P63 Expression Research Articles

Podoplanin Expression Independently and Jointly with Oral Epithelial Dysplasia Grade Acts as a Potential Biomarker of Malignant Transformation in Oral Leukoplakia.

Our aim was to evaluate the expression of biomarkers, CD44v6, CD147, EGFR, p53, p63, p73, p16, and podoplanin in oral leukoplakias (OL) and to assess their potential for prediction of malignant transformation (MT). We analyzed the expression of CD44v6, CD147, EGFR, p53, p63, p73, p16, and podoplanin by immunohistochemistry in 52 OL, comprised of 41 low-grade (LG) dysplasia and 11 high-grade (HG) cases. Twelve healthy normal tissues (NT) were also included. Univariate and multivariate analysis were performed to evaluate any association with MT. Variable expression among the studied markers was observed, with a significant increase of high expression from NT to LG and HG cases in CD44v6 (p = 0.002), P53 (p = 0.002), P73 (p = 0.043), and podoplanin (p < 0.001). In multivariate analysis, cases with high podoplanin score showed a significant increased risk of MT (HR of 10.148 (95% CI of 1.503–68.532; p = 0.017). Furthermore, podoplanin combined with binary dysplasia grade obtained a HR of 10.238 (95% CI of 2.06–50.889; p = 0.004). To conclude, CD44v6, p53, p73, and podoplanin showed an increasing expression along the natural history of oral carcinogenesis. Podoplanin expression independently or combined with dysplasia grade could be useful predictive markers of MT in OL.

Clinical outcomes associated with expression of aurora kinase and p53 family members in muscle-invasive bladder cancer.

Biomarkers are needed in muscle-invasive bladder cancer (MIBC). We previously reported that high tumor aurora kinase (AURK) A expression identifies patients with MIBC with poor prognosis. Aberrant p53 expression has also been associated with poor outcomes in MIBC, though to the best of our knowledge, co-expression rates of p53 and aurora kinases have not been previously described in MIBC. As aurora kinase and p53 family members may co-regulate each other, the present study investigated whether tumor p53 or p63 protein expression influenced the prognostic value of AURKA in a pilot study of 50 patients with MIBC treated with curative intent. Immunohistochemistry for AURKA, AURKB, p53 and p63 were performed on archival pre-treatment tumor specimens and correlated with clinical outcomes in patients with MIBC who received neoadjuvant chemotherapy (NAC) prior to cystectomy. Baseline p53 [hazard ratio (HR) 1.46; 95% confidence interval (CI)=0.55-3.9; P=0.448) and p63 (HR 2.02; 95% CI=0.51-8.1; P=0.313) protein expression did not predict for overall survival (OS). Low p53 protein expression did not correlate with high AURKA (φ=0.190) or AURKB (φ=0.075) expression. However, in tumors with low p53 expression (n=17), the presence of either high AURKA or AURKB expression levels predicted an increased risk for relapse (HR 27.1; 95% CI=2.7-270.1; P=0.005) and mortality (HR 14.9; 95% CI=2.3-95.6; P=0.004) compared to tumors with both low AURKA and AURKB levels. The relationship between p63 and AURKA/B expression levels was not tested due to the prevalence (80%) of high p63 expression in the present cohort. In tumors with low AURKA expression, p53 status did not predict for OS (HR 0.62; 95% CI 0.2-3.2; P=0.572). In multivariable analysis, only high baseline AURKA expression predicted for inferior OS (HR 4.9; 95% CI 1.7-14.1; P=0.003). To the best of our knowledge, the present study was the first to report co-expression of p53 and aurora kinase family members in MIBC, and although wild-type p53 may regulate the aurora kinases in preclinical models, the adverse prognostic value of tumor AURKA overexpression was independent from baseline tumor p53 protein expression in the present cohort. AURKA remains an important prognostic biomarker in patients with MIBC and warrants further evaluation in prospective studies to validate whether baseline AURKA can identify patients that are unlikely to benefit from standard of care with NAC.

