation of CNB as an initial procedure for breast cancer diagnosis, and the assessment of receptor status and intrinsic biological subtypes to determine further treatment plans.

The immunomorphological features of cervical intraepithelial neoplasia associated with HPV infection depending on the type of infertility

Aim . Objective was to establish the features of markers expression of receptivity, proliferation, apoptosis and angiogenesis of cervix with CIN associated with HPV infection in various forms of female infertility. Materials and methods . Cervical biopsy material of 157 women with CIN associated with HPV infection in infertility was investigated. Depending on the degree of CIN all cases are divided into three groups: 1 st group – 62 women with mild CIN (CIN-I), 2 nd group – 53 women with moderate CIN (CIN-II), 3 rd group – 42 patients with severe CIN (CIN-III). The control group consisted of 15 cervical samples from infertile women without cervical pathology. For immunohistochemical (IHC) studies the primary monoclonal antibodies Ki-67 (Clone MIB-1, DakoCytomation) r63 (clone 4A4, DakoCytomation), VEGF (clone VG1, DakoCytomation), p16ink4a (kit for histological preparations, number K5334), estrogen (ER) (clone 6F11, Novocastra) and progesterone receptors (PR) (clone 1A6, Novocastra) were used. Results. As a result of studies it was found that dysplastic cervical processes in terms of human papilloma virus infection are characterized by severe structural changes in the form of cell renewal violation, redistribution of hormonal reception and expression of vascular endothelial growth factor by stratified squamous epithelium especially in the tubular, hormonal and combined infertility. Conclusions . It was established that for infertility the cervical intraepithelial neoplasia (CIN) associated with HPV infection (PVI) is characterized by different individual potential for the development of cervical carcinoma. For early detection, verification of the severity degree and prognosis of CIN, especially during the tubular, hormonal and combined forms of infertility it is advisable to include the immunomorphological study of tissue of the cervix using monoclonal antibodies (Ki-67, r63, pl6ink4a, ER, PR, VEGF) in the algorithm of complex inspection.

The use of molecular markers (Ki67 &amp; p53) in premalignant and malignant cervical neoplasms

Introduction: Cervical cancers are the second most frequent type of cancer in females. It develops from premalignant to invasive stages in a multistep process of carcinogenesis. The differentiation between cervical intraepithelial neoplasia III (CIN III) and early squamous cell carcinoma (SCC) of the cervix could be difficult to diagnose in certain situations unless we see the invasion. The objective of the study was to determine the expression of p53 & Ki67 in CIN and cervical cancers and its correlation with grading and differentiation of cervical cancers with special emphasis on whether the use of Ki67 & p53 markers could help in differentiating CIN III from early SCC. Material and Methods: A prospective study on 90 cases of cervical biopsies, including- cases of : chronic cervicitis (40), CIN III (19) and invasive carcinoma (31). All cases were evaluated by immunohistochemistry using Ki67 & p53 monoclonal antibodies. Results: Both Ki67 & p53 showed higher expression in CIN III and invasive cervical carcinoma than chronic cervicitis with statistically significant difference p<0.01 and p<0.001 respectively. High significant difference of p53 & Ki67 expression was found between patients with CIN III and invasive cervical carcinomas. There was a clear trend for increasing number of cases with positive Ki67 & p53 index with increasing grades of cervical cancer. Conclusion: Ki67 & p53 showed increased expression from CIN III to SCC and it may be helpful adjunct in differentiating these two lesions. These markers could be used as biomarkers in the evaluation of the proliferative activity and progressive potential of dysplastic and neoplastic changes.

Interaction between Esophageal Squamous Cell Carcinoma and Adipose Tissue in Vitro

Esophageal squamous cell carcinoma (ESCC) develops within the squamous epithelial layer and invades the submucosa to the subadventitia that has adipose tissue (AT). AT seems critical to ESCC progression, but the underlying mechanism is unknown. We aimed to address the association between ESCC and AT invitro. ESCC cells were cultured on rat or human subcutaneous AT-embedded or -non-embedded collagen gel. AT promoted the growth of ESCC cells and inhibited their apoptosis. AT promoted the expression of the squamous differentiation marker involucrin in ESCC cells. AT accelerated the expression of invasion-related factors in poorly differentiated ESCC cells only. AT promoted the expression of phosphorylated-insulin-like growth factor-1 receptor in ESCC cells, whereas it inhibited that of the human epidermal growth factor receptor 2. Insulin-like growth factor-1, but not leptin, adiponectin, or resistin, promoted and inhibited the growth and apoptosis of ESCC cells, respectively. In turn, ESCC cells decreased the production of these adipokines in AT and the number of preadipocytes and mesenchymal stem cell-like cells, which developed from AT. These results suggest that i) AT may influence the progression of ESCC with increased growth or invasion and decreased apoptosis through insulin-like growth factor-1/insulin-like growth factor-1 receptor signaling, ii) AT may affect human epidermal growth factor receptor 2-targeted therapy; and iii) the cancer cells may affect adipokine production in AT.

Study of Ki67 and CD10 expression as predictive factors of recurrence of ameloblastoma

IntroductionAmeloblastoma is a rare, benign, purely epithelial odontogenic tumour, characterized by a high potential for local invasion and recurrence. ObjectiveTo study the epidemiological and histological characteristics of ameloblastoma. To study Ki67 and CD10 immunostaining in ameloblastoma and to investigate a possible correlation between these two markers and recurrence of this tumour. MethodsAn immunohistochemical study using Ki67 and CD10 monoclonal antibodies was performed on 37 paraffin blocks obtained from the Charles-Nicolle hospital pathology department in Tunis over a 9-year period (2004–2012). Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) software version 15.1. ResultsThis series of 37 cases comprised 21 males and 16 females (sex ratio: 1.3) with a mean age of 39 years (range: 7 to 70 years), corresponding to 36 cases of intraosseous ameloblastoma and one case of gingival ameloblastoma. Thirty-two cases were polycystic and 5 cases were unicystic. Eighteen cases of local recurrence were observed. No correlation was demonstrated between recurrence and the various clinical and histological parameters and treatment modalities. However, a significant correlation was demonstrated between recurrence and Ki67 and CD10 expression (P=0.000 and 0.002, respectively). ConclusionThe Ki67 proliferation index and stromal CD10 expression can be considered to be predictive factors of ameloblastoma recurrence.