High E-cadherin Expression Research Articles

Establishment of human pluripotent stem cell-derived cortical neurosphere model to study pathomechanisms and chemical toxicity in Kleefstra syndrome

In the present study, we aimed to establish and characterize a mature cortical spheroid model system for Kleefstra syndrome (KS) using patient-derived iPSC. We identified key differences in the growth behavior of KS spheroids determined by reduced proliferation marked by low Ki67 and high E-cadherin expression. Conversely, in the spheroid-based neurite outgrowth assay KS outperformed the control neurite outgrowth due to higher BDNF expression. KS spheroids were highly enriched in VGLUT1/2-expressing glutamatergic and ChAT-expressing cholinergic neurons, while TH-positive catecholamine neurons were significantly underrepresented. Furthermore, high NMDAR1 expression was also detected in the KS spheroid, similarly to other patients-derived neuronal cultures, denoting high NMDAR1 expression as a general, KS-specific marker. Control and KS neuronal progenitors and neurospheres were exposed to different toxicants (paraquat, rotenone, bardoxolone, and doxorubicin), and dose-response curves were assessed after acute exposure. Differentiation stage and compound-specific differences were detected with KS neurospheres being the most sensitive to paraquat. Altogether this study describes a robust 3D model system expressing the disease-specific markers and recapitulating the characteristic pathophysiological traits. This platform is suitable for testing developing brain-adverse environmental effects interactions, drug development, and screening towards individual therapeutic strategies.

The Protective Potential of Butyrate against Colon Cancer Cell Migration and Invasion Is Critically Dependent on Cell Type.

Short-chain fatty acids such as butyrate are produced through the fermentation of dietary fiber by colonic bacteria. Preclinical studies indicate an anticancer potential of butyrate, but clinical evidence shows greater variability. The study hypothesizes the effectiveness of butyrate on reducing colon cancer cell migration and invasion may vary due to the cell-type. The study determines the efficacy of butyrate (0-4mM) to inhibit cancer cell migration, invasion, and related signaling proteins in three distinct human colorectal cancer (CRC) cell lines: HCT116, HT-29, and Caco-2.Butyrate exhibits a dose-dependent inhibitory effect on cancer cell migration and invasion. This inhibitory potential on oncogenic focal adhesion kinase (FAK) and sarcoma (Src) proteins is greater in HCT116 cells (1.1 and 0.8-fold) and HT-29 cells (0.9 and 0.4-fold) compared to Caco-2 cells, respectively. Conversely, E-cadherin protein, a classical epithelial cell marker and potential tumor suppressor, is 2.3-fold greater in HCT116 cells than in HT-29 cells and Caco-2 cells. Moreover, survival analysis from a public cancer database demonstrates that CRC patients with high E-cadherin expression have a 13% greater 5-year survival rate than those with low expression. Collectively, butyrate's anti-cancer efficacy on CRC cells varies depending on cell-type and is linked to the FAK/Src/E-cadherin pathway.

Expression of miR-34a, RASSF1A and E-cadherin in relation to PRB in endometrioid carcinoma and its precursor.

The current study aims to evaluate the levels of miR-34a, RASSF1A, and E-cadherin in relation to the levels of isoform B of progesterone receptor (PRB) in endometrioid carcinoma (EC) and atypical hyperplasia (AEH) and their association with clinicopathological parameters. 105 cases (35 EC, 35 AEH, and 35 control) were involved in this study. Cases of AEH received treatment, and other samples were obtained after 6 months to assess the response. E-cadherin and PRB were assessed by immunohistochemistry (IHC), RASSFA methylation by MSP-PCR, and its serum level by ELISA and miR-34a via quantitative PCR. The expressions of miR-34a, RASSF1A, E-cadherin, and PRB differ among the studied groups; all were higher in normal compared with AEH and EC, with a statistically significant difference. The higher PRB expression and decreased miR-34a and RASSF1A expression were associated with resistance to hormonal therapy in AEH. High PRB in EC is associated with lower RASSFA1, E-cadherin, and miR-34a. Decreased expressions of RASSF1A, miR-34a, and E-cadherin had a significant connection to advanced stages. Expression of PRB and miR-34a and serum levels of RASSF1A predict response to treatment in cases of AEH. High PRB and low E-cadherin expression are associated with progressive disease in EC.

Kinesin Facilitates Phenotypic Targeting of Therapeutic Resistance in Advanced Prostate Cancer.