Expression of p63 in Cervical Intraepithelial Neoplasia (CIN) and Cervical Cancer

Background: Cervical cancer is a major public health problem worldwide. Persistent infection with High-Risk Human Papilloma Virus (HPV) has been the main cause of squamous intraepithelial neoplasia which in turn leads to invasive squamous cell carcinoma. p63 is necessary for the activation of HPV, epithelial proliferation and differentiation. It also regulates the expression of certain cell cycle regulators. It has been reported that, from CIN I to CIN III, p63 expression increases progressively from basal layer to surface. In squamous cell carcinoma, it is expressed throughout the entire thickness of the tumor. Thereby it plays a significant role in diagnosing cervical premalignant and malignant lesions.&#x0D; Objective: To evaluate the relationship of p63 expression with different grades of CIN &amp; invasive SCC.&#x0D; Method: Total 86 paraffin embedded tissue blocks of histopathologically diagnosed cases of CIN and cervical cancer were evaluated by immunohistochemical staining for p63 expression. The study was performed in Sir Salimullah Medical College, Dhaka (from March, 2018 to February, 2020). Statistical analyses were carried out by using SPSS version 22 for Windows. A descriptive analysis was performed for all data. Observations were indicated by frequencies and percentages. Statistical significance was set at “p” value &lt;0.05.&#x0D; Results: Present study showed progressive increase in p63 expression from CIN I to CIN III from basal layer to surface. In invasive squamous cell carcinoma, higher expression of p63 was noted throughout the entire thickness of the tumor. No expression was seen in cervical adenocarcinoma and small cell carcinoma. In adenosquamous carcinoma only the area showing squamous differentiation revealed positive p63 expression. Statistically significant association of p63 expression was found with parity of patients and among grades of CIN .&#x0D; Conclusion: The results of this current study revealed that, p63 has significant association among different grades of CIN. It is also a useful marker in confirming a poorly differentiated squamous cell carcinoma &amp; predicting the progression of a squamous neoplastic lesion from cervical intraepithelial neoplasia to invasive squamous cell carcinoma. Moreover, it is useful to differentiate invasive squamous cell carcinoma from cervical adenocarcinoma.&#x0D; Sir Salimullah Med Coll J 2021; 29(2): 141-146

AKT2 identified as a potential target of mir-29a-3p via microRNA profiling of patients with high proliferation lacrimal gland adenoid cystic carcinoma

The lacrimal gland adenoid cystic carcinoma (LACC) is a major orbital malignancy. The recurrence rate and mortality rate are higher in high proliferation LACC(HP-LACC) compared with low proliferation LACC(LP-LACC). In this study, miRNA microarray was used to explore the differentially expressed miRNAs profiling between HP-LACC and LP-LACC and its potential signaling pathway. Tissues from 17 patients with LACC were collected and made into tissue microarrays. Patients were divided into a high proliferation group and a low proliferation group based on Ki-67 value. HE, immunofluorescence (IF), and Immunohistochemistry (IHC) were performed on the tissue microarrays. Eight LACC tissues(4 HP-LACC and 4 LP-LACC) were made into miRNA microarrays and analyzed for miRNA profiles. Differentially expressed miRNAs were analyzed by volcano plot and heat map. Target gene were predicted using the miRWalk and miRDB for these differentially expressed miRNAs, the intersection of the results are used as targets for further gene ontology and KEGG pathway analysis.The four differentially expressed miRNAs were validated by qRT-PCR, the miRNAs with statistically significant differences validated by dual luciferase reporter and qRT-PCR. Finally, IHC was used for their downstream signaling pathway proteins.HE staining showed the presence of tubular, cribriform, and basaloid structures in LACC. IF showed the presence of CK7,P63 fluorescence expression in all three structures.Patients were divided into HP-LACC and LP-LACC based on Ki-67 median value of 11%. IHC and survival analysis showed with the increase of KI-67 ratio, the proportion of P63 decreased, and the expression of P53 increased. The disease-free survival and overall survival of the patients decreased. IHC and survival analysis showed as Ki-67 expression increased, P63 expression decreased, P53 expression elevated, with prognosis worse. Heat map and volcano plot yielded 15 differentially expressed miRNAs between HP-LACC and LP-LACC.The 15 differential miRNAs were used to predict target genes in miRWalk and miRDB databases respectively, and there were 559 target genes after intersection.559 predicted target genes obtained. Go and KEGG analysis showed that these target genes exerted important biological functions through multiple signaling pathways. Among the 15 differentially expressed miRNAs, miR-29a-3p was verified to be significant by qRT-PCR. Dual luciferase reporter and tissue microarray immunohistochemical assays validated that AKT2 was a direct target gene of miR-29a-3p. Current studies have identified differentially expressed miRNAs associated with LACCs of variable proliferation ability, and found that AKT2 is a direct target gene of miR-29a-3p, which will contribute to target gene therapy in patients with high proliferation LACC in the future.

Lacrimal Gland Microenvironment Changes After Obstruction of Lacrimal Gland Ducts.

PurposeTo investigate microenvironment changes of the lacrimal gland after obstruction of lacrimal gland ducts.MethodsThe ducts of rat exorbital lacrimal gland were ligated by sutures for different durations. After that, the sutures in some animals were released, and they were observed for 21 days to evaluate the recovery of the lacrimal gland. Slit lamp and tear secretion test was performed to evaluate ocular surface and lacrimal gland function. The lacrimal gland and cornea were harvested and processed for hematoxylin and eosin staining, oil red O staining, LipidTOX staining, Masson staining, quantitative real time polymerase chain reaction, and immunofluorescence staining.ResultsAfter the lacrimal gland ducts were blocked, tear secretion and the weight of the lacrimal gland were reduced. Incidence of corneal neovascularization increased after seven days. Intraglandular ducts dilated and acini destroyed. Long-term ligation induced fibrosis and lipid accumulation of the lacrimal glands. Inflammatory cell infiltrated and inflammatory factors upregulated. Proliferative and apoptotic cells increased. Structure of myoepithelial cells and basement membrane was destroyed. The p63 expression increased whereas Pax6 expression decreased. After suture release, tear secretion and structure of acini could recover in less than seven days after ligation, with a decrease in inflammatory cell infiltration and fibrosis relief. Apoptotic cells and proliferative cells increased at five days thereafter. The structure of the myoepithelial cells and basement membrane could not recover three days after ligation, and the number of mesenchymal cells increased in ligation after five to 14 days.ConclusionsBlockage of the lacrimal gland ducts results in dystrophy of lacrimal gland acini cells, inflammation, and lipid accumulation of the lacrimal gland microenvironment. Long-term duct blockage will cause irreversible lacrimal gland failure.

Primary Mucoepidermoid Carcinoma of the Lacrimal Apparatus

In this study, we evaluated the clinicopathologic and molecular characteristics of lacrimal apparatus mucoepidermoid carcinoma (MEC) to define its typical diagnostic features. Retrospective observational case series. Institutional pathology records between 2011 and 2021 were searched for all cases of lacrimal apparatus MEC. A total of 2 male and 6 female patients ranging in age from 18 to 83 years (median 56, mean 54) were included. Six lacrimal apparatus MECs were found in the lacrimal gland, and 2 cases occurred in the lacrimal sac and nasolacrimal duct. Histologically, there were 6 cases of conventional MEC, 1 clear-cell variant of MEC, and 1 oncocytic variant of MEC for a total of 8 cases. There were 3 low-grade cases and 5 high-grade cases. All 8 cases were evaluated via immunohistochemistry, and the results were positive (scores 1-4) for pankeratin, 34betaE12, p63, p40, CK7, CK8, and CK19, with a relatively higher expression of p63 observed in high-grade MEC. The presence of human papillomavirus (HPV) type 6 DNA was found in 4 patients. MAML2 fluorescence in situ hybridization was positive for MAML2 rearrangement in 3 lacrimal gland tumors (2 low-grade and 1 high-grade). Six tumors were managed with radical resection, and 2 patients underwent orbital exenteration. Postoperative radiation therapy was delivered to 6 patients, and chemotherapy was administered to 1 patient. MECs of the lacrimal apparatus are rare tumors, and the rate of MAML2 translocations is lower than that in salivary MECs. Lacrimal gland and lacrimal sac MECs may not be of the same subtypes intrinsically because of the difference in MAML2 translocation, anatomy, and clinical course. The etiologic function of HPV type 6 infection should be explored in lacrimal apparatus MECs. Radical surgery is the treatment of choice. The description of these unique findings may assist in the definitive diagnosis of and improve our understanding of lacrimal apparatus MEC.

Targeted deletion of TGFβ1 in basal keratinocytes causes profound defects in stratified squamous epithelia and aberrant melanocyte migration

Transforming Growth Factor Beta 1 (TGFβ1) is a multifunctional cytokine that regulates proliferation, apoptosis, and epithelial-mesenchymal transition of epithelial cells. While its role in cancer is well studied, less is known about TGFβ1 and regulation of epithelial development. To address this, we deleted TGFβ1 in basal keratinocytes of stratified squamous epithelia. Newborn mice with a homozygous TGFβ1 deletion had significant defects in proliferation and differentiation of the epidermis and oral mucosa, and died shortly after birth. Hair follicles were sparse in TGFβ1 depleted skin and had delayed development. Additionally, the Wnt pathway transcription factor LEF1 was reduced in hair follicle bulbs and nearly absent from the basal epithelial layer. Hemizygous knockout mice survived to adulthood but were runted and had sparse coats. The skin of these mice had irregular hair follicle morphology and aberrant hair cycle progression, as well as abnormally high melanin expression and delayed melanocyte migration. In contrast to newborn TGFβ1 null mice, the epidermis was hyperproliferative, acanthotic and inflamed. Expression of p63, a master regulator of stratified epithelial identity, proliferation and differentiation, was reduced in TGFβ1 null newborn epidermis but expanded in the postnatal acanthotic epidermis of TGFβ1 hemizygous mice. Thus, TGFβ1 is both essential and haploinsufficient with context dependent roles in stratified squamous epithelial development and homeostasis.

Immunomolecular Investigation of Human Papillomavirus Genotypes (16, 18) and P63 Expression in Patients with Malignant and Non-malignant Colorectal Tumors.

Cancer of the colon (colorectal cancer, or CRC) is the third most frequent malignancy in the world and the fourth leading cause of cancer-related death. Recent research has focused on the link between high-risk human papillomavirus (HPV) infections and the onset/development of several different types of cancer in humans. As a result, scientists are now paying more attention to HPV and CRC. In a variety of malignant tumors, P63 is overexpressed. This includes non-Hodgkin lymphoma and breast carcinoma, as well as lung, bladder, and prostate cancers. However, in accordance with the existence of many P63 isoforms in malignant tumors, the actions of P63 in these malignancies remain a source of debate. P63 immunohistochemistry expression in CRC tissues is being investigated as a possible etiological link between high-risk HPV types and CRC. This retrospective study intended to investigate if there was an etiological link between high-risk HPV types and CRC. It has utilized 92 chosen formalin-fixed and paraffin-embedded tissue block samples. The collected samples were divided into 62 blocks of colorectal adenocarcinoma mass tissues and 30 non-malignant colorectal tissues used as a control group. Chromogenic in situ hybridization (CISH) was employed to discover HPV DNA16/18 in colorectal tissues. The overall proportion of positive HPV16/18 DNA- CISH detection in the mass CRC group was 44.4%, whereas HPV16/18 DNA was obtained at 80.0% in the non-malignant control group. The overall proportion of positive P63-ISH detection in the CRC group was also 70.4%, whereas P63 was 73.3% in the non-malignant control group. The link between HPV infection and P63 expression in CRC might point to the importance of these molecules in the progression of CRC.

The c-Myc Oncogene Maintains Corneal Epithelial Architecture at Homeostasis, Modulates p63 Expression, and Enhances Proliferation During Tissue Repair.

PurposeThe transcription factor c-Myc (Myc) plays central regulatory roles in both self-renewal and differentiation of progenitors of multiple cell lineages. Here, we address its function in corneal epithelium (CE) maintenance and repair.MethodsMyc ablation in the limbal–corneal epithelium was achieved by crossing a floxed Myc mouse allele (Mycfl/fl) with a mouse line expressing the Cre recombinase gene under the keratin (Krt) 14 promoter. CE stratification and protein localization were assessed by histology of paraffin and plastic sections and by immunohistochemistry of frozen sections, respectively. Protein levels and gene expression were determined by western blot and real-time quantitative PCR, respectively. CE wound closure was tracked by fluorescein staining.ResultsAt birth, mutant mice appeared indistinguishable from control littermates; however, their rates of postnatal weight gain were 67% lower than those of controls. After weaning, mutants also exhibited spontaneous skin ulcerations, predominantly in the tail and lower lip, and died 45 to 60 days after birth. The mutant CE displayed an increase in stratal thickness, increased levels of Krt12 in superficial cells, and decreased exfoliation rates. Accordingly, the absence of Myc perturbed protein and mRNA levels of genes modulating differentiation and proliferation processes, including ΔNp63β, Ets1, and two Notch target genes, Hey1 and Maml1. Furthermore, Myc promoted CE wound closure and wound-induced hyperproliferation.ConclusionsMyc regulates the balance among CE stratification, differentiation, and surface exfoliation and promotes the transition to the hyperproliferative state during wound healing. Its effect on this balance may be exerted through the control of multiple regulators of cell fate, including isoforms of tumor protein p63.

P63 targeted deletion under the FOXN1 promoter disrupts pre-and post-natal thymus development, function and maintenance as well as induces severe hair loss.

Progressive immune deficiency of aging is characterized by severe thymic atrophy, contracted T cell repertoire, and poor immune function. p63 is critical for the proliferative potential of embryonic and adult stem cells, as well as thymic epithelial cells (TECs). Because p63 null mice experience rapid post-natal lethality due to epidermal and limb morphogenesis defects, studies to define a role for p63 expression in TEC biology focused on embryonic thymus development and in vitro experiments. Since post-natal thymic stromal development and function differs from that of the embryo, we assessed the impact of lineage-restricted p63 loss on pre- and post-natal murine TEC function by generating mice with a loss of p63 function targeted to TEC, termed p63TECko mice. In adult p63TECko mice, severe thymic hypoplasia was observed with a lack in a discernable segregation into medullary and cortical compartments and peripheral T cell lymphopenia. This profound thymic defect was seen in both neonatal as well as embryonic p63TECko mice. In addition to TECs, p63 also plays in important role in the development of stratified epithelium of the skin; lack of p63 results in defects in skin epidermal stratification and differentiation. Interestingly, all adult p63TECko mice lacked hair follicles despite having normal p63 expression in the skin. Together our results show a critical role of TEC p63 in thymic development and maintenance and show that p63 expression is critical for hair follicle formation.

Profile of Histopathological Type and Molecular Subtypes of Mammary Cancer of DMBA-induced Rat and its Relevancy to Human Breast Cancer

BACKGROUND: Animal models with mammary cancer that closely mimic human breast cancer for treatment development purposes are still required. Induction of 7,12-dimethylbenzanthracene (DMBA) to rats shows the histopathological features and mammary cancer characterization similar to humans. Examinations of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki67 expressions are crucial in deciding the treatment and prognosis of breast cancer. AIM: This research aimed to view histopathology images of mammary glands and expressions of ER, PR, Ki67, and HER2 of DMBA-induced rats. METHODS: After 1-week adaptation, 11 5-weeks-old female rats were induced with 20 mg/kg body weight (BW) of DMBA 2 times a week for 5 weeks. On week 29, nodules taken from the mammary gland were examined for hematoxylin-eosin staining and immunohistochemistry with p63, ER, PR, HER2, and Ki67 antibodies. The grading score used the Nottingham Grading System and molecular classifications based on St. Gallen 2013. RESULTS: Six rats had nodules, but the histopathologic features of one nodule showed normal mammary gland without cancer. The histopathological type of mammary cancer was cribriform carcinoma, comedo carcinoma, lipid-rich carcinoma, adenocarcinoma squamous, and adenomyepithelioma. Histopathological grading showed 60% of grade 3 and 40% of grade 2. P63 expression showed 60% positive and 40% negative. The frequency of ER, PR, HER2, and Ki67 of five nodules showed positivity: 40%, 60%, 60%, and 60%, respectively. Molecular subtypes of Luminal A, B, HER2, and triple-negative were 0%, 60%, 20%, and 20%, respectively. CONCLUSION: Histopathological features and molecular subtype of mammary cancer on rats induced with 20 mg/kg BW of DMBA showed similarity to human breast cancer.

Expression of Ki-67, P63, P40, and alpha-smooth muscle actin in salivary gland carcinomas with or without myoepithelial differentiation

Background: Myoepithelial cells are involved in the development of salivary glands. Many studies propose that these cells can prevent cell proliferation. Aim: This study aimed to investigate the expression of Ki-67, P63, P40, and alpha-smooth muscle actin (α-SMA) in salivary gland carcinomas with or without myoepithelial differentiation. Methods: A panel of myoepithelial markers including P63, P40, α-SMA, and Ki-67 were used for immunohistochemical study in 67 salivary gland carcinomas (33 with and 34 without myoepithelial differentiation). The percentage of positive cells was calculated (in high-power field) from a minimum of 1000 neoplastic cells. SPSS software (version 21) was used. Results: There was no statistically significant difference between Ki-67 expression and the presence or absence of myoepithelial cells (P = 0.6), but Ki-67 expression was related to the age (P = 0.032) and location of carcinomas (P = 0.001). All carcinomas with myoepithelial differentiation exhibited consistent P63+/P40+ staining, whereas polymorphous adenocarcinomas showed P63+/P40− immunophenotype. The expression of Ki-67 in adenoid cystic carcinomas was higher than mucoepidermoid carcinomas (P = 0.020) and polymorphous adenocarcinomas (P = 0.002). Conclusion: In the present study, although the decrease in the number of myoepithelial cells was associated with increased proliferation in adenoid cystic carcinomas, no such relationship was found in the overall assessment between the two groups. This can be justified by the fact that the clinical behavior of salivary carcinomas and their cell proliferation may be affected by factors other than the presence of myoepithelial cells or lack thereof. Ki-67 and P63/P40 expressions may be useful to differentiate adenoid cystic carcinomas from polymorphous adenocarcinomas in small biopsies.

Association of human papillomavirus and Epstein-Barr virus with squamous cell carcinoma of upper aerodigestive tract.

Cancers of the upper aero-digestive tract (UADT) are fifth most common cancer in the world with around 10,55,000 new cases and 7,25,000 deaths worldwide. Tobacco and alcohol act synergistically and are the two most important etiological factors responsible for about 75% of SCC. Studies have reported changing trends in the incidence of SCC showing an increasing shift in epidemiology attributed to the infection by viruses. The most commonly implicated viruses are the Human Papillomavirus (HPV) and Epstein-Barr virus (EBV). To study association of Human Papilloma virus (HPV) and Epstein Barr virus (EBV) with 100 newly diagnosed cases of squamous cell carcinoma (SCC) of upper aerodigestive tract (UADT) and correlate its association with clinical parameters, histomorphological grade and staging using immunohistochemical markers p16, LMP1, p53, p63. The study was conducted in the department of Pathology, Maulana Azad medