Localization Of Kaiso Research Articles

Kaiso Expression in Triple Negative Breast Cancer in a Tertiary Hospital in Ghana.

Breast cancer has produced more lost disability-adjusted life years (DALYs) than any other type of cancer. The prevalence of the disease, especially triple negative breast cancer (TNBC) in Africa is on the rise, with poor survival rates. With the great advancements in treatments of breast cancers, that of TNBC is still a challenge due to its narrowed treatment options and poor disease prognosis. This research seeks to explore the expression of kaiso in Ghanaian breast cancer and how they may modulate clinicopathological features, and disease prognosis. A cross-sectional retrospective study was conducted on formalin-fixed paraffin-embedded (FFPE) breast cancer tissues retrieved from the archives of the pathology unit of Komfo Anokye Teaching Hospital (KATH). Immunohistochemistry assessment was performed on haematoxylin and eosin-stained slides selected for tissue microarray construction. Data were analysed using SPSS version 28 and Microsoft excel 2013. 55.3% of the cases tested negative to progesterone receptor (PR), oestrogen receptor (ER), and human epidermal growth receptor 2 (HER2). There were significant associations between menopausal status and molecular subtype (p=0.010), Kaiso expression and histological diagnoses (<0.001) and Kaiso against lymphovascular invasion (0.050). However, there were no significant associations between Kaiso localization and the clinicopathological features although 63.9% of the expression was seen in the nucleus. The study indicates that Kaiso is highly expressed in Ghanaian TNBC and likely associated with worse outcomes in aggressive tumour types.

Association of Kaiso and partner proteins in oral squamous cell carcinoma

Objectives:1. Identification of protein expression and subcellular localization of E-cadherin (E-cad), p120 catenin (P120ctn), and Kaiso in oral cancer (OC). 2. To study the protein expression of cyclin D1 and c-Myc (Kaiso targets) and determine their relationship with the expression and localization of Kaiso. MethodsHistological grading was performed in accordance with Broder's criteria. Expression and localization data for E-cad, p120ctn, Kaiso, cyclin D1, and c-Myc were acquired using immunohistochemistry. Data were analyzed using SPSS version 21. The chi-square test was used to measure the statistical significance of associations, with p < 0.05 as statistically significant. ResultsOf 47 OC cases, 36% showed low E-cad expression and 34% showed low p120ctn. Low Kaiso expression was recognized in 78% of tumor specimens. Aberrant cytoplasmic localization of p120ctn was seen in 80.8% cases. Cytoplasmic Kaiso localization was appreciated in 87% of tumor tissues, whereas 29.7% lacked any nuclear Kaiso. Kaiso expression was significantly associated with the expression of cyclin D1 but not with c-Myc. ConclusionThe present study identified a change in the localization of Kaiso in OC. The significance of this in relation to OC and tumor prognosis needs to be investigated with further studies using larger sample sizes and more sensitive molecular tools.

Abstract 1680: Targeting subcellular localization of Kaiso to limit dissemination through MAPK signaling

Abstract Breast cancer is the second most commonly diagnosed cancer among American women. The American Cancer Society has estimated over 230,000 new cases of breast cancer, and over 40,000 deaths for the year 2015. Unfortunately, metastasis is the major contributor for the significant number of these deaths. The Epithelial-Mesenchymal Transition (EMT) is promoted by the Master Regulator known as Kaiso, a transcriptional repressor that is a member of the BTB/POZ family of zinc finger transcription factors. We have previously demonstrated that Epidermal Growth Factor Receptor (EGFR) signaling shuttles Kaiso into the nucleus, although, its mechanism is still not understood. EGFR signaling activates multiple downstream signaling cascades including the MAP Kinase cascade, which has a significant influence on proliferation, cell adhesion, gene expression, and cell survival. Mesenchymal cells demonstrate high levels of MAPK activity and disruption or inhibition of the cascade has proven to be a method for decreasing tumor growth. Therefore we hypothesize that subcellular localization and expression of Kaiso is modulated by EGFR induced MAPK activity. Using the TCGA Breast cancer dataset, we observed that Kaiso was altered in about 11% of patients, compared to 15% of E-cad expression and 13% of MAPK3 of 959 patients/cases of Breast cancers. MAPK3 (eRK1); MAPK6, MAPK7, MAPK9 AND MAPK14 (p38) seems to have trends in mutual exclusivity or co-occurrence with Kaiso (ZBTB33). To determine if MAPK3 is responsible for increased Kaiso expression, MCF-7 cells, which express low levels of Kaiso in the cytoplasm, were treated with 10ng/ml of EGF in the presence or absence of MAPK inhibitor, 20μM PD98093. EGF stimulation caused robust increase in Kaiso protein expression, while PD98093 inhibited EGF induced expression. Similarly, in Kaiso overexpressing, MDA-MB-231, blocking EGFR kinase activity with 500nM PD153035 resulted in an opposite effect as EGF stimulation in MCF-7 cells. Furthermore, we observed a significant decrease in p-ERK expression, re-expression of E-cadherin, and low EGFR expression levels in sh-Kaiso MDA-MB-231 cells compared to sh-Scr cells. The decreased p-ERK expression in sh-Kaiso MDA-MB-231 cells was also associated with decreased expression of mesenchymal markers ZEB1, ZEB2, Twist and SNAIL at both the mRNA and protein levels compared to the scrambled control. Similarly, we observed significant decrease in the somatic cell reprogramming factors Klf4, Oct4, Nanog expression in sh-Kaiso MDA-MB-231 as well, which coincide with a reversal of EMT. To our knowledge, this is the first study to investigate the molecular mechanism and/or functional consequences of increased Kaiso expression in breast cancer, and highlights the multiple roles that Kaiso plays in promoting aggressive disease. Citation Format: Shweta Tripathi, Balasubramanyam Karanam, Bo Ma, Alan Wells, Clayton Yates. Targeting subcellular localization of Kaiso to limit dissemination through MAPK signaling. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1680.

Abstract LB-147: Characterization of a Kaiso overexpressing transgenic mouse model

Abstract Introduction: The WNT signaling pathway plays critical roles during embryogenesis and maintaining homeostasis of adult tissues. However constitutive WNT signaling leads to tumorigenesis. Upon activation of the WNT pathway, the main downstream effector of the WNT pathway, ß-catenin, stabilizes and translocates to the nucleus where it activates transcription of WNT target genes. Recently, our lab along with others identified the novel transcription factor Kaiso as a negative regulator of WNT target genes in mammalian cell lines and Xenopus models. However a study using a Kaiso depleted mouse model implicated Kaiso as a positive regulator of WNT signaling. Kaiso is a member of the BTB/POZ (broad-complex, tramtrack and bric-à-brac/poxvirus and zinc finger) protein family. The goal of this study is to elucidate the role of Kaiso in WNT signaling using the ApcMin/+ mouse model of intestinal cancer. Methodology: To assess the role of Kaiso in colon cancer in vivo, Kaiso overexpressing transgenic mice were created (KaisoTg). Targeted overexpression of Kaiso in KaisoTg mice was obtained by pronuclear injection of Kaiso cDNA under the control of the intestine-specific villin promoter. Reverse transcription PCR (RT-PCR) and Western Blot analyses were used to determine tissue specific Kaiso overexpression in KaisoTg mice. Immunohistochemisty (IHC) was performed on mouse intestinal tissues to examine the subcellular localization of Kaiso and assess developmental defects. KaisoTg mice were then mated with APCMin/+ mice models of intestinal cancer that develop polyps in the small intestine due to constitutively active -catenin. KaisoTg-APCMin/+ mice were examined for gross morphological phenotypes and IHC was performed on mouse intestinal tissues for Kaiso and other WNT target genes (e.g. cyclin D1) to elucidate the effects of Kaiso on the WNT pathway. Results: Western Blot analysis revealed high, moderate and low levels of Kaiso overexpression in three founder lines of the KaisoTg mice. RT-PCR analysis showed Kaiso overexpression limited to villin positive tissues including kidney, small and large intestines. IHC analysis of KaisoTg tissues showed no gross morphological differences compared to non-transgenic mice. However KaisoTg-APCMin/+ displayed decreased body size and weight, and had decreased life span when compared to the parentals (i.e. KaisoTg and APCmin/+). IHC analysis of KaisoTg-APCMin/+ mouse intestines revealed increased polyp numbers with Kaiso staining concentrated near the basal surface of the polyps. These data support Kaiso's potential role as an oncogene in mouse intestinal tissues. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-147.

Abstract LB-271: The biological role of the transcription factor Kaiso in breast tumorigenesis

Abstract Introduction: The canonical Wnt signalling pathway regulates various cellular processes (e.g. proliferation, differentiation, apoptosis) and leads to tumorigenesis when aberrantly activated. -catenin, the key downstream effector in Wnt signalling, translocates to the nucleus upon Wnt pathway activation, and activates target genes involved in tumorigenesis. The transcription factor Kaiso is a novel BTB/POZ (Broad complex, Tramtrak, Bric à brac/Pox virus and zinc finger) zinc finger (POZ-ZF) protein that specifically interacts with the cell adhesion cofactor p120ctn. Recent studies from our lab and others have revealed that Kaiso negatively regulates the canonical Wnt pathway by repressing activation of Wnt/ -catenin target genes. Some studies implicate Kaiso as a tumor suppressor (decreased expression in breast, ovarian and colon tumors), and others implicate Kaiso in tumor initiation (Kaiso-deficient mice are resistant to intestinal cancer). The goal of this study is to elucidate the biological role of Kaiso in breast tumorigenesis using various cellular assays. Methods: To assess Kaiso's role in breast cancer, Kaiso was misexpressed in MCF7 (human breast adenocarcinoma) cell lines. Stable Kaiso overexpressing and depleted cell lines were generated and assessed for cell proliferation and cell migration using focus formation, wound healing and matrigel invasion assays. Western blot analysis was performed to assess the effects of Kaiso misexpression on p120 and E-cadherin as well as epithelial-mesenchymal transition markers such as N-cadherin, Snail, Twist and Vimentin. The subcellular localization of Kaiso and its putative target genes in vivo was examined using immunohistochemistry (IHC) analysis of matched pairs of human normal and tumor breast tissue. Results: Focus formation assays revealed increased transformation by cells over-expressing Kaiso, suggesting an oncogenic role for Kaiso in cell culture. Preliminary wound healing and matrigel assays show increased cell motility and migration respectively in Kaiso over-expressing cells compared to parental control cells, thereby suggesting a role for Kaiso in the epithelial-mesenchymal transition. IHC analysis revealed predominant nuclear Kaiso expression in lobular and ductal breast carcinomas in breast tissues. These data support Kaiso's potential role as an oncogene in human breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-271.

RETRACTED: Kaiso is expressed in lung cancer: Its expression and localization is affected by p120ctn

Kaiso is a recently identified transcription factor that binds to p120-catenin (p120ctn), an Armadillo catenin and cell adhesion cofactor. However, clinical studies of human solid tumors have not been reported to investigate relationship between these proteins. Expression and localization of Kaiso and p120ctn were examined in 196 lung cancer specimens (including 55 cases of paired lymph node metastases and 80 cases with complete follow-up records) by immunohistochemistry. Three lung cancer cell lines, BE1, SPC, and A549 were used to establish p120ctn stably ablated or overexpressed cell lines. Co-immunoprecipitation was used to confirm p120ctn bind Kaiso in lung cancer tissue and cell lines. Localization and expression levels of Kaiso were detected via immunofluorescence, cytoplasmic vs. nuclear fractionation Western blot analysis and reverse transcription-polymerase chain reaction. Cytoplasmic Kaiso expression was evident in 115 (58.7%), and abnormal p120ctn expression was noted in 168 (85.7%). Cytoplasmic Kaiso and abnormal p120ctn expressions were associated with higher degree of malignancy (high-stage and lymph node metastases, all P<0.05). Abnormal p120ctn and cytoplasmic Kaiso expressions were higher in matched autologous nodal metastases than in primary growths. The lung cancer-related 5-year survival rate was significantly lower in patients who were cytoplasmic Kaiso-positive (22.9%; P=0.029) or abnormal p120ctn expression (20.6%; P=0.001). Multivariate analysis showed abnormal p120ctn expression was an independent factor defining the clinicopathological characters of patients. Cytoplasmic Kaiso expression was correlated with cytoplasmic p120ctn, they formed Kaiso-p120ctn complex in lung cancer tissues and cell lines. In addition, p120ctn ablation and overexpression altered Kaiso subcellular localization and protein level. Although both isoforms can regulate subcellular localization and protein levels of Kaiso, we found that only p120ctn isoform 3, but not isoform 1, directly interacts with Kaiso. p120ctn and Kaiso might co-participate in the progression and lymph node metastasis of lung cancer. p120ctn regulates expression and localization of Kaiso in lung cancer cells.