Understanding the mechanisms underlying resistance is critical to improving therapeutic outcomes in patients with metastatic castration-resistant prostate cancer. Previous work showed that dynamic interconversions between epithelial-mesenchymal transition to mesenchymal-epithelial transition defines the phenotypic landscape of prostate tumors, as a potential driver of the emergence of therapeutic resistance. In this study, we use in vitro and in vivo preclinical MDA PCa patient-derived xenograft models of resistant human prostate cancer to determine molecular mechanisms of cross-resistance between antiandrogen therapy and taxane chemotherapy, underlying the therapeutically resistant phenotype. Transcriptomic profiling revealed that resistant and sensitive prostate cancer C4-2B cells have a unique differential gene signature response to cabazitaxel. Gene pathway analysis showed that sensitive cells exhibit an increase in DNA damage, while resistant cells express genes associated with protein regulation in response to cabazitaxel. The patient-derived xenograft model specimens are from patients who have metastatic lethal castration-resistant prostate cancer, treated with androgen deprivation therapy, antiandrogens, and chemotherapy including second-line taxane chemotherapy, cabazitaxel. Immunohistochemistry revealed high expression of E-cadherin and low expression of vimentin resulting in redifferentiation toward an epithelial phenotype. Furthermore, the mitotic kinesin-related protein involved in microtubule binding and the SLCO1B3 transporter (implicated in cabazitaxel intracellular transport) are associated with resistance in these prostate tumors. Combinational targeting of kinesins (ispinesib) with cabazitaxel was more effective than single monotherapies in inducing cell death in resistant prostate tumors. Implications: Our findings are of translational significance in identifying kinesin as a novel target of cross-resistance toward enhancing therapeutic vulnerability and improved clinical outcomes in patients with advanced prostate cancer.

Association of E-Cadherin Expression with Histological Type and Grade of Gastric Adenocarcinoma

Objectives: To determine the immune expression of E-Cadherin in gastric adenocarcinomas and its association with tumor histological type and grade to evaluate its prognostic significance. Methodology: This descriptive cross-sectional study was conducted at the Pakistan Institute of Medical Sciences Islamabad, from December 2017 to January 2021. Eighty-one patients from both genders, aged 21 to 83 years, undergoing gastric biopsies and Gastrectomy specimens to diagnose gastric adenocarcinoma were included in our study. Immunohistochemical processing was performed and the results were interpreted by the consultant pathologist for the expression of E-cadherin on gastric adenocarcinoma. The data were analyzed with SPSS version 17. Results: There was a significant statistical association between loss of E-Cadherin with histological type of adenocarcinoma (p = 0.000). In total 81 cases, there were 53 (65%) cases of intestinal type adenocarcinoma and 28 (35%) cases of diffuse type adenocarcinoma. Of the 28 cases with diffuse type gastric Adenocarcinoma, 10 cases showed complete loss of E-Cadherin. This complete loss of Cadherin was also seen in four out of 53 cases of intestinal adenocarcinoma, all of which were poorly differentiated intestinal type of adenocarcinomas. Total cases of well differentiated type of adenocarcinoma were nine; all of these showed high expression of E-Cadherin. Conclusion: There is a strong association of E-Cadherin expression with the histological type and grade of gastric adenocarcinoma according to Lauren’s classification. As the grade of the adenocarcinoma gets higher there is more loss of E-Cadherin expression which proves its prognostic significance. It was also established that gastric adenocarcinoma is a disease of elderly with male predominance.

The Immunohistochemical expressions and diagnostic values of E-cadherin and β-Catenin in suspected cervical lesion or cervical cancer

Background: Cervical lesions are the cellular changes of a pathologic process with a bacterial and viral etiology ranging from cervicitis, cervical intraepithelial neoplasm and invasive cancer. The integrity of the cervical epithelium is maintained by complex cell adhesion molecules. This work is aimed at determining the expression pattern diagnostic value of the adhesion molecules to enhance early detection of cancer. Methods: The study population was from 10 years retrospective archival tissue blocks of cervical cancer among women in Abuja metropolis from 2005 to 2015 Histopathological diagnosis made, from the 80 cases examined 20(25%) cases were cervicitis (CC), 15(18.8%) were cervical intraepithelial neoplasia (CINI), 15(18.8%) were cervical intraepithelial neoplasia (CIN2) while 30(37.3%) cases were squamous cell carcinoma SCC. They were examined immunohistochemically by Avidin Biotin Complex (ABC) method. E-Cadherin and Beta-Catenin, (cell adhesion molecules). Results: Statistically significant rate of positivity were found in the expression of β-catenin 100% among all the categories of the studied cases. β-catenin expression was significantly higher in CINI than in SCC, while in E-cadherin, therewere no statistically significant difference (p<0.05) between the distribution of high expression and low expression of E-cadherin in both CINI and SCC. In expression of E-cadherin among the cases studied, there was a significant difference (p<0.05) between rate of positivity and negativity for the expression of E-cadherin. A statistically significant rate of positivity was found in the expression of β-catenin among all the categories of the studied cases. In the expression E-cadherin, there was a gradual decrease in high expression from cervicitis to CINI, CIN2 and SCC. There was Significant (p<0.05) high degree of expression of E-cadherin in cervicitis and CINI against CIN2 and SCC, while a significant (p<0.05) low degree expression of E-cadherin in CIN2 and SCC against Cervicitis and CINI. In β-catenin expression, there as a gradual decrease in high expression from cervicitis to CIN2 and SCC expect in CINI were all had high expression. Conclusion: This study showed that most of the immunological markers examined can be used for identification of different stages involved in cancer progression, also for diagnosis of cancer, predictive prognosis and can be explore as therapy targets for cervicitis and Squamous cell carcinoma.

Predictors of long-term remission in acromegaly – an updated review

Abstract Acromegaly is a rare chronic disorder due to growth hormone (GH) - secreting pituitary tumor. The first-line treatment of acromegaly is a transsphenoidal adenomectomy. Surgical remission rates range from 30-50% for macroadenomas to 70-90% for microadenomas. Thus in patients with active acromegaly after surgery, medical treatment is recommended. Treatment outcome on medical therapy differs a lot depending on medications. The aim of this paper is to summerize the current knowledge of factors predicting acromegaly remission after surgery and biochemical control on medical treatment. Factors affecting treatment outcome, not related to the patient and the tumor, are neurosurgeon’s experience and visualisation tools used during surgery. Younger patients, as well as women tend to achieve acromegaly remission after surgery less often. Patients with lower GH and insulin-like growth factor-1 (IGF-1) concentrations at diagnosis, in oral glucose tolerance test (OGTT) and postoperatively have a greater chance of surgical remission. However, some publications do not confirm that. Patients presenting with larger and more invasive tumors achieve surgical remisson less often. Hypointensive tumors on T2-weighted magnetic resonance imaging (MRI) scans are associated with better response to first-generation somatostatin receptor ligands (SRLs) and to pasireotide. Pathologic factors increasing the chance of acromegaly remission are densely granulated tumors, lower Ki-67 index, high expression of somatostatin receptors (SSTRs) and high expression of E-cadherin. A thorough analysis of the abovementioned factors predicting the treatment outcome in acromegaly may be helpful for selecting the best treatment option for each patient.

E-cadherin expression in the tumor microenvironment of advanced epidermal growth factor receptor-mutant lung adenocarcinoma and the association with prognosis

BackgroundThe expression of programmed death-ligand 1 (PD-L1), tumor-infiltrating lymphocytes (TILs), E-cadherin, and vimentin in lung cancer tumor microenvironment is known to impact patient survival or response to therapy. The expression of these biomarkers may also differ between primary lung tumors and brain metastatic tumors. In this study, we investigated the interaction between these biomarkers in lung tumors with or without concomitant brain metastasis and the interaction with paired brain metastatic tumors.MethodsThe study included 48 patients with stage IV epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma. Sixteen of the forty-eight patients were diagnosed with brain metastasis, while the remaining thirty-two were not. All sixteen patients with brain metastasis had brain tumors. The expression of PD-L1, TILs (CD8+ T lymphocytes and FOXP3+ regulatory T lymphocytes), E-cadherin, and vimentin were evaluated using immunohistochemical (IHC) staining.ResultsPatients with brain metastasis exhibited a higher frequency of exon 19 deletion and uncommon EGFR mutations, a higher lung tumor vimentin score, worse progression-free survival (PFS), and overall survival (OS) than patients without brain metastasis. IHC staining showed no difference between paired lung and brain tumors. Patients with low PD-L1 expression had better PFS and OS. After multivariate analysis, higher body mass index, the presence of brain metastasis, bone metastasis, and uncommon EGFR mutations were correlated with worse PFS, while the presence of brain metastasis and high lung tumor E-cadherin score was associated with worse OS.ConclusionsIn patients with stage IV EGFR-mutant lung adenocarcinoma, high E-cadherin expression in the lung tumor might be associated with worse OS. Vimentin expression in the lung tumor was positively related to the risk of brain metastasis.

CANCER STEM CELLS IN ORAL CANCER: PHENOTYPIC COMPARISON BETWEEN SUBPOPULATIONS LABELED WITH CD44 AND CD326 ANTIBODIES.

Objective Cancer stem cells (CSC) comprise a subpopulation of tumor cells associated with initiation, progression, and chemoresistance. Subpopulations of CSC were characterized in a metastatic cell line LN1-A derived from the oral squamous cell carcinoma (OSCC), previously isolated in our laboratory. Study Design We selected three subpopulations of CSC (CD44Low/CD326+, CD44+/CD326-, and CD44High/CD326-) among the seven phenotypes isolated by labeling CD44 and CD326 antibodies by flow cytometry in metastatic LN-1A cells. Cell morphology, proliferation, migration, adhesion, clonogenic assay, cell cycle, cell death, and cisplatin chemoresistance were characterized. Western blotting for EFGR, FASN, p-AKT, CPT1-α, and mesenchymal epithelium transition markers were performed. Results CD44Low/CD326+ subpopulations showed an epithelial phenotype with polygonal morphology and high E-cadherin expression, while CD44+/CD326- and CD44High/CD326- showed fusiform morphology with high Vimentin and Slug expressions, suggesting a mesenchymal phenotype. Proliferation was higher in CD44+/CD326- and CD44High/CD326- cells, which was accompanied by increased levels of p-AKT. Cell adhesion and colony formation were increased in CD44+/CD326- and CD44High/CD326-, while cell death was higher in CD44Low/CD326+. No significant differences in cell cycle, cisplatin resistance, and the expression of EGFR, FASN, and, CPT1-α were observed between phenotypes. Conclusion Our results showed that CD44+/CD326- and CD44High/CD326- cells show a more aggressive phenotype. Cancer stem cells (CSC) comprise a subpopulation of tumor cells associated with initiation, progression, and chemoresistance. Subpopulations of CSC were characterized in a metastatic cell line LN1-A derived from the oral squamous cell carcinoma (OSCC), previously isolated in our laboratory. We selected three subpopulations of CSC (CD44Low/CD326+, CD44+/CD326-, and CD44High/CD326-) among the seven phenotypes isolated by labeling CD44 and CD326 antibodies by flow cytometry in metastatic LN-1A cells. Cell morphology, proliferation, migration, adhesion, clonogenic assay, cell cycle, cell death, and cisplatin chemoresistance were characterized. Western blotting for EFGR, FASN, p-AKT, CPT1-α, and mesenchymal epithelium transition markers were performed. CD44Low/CD326+ subpopulations showed an epithelial phenotype with polygonal morphology and high E-cadherin expression, while CD44+/CD326- and CD44High/CD326- showed fusiform morphology with high Vimentin and Slug expressions, suggesting a mesenchymal phenotype. Proliferation was higher in CD44+/CD326- and CD44High/CD326- cells, which was accompanied by increased levels of p-AKT. Cell adhesion and colony formation were increased in CD44+/CD326- and CD44High/CD326-, while cell death was higher in CD44Low/CD326+. No significant differences in cell cycle, cisplatin resistance, and the expression of EGFR, FASN, and, CPT1-α were observed between phenotypes. Our results showed that CD44+/CD326- and CD44High/CD326- cells show a more aggressive phenotype.

Association of wnt signalling with metachronous colonic polyp risk.

232 Background: Following colorectal polypectomy, 20-50% of patients develop metachronous polyps, and a proportion have increased colorectal cancer (CRC) risk. However, there are currently no molecular biomarkers for predicting metachronous polyp risk. Constitutive activation of the Wnt signalling cascade is a hallmark of polyp development and carcinogenesis, therefore biomarkers from this pathway were investigated for association with detection of metachronous polyps. Methods: A retrospective study was performed on a tissue microarray (TMA) of left sided colonic polyps from 279 patients undergoing screening polypectomy (May’09-Dec’16) followed by surveillance colonoscopy (up to 6 years). Mutational analysis was performed using the GPOL cancer plus panel, protein expression by Immunohistochemistry and RNA-sequencing by TempO-Seq (BioClavis). Log rank statistics determined associations with time to detection of metachronous polyp. Index polyp number, histology and size were compared to metachronous lesion outcome using χ2/ANOVA and multivariate polynomial regression identified independent predictors of advanced future lesions. Results: APC (91%), KRAS (30%) and SOX9 (23%) were the most frequently mutated genes and APC and SOX9 are both implicated in Wnt signalling. Mutations in APC and co-mutations in SOX9 with ARID1 were associated with a shorter time to detection of metachronous polyps. Neither variant classification nor location of mutations were associated with metachronous polyp detection. Luminal epithelial cells with high expression of E-Cadherin or SOX9 were significantly associated with earlier detection of metachronous polyps or CRC (HR 2.3, 95%CI 1.6-3.3: p<0.001 and: HR 2.0, 95%CI 1.3-3.0; p=0.001 respectively). By χ2/ANOVA analysis, E-Cadherin and SOX9 expression was associated with detection of metachronous polyp (p>0.001, p=0.037 respectively). On multivariate regression, number of polyps (HR 1.2, 95%CI 1.1-1.3; p<0.001), and E-Cadherin expression (HR 2.1, 95%CI 1.5-3; p<0.001), independently predicted metachronous lesions. Gene set enrichment analysis (GSEA) was used to explore differential biology between high and low E-Cadherin or SOX9 expression. For E-Cadherin, all seven immune-annotated MSigDB hallmarks had negative enrichment, however Wnt--catenin signaling was positively enriched (NES=1.50, p-adj<0.1). The top signature enriched for SOX9 was ‘NFB signaling via TNF (NES=1.70, p-adj<0.001). The LGR5 stemness signature was enriched in both high E-Cadherin (NES=1.97 p-adj<0.0001) and high SOX9 (NES=1.48, p-adj<0.0001) groups. Conclusions: Wnt signalling-associated protein expression within index polyp tissue is valuable for predicting metachronous polyp risk. In addition, expression of E-Cadherin and SOX9 in colonic polyps are linked to immune and stemness signatures, suggesting immune and stemness biology may be an underlying driver of metachronous polyp development.

ISOLATION AND CHARACTERIZATION OF CANCER STEM CELL SUBPOPULATIONS IN ORAL SQUAMOUS CELL CARCINOMA

<h3>Objectives</h3> Cancer stem cells (CSCs) comprise a subpopulation of tumor cells associated with initiation, progression, and chemoresistance. We identified, isolated, and characterized subpopulations of CSC in SCC-9 ZsGreen (SCC-9 ZsG) and LN1-A cell lines derived from oral squamous cell carcinoma (OSCC). <h3>Study Design</h3> SCC-9 ZsG and LN-1A subpopulations were identified and isolated with antibodies against CD44 and CD326 by flow cytometry. Cell morphology, proliferation, migration, clonogenic assay, and protein expression associated with epithelial-mesenchymal transition were carried out. <h3>Results</h3> Six and 7 phenotypes were isolated in SCC-9 ZsG and LN-1A cells in relation to its differential expression, respectively. CD44-low/CD326+ subpopulations showed epithelial phenotype with polygonal morphology and high expression of E-cadherin, whereas CD44+/CD326− showed a fusiform morphology with a high expression of vimentin, N-cadherin, and Slug, suggesting a mesenchymal phenotype. CD44-high/CD326− subpopulations were exclusively identified and isolated in LN-1A cells, also presenting a mesenchymal phenotype. Proliferation, migration, and colony formation were increased in CD44+/CD326− compared to CD44-low/CD326+ and parental cells in SCC-9 ZsG. <h3>Conclusions</h3> Our preliminary results showed that CD44-high/CD326− cells were correlated to a more malignant phenotype. Further experiments will be needed to better characterize the populations of CSC in OSCC.

Immunohistochemical expression of E-Cadherin in gastric carcinoma in Erbil city, Kurdistan Region

Background and objectives: Gastric carcinoma is at fourth position incidence-wise and holds a second place for deaths due to cancer. The Cadherins represent an important class of the adhesiveness molecules, which play an essential role in the homotypical cell-cell adhesion and in the complex process of invasion and metastasis. The point of this study is to identify the rate of the E-Cadherin immunoexpression in gastric carcinoma in Erbil locality and investigate its association with the clinico pathological parameters. Methods: A retrospective study was carried out in Rizgary teaching hospital and private laboratories in Erbil city for the period from Jan.2018 - Jan.2020. A total of 75 formalin fixed, paraffin embedded archival tissue blocks of total or partial gastrectomy samples for gastric carcinoma cases were collected, then E-cadherin immunohistochemistry was tested. Results: Twenty-one cases (28%) were categorized as high expression for E-Cadherin immunoexpression. At the same time 38 (50.7%) of the cases were classified as low expression and 16 (21.3%) of cases were labelled with no expression. E-Cadherin status was significantly associated with the tumor type and tumor grade, in which high E-Cadherin expression about (61.9%) was demonstrated in intestinal type tumors, (38.1%) in diffuse type tumors and about (57.1%) was performed in well-moderately differentiated tumors and (42.9%) in poorly differentiated tumors. While no significant association was found between E-Cadherin expression and other clinico. Conclusions: Our result support the hypothesis that the E-Cadherin immunostain is commonly expressed in Gastric cancer, and it was significantly related with type and grade of the tumor.

Особливості експресії молекул клітинної адгезії Е-кадгерину в різних молекулярних підтипах інвазивного протокового раку грудної залози

The purpose of the study was to determine the role of E-cadherin in the development and progression of molecular subtypes of invasive ductal breast cancer by assessing the expression of E-cadherin in various clinical and pathological prognostic parameters. Materials and methods. We demonstrated a comprehensive morphological, including immunohistochemical study of 193 cases of invasive ductal breast cancer with the molecular phenotype definition. General histological processing of samples was performed according to standard methods. Immunohistochemical studies for E-cadherin, ER, PR, c-erbB2, Ki-67 were performed according to standardized analytically validated protocols with the necessary controls. The grade of malignancy was determined according to the modified scheme of P. Scarff, H. Bloom and W. Richardson. E-cadherin expression level was quantified in 86 observations using the Qureshi counting system. Comparison of E-cadherin expression in different clinical and pathological parameters was evaluated using Pearson's test χ2. For all types of analysis, differences were considered significant at p &lt;0.05. Results and discussion. E-cadherin expression in ductal breast cancer tissue is significantly lower in cases with lymph node metastases than without metastatic lymph node involvement (χ2 = 4.55, p = 0.031). Low expression of E-cadherin or its absence was associated with pT3 tumors, clinical stage 3, with G2 and G3 malignancies. Loss of E-cadherin expression has an unfavorable prognostic value. E-cadherin expression is associated with the molecular type of invasive breast ductal carcinoma. High E-cadherin expression was common in ER-positive tumors of the luminal A phenotype and was determined in patients of both premenopausal and postmenopausal age, suggesting that ER-positive expression may be involved in the regulation of E-cadherin expression. Low tumor activity of cells of invasive ductal breast cancer of the luminal subtype is accompanied by an increase in the adhesive properties of these cells due to the high level of expression of E-cadherin. E-cadherin is considered an independent marker of triple-negative breast cancer and is characterized by an unfavorable prognosis and short life expectancy. Triple-negative cancer was associated with a significant predominance of patients with low and negative E-cadherin expression (p = 0.011). Conclusion. Thus, E-cadherin is a potent tumor suppressor of breast cancer. According to this role in the progression of breast cancer, it was found that partial or complete loss of E-cadherin expression correlates with an unfavorable prognosis in patients

Anti‐Tumor Effect of Embryonic Stem Cell Derived Exosomes in Triple Negative Breast Cancer: Potential Role of TCF7‐E‐Cadherin and VEGF

BackgroundTriple Negative Breast Cancer (TNBC) is an aggressive form of cancer and accounts for 10‐20 % of all breast cancer types. TNBC patients are widely treated with chemotherapeutic drugs such as doxorubicin (Dox), which is a primary standard of care. Dox treatment results in unwanted side effects including cardiac dysfunction and skeletal muscle weakness. We previously demonstrated the anti‐tumor effects and cardioprotective effects of embryonic stem cell derived exosomes (Es‐Exos) in the xenograft model of TNBC. The purpose of this study was to identify the biomolecules present in Es‐Exos, which may be linked to the anti‐tumor effects in the xenograft model.Methods and ResultsOrthotopic xenograft model of TNBC was generated by injecting human MDA‐MB‐231‐Tom‐Luc cell line in mammary intraductal T4 region of nude mice. Non‐tumor bearing female mice were used as control (NT‐Con;n=6) as applicable. The animals (n=10/group) were randomized to receive Saline (Con); Dox (5 mg/kg/week); Es‐Exos (50 ug/animal/twice a week; Dox+Es‐Exos; Es‐Exos alone and MEF‐Exos‐(mouse embryonic fibroblast cell derived exosomes 50 ug/animal/twice a week) for up to 4 weeks. Skeletal muscle grip strength declined both in Con and upon treatment with Dox in tumor bearing mice as compared to NT‐Con, which was restored upon treatment with Es‐Exos. Cardiac function measured by echocardiography showed decline in ejection fraction (EF) in saline treated tumor bearing Con and in Dox group, when compared to NT‐Con. Treatment with Dox+Es‐Exos and Es‐Exos improved ejection fraction compared to Con and Dox cohort. Treatment with Es‐Exos significantly reduced the tumor size compared to Dox and saline treated Con groups (Con Vs Dox and Dox+Es‐Exos; p‐value &lt;0.05). Picro Sirius red staining showed increased deposition of collagen in saline treated Con, which was reduced in Dox+Es‐Exos and Es‐Exos groups. RNA seq analysis of Es‐Exos identified several anti‐proliferative genes including a 6‐fold expression of Transcription Factor 7 (TCF7) and 6.5‐fold higher expression of E‐Cadherin (CDH1). Immunohistochemistry analysis of tumor tissue showed abundant expression of VEGF in untreated Con, which was reduced by combined treatment with Es‐Exos and Dox.ConclusionEmbryonic stem cells derived exosomes contain anti‐proliferative genes which may possibly lead to their anti‐tumor property in the TNBC tumor model.

E-cadherin expressions on bladder and its association with cancer progressivity: a retrospective cohort study

BackgroundBladder cancer is characterized by high recurrence and progressivity. E-cadherin serves as one of the most important molecules involved in the epithelial cells’ cell-to-cell adherence, suggested to inhibit tumor cell progression. This study aims to investigate the association between the E-cadherin expressions with bladder cancer progressiveness in 3 years.MethodsThis study was a retrospective cohort study involving bladder cancer patients in Cipto Mangunkusumo Hospital, Jakarta. Diagnosis of bladder cancers was confirmed by histopathological and immunohistochemistry examination between 2011 and 2018, with both grading and staging determined by uropathologists and uro-oncologists. E-cadherin was examined through immunohistochemistry examination at the time of diagnosis. Data on demography, muscle invasion, clinical staging, grade, metastasis, multifocality, and recurrence were obtained from medical records and pathology reports. The association of E-cadherin expression to muscle invasion and non-muscle invasion bladder cancer was evaluated and statistically analyzed. Patients’ survival data were followed up by phone.ResultsForty bladder cancer patients with a mean age of 60.05 ± 10.3 years were included. Most subjects had high E-cadherin expression (85%), muscle invasion (65%), high grade (65%), no metastasis (87.5%), multifocality (65%), and no recurrence (62.5%). Lower expression of E-cadherin was associated with the higher clinical stage (p < 0.02) and metastasis (p < 0.001). Patients with low E-cadherin expression showed worse cumulative survival than the high one (mean 32 months vs. 25 months, p = 0.13).ConclusionsLow level of E-cadherin was associated with the higher risk of muscle invasion, clinical staging, histological grade, and risk of metastasis. Meanwhile, patients with the high level of E-cadherin showed a better three-year survival rate.

SOX9 Triggers Different Epithelial to Mesenchymal Transition States to Promote Pancreatic Cancer Progression.

Simple SummaryPancreatic cancers are lethal types of cancer. A majority of patients progress to an advanced and metastatic disease, which remains a major clinical problem. Therefore, it is crucial to identify critical regulators to help predict the disease progression and to develop more efficacious therapeutic approaches. In this work we found that an increased expression of the developmental factor SOX9 is associated with metastasis, a poor prognosis and resistance to therapy in pancreatic ductal adenocarcinoma patients and in cell cultures. We also found that this effect is at least in part due to the ability of SOX9 to regulate the activity of stem cell factors, such as BMI1, in addition to those involved in EMT and metastasis. Background: Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers mainly due to spatial obstacles to complete resection, early metastasis and therapy resistance. The molecular events accompanying PDAC progression remain poorly understood. SOX9 is required for maintaining the pancreatic ductal identity and it is involved in the initiation of pancreatic cancer. In addition, SOX9 is a transcription factor linked to stem cell activity and is commonly overexpressed in solid cancers. It cooperates with Snail/Slug to induce epithelial-mesenchymal transition (EMT) during neural development and in diseases such as organ fibrosis or different types of cancer. Methods: We investigated the roles of SOX9 in pancreatic tumor cell plasticity, metastatic dissemination and chemoresistance using pancreatic cancer cell lines as well as mouse embryo fibroblasts. In addition, we characterized the clinical relevance of SOX9 in pancreatic cancer using human biopsies. Results: Gain- and loss-of-function of SOX9 in PDAC cells revealed that high levels of SOX9 increased migration and invasion, and promoted EMT and metastatic dissemination, whilst SOX9 silencing resulted in metastasis inhibition, along with a phenotypic reversion to epithelial features and loss of stemness potential. In both contexts, EMT factors were not altered. Moreover, high levels of SOX9 promoted resistance to gemcitabine. In contrast, overexpression of SOX9 was sufficient to promote metastatic potential in K-Ras transformed MEFs, triggering EMT associated with Snail/Slug activity. In clinical samples, SOX9 expression was analyzed in 198 PDAC cases by immunohistochemistry and in 53 patient derived xenografts (PDXs). SOX9 was overexpressed in primary adenocarcinomas and particularly in metastases. Notably, SOX9 expression correlated with high vimentin and low E-cadherin expression. Conclusions: Our results indicate that SOX9 facilitates PDAC progression and metastasis by triggering stemness and EMT.

Local drug delivery system for the treatment of tongue squamous cell carcinoma in rats.

The present study describes a local drug delivery system with two functions, which can suppress tumor growth and accelerate wound healing. Thе system consists of a two-layer multicomponent fibrin-based gel (MCPFTG). The internal layer of MCPFTG, which is in direct contact with the wound surface, contains cisplatin placed on a CultiSpher-S collagen microcarrier. The external layer of MCPFTG consists of a CultiSpher-S microcarrier with lyophilized bone marrow stem cells (BMSCs). The efficacy of MCPFTG was evaluated in a rat model of squamous cell carcinoma of the tongue created with 4-nitroquinoline 1-oxide. The results of the study showed that, within 20–25 days, a non-healing wound of the tongue was formed in animals that underwent only 85% resection of squamous cell carcinoma, while rapid progression of the residual tumor was concomitantly observed. Immunohistochemical methods revealed high expression of cyclin D1 and low expression of E-cadherin in these animals. Additionally, high expression of p63 and Ki-67 was noted. In 80% of animals with squamous cell carcinoma of the tongue that were treated with MCPFTG after 85% tumor resection, a noticeable suppression of tumor growth was evident throughout 150 days, and tumor recurrence was not detected. Immunohistochemistry revealed low or moderate expression of cyclin D1, and high expression of E-cadherin throughout the whole observation period. The MCPFTG-based local drug delivery system was shown to be effective in suppressing tumor growth and preventing recurrence. MCPFTG decreased the toxicity of cisplatin and enhanced its antitumor activity. In addition, lyophilized paracrine BMSC factors present in MCPFTG accelerated wound healing after tumor removal. Thus, the present study suggests novel opportunities for the development of a multifunctional drug delivery system for the treatment of squamous cell carcinoma.

Efficacy, Safety, and Tumor Marker Inhibition of Apatinib Combined with Conventional Chemotherapy Regimens for Patients with Advanced Triple-Negative Breast Cancer.

Objective Triple-negative breast cancer (TNBC) is an aggressive disease with highly invasive nature and poor outcomes. Due to the absence of specific treatment strategies for this tumor subgroup, patients with TNBC are treated with conventional therapeutics, frequently leading to systemic relapse. In this study, we sought to investigate apatinib combined with conventional chemotherapy regimens in treating patients with advanced TNBC concerning the efficacy, safety, expressions of tumor markers, and patient survival. Methods This is a prospective study including 150 cases of advanced TNBC who were randomly arranged into a conventional group and combined group, with 75 cases per group. The patients in the conventional group were treated with conventional chemotherapy, and those in the combined group were treated with apatinib combined with conventional chemotherapy. The peripheral blood was collected from each patient, and carcinoembryonic antigen (CEA), carbohydrate antigen 153 (CA153), and carbohydrate antigen 125 (CA125) were determined. The expressions of nuclear proliferation antigen marker (Ki67), β-catenin, and E-cadherin were determined in the biopsy collected from each patient. Results The objective remission rate (ORR) and disease control rate (DCR) (41.33% and 81.33%) in the combined group were notably higher than those in the conventional group (29.33% and 68.00%) (P < 0.05). After treatment, the serum levels of CEA, CA153, and CA125 and the expressions of Ki67 and β-catenin were declined, but the expression of E-cadherin was increased in both groups; the combined group exhibited lower serum levels of CEA, CA153, and CA125, and the expressions of Ki67 and β-catenin were concurrent with a higher expression of E-cadherin than the conventional group (P < 0.05). No significant difference was noted between the two groups regarding the occurrence of adverse reactions (P > 0.05). Improved progression-free survival (PFS) was observed in the combined group compared to the conventional group (P < 0.05. Conclusion These findings suggest that apatinib combined with conventional chemotherapy regimens confers a prolonged PFS for treating patients with advanced TNBC.

E-Cadherin Regulates Mitochondrial Membrane Potential in Cancer Cells.

Simple SummaryCancer cells have unusually high mitochondrial membrane potential (ΔΨm). However, the microenvironmental mechanisms that regulate cancer cell ΔΨm remain unclear. In this study, we use in vitro micropatterned tumor models to mimic the confinement cues in tumor microenvironments and show that the E-cadherin mediated intercellular adhesion negatively regulates cancer cell ΔΨm.Epithelial cancer cells often have unusually higher mitochondrial membrane potential (ΔΨm) than their normal counterparts, which has been associated with increased invasiveness in vitro and higher metastatic potential in vivo. However, the mechanisms by which ΔΨm in cancer cells is regulated in tumor microenvironment (TME) remain unclear. In this study, we used an in vitro micropatterning platform to recapitulate biophysical confinement cues in the TME and investigated the mechanisms by which these regulate cancer cell ΔΨm. We found that micropatterning resulted in a spatial distribution of ΔΨm, which correlated with the level of E-cadherin mediated intercellular adhesion. There was a stark contrast in the spatial distribution of ΔΨm in the micropattern of E-cadherin-negative breast cancer cells (MDA-MB-231) compared to that of the high E-cadherin expressing (MCF-7) cancer cells. Disruption and knockout of E-cadherin adhesions rescued the low ΔΨm found at the center of MCF-7 micropatterns with high E-cadherin expression, while E-cadherin overexpression in MDA-MB-231 and MCF-7 cells lowered their ΔΨm at the micropattern center. These results show that E-cadherin plays an important role in regulating the ΔΨm of cancer cells in the context of biophysical cues in TME.

AdipoSIGHT in Therapeutic Response: Consequences in Osteosarcoma Treatment.

Chemotherapeutic resistance is a major problem in effective cancer treatment. Cancer cells engage various cells or mechanisms to resist anti-cancer therapeutics, which results in metastasis and the recurrence of disease. Considering the cellular heterogeneity of cancer stroma, the involvement of stem cells is reported to affect the proliferation and metastasis of osteosarcoma. Hence, the duo (osteosarcoma: Saos 2 and human adipose-derived stem cells: ASCs) is co-cultured in present study to investigate the therapeutic response using a nonadherent, concave surface. Staining with a cell tracker allows real-time microscopic monitoring of the cell arrangement within the sphere. Cell–cell interaction is investigated by means of E-cadherin expression. Comparatively high expression of E-cadherin and compact organization is observed in heterotypic tumorspheres (Saos 2–ASCs) compared to homotypic ones (ASCs), limiting the infiltration of chemotherapeutic compound doxorubicin into the heterotypic tumorsphere, which in turn protects cells from the toxic effect of the chemotherapeutic. In addition, genes known to be associated with drug resistance, such as SOX2, OCT4, and CD44 are overexpressed in heterotypic tumorspheres post-chemotherapy, indicating that the duo collectively repels the effect of doxorubicin. The interaction between ASCs and Saos 2 in the present study points toward the growing oncological risk of using ASC-based regenerative therapy in cancer patients and warrants further investigation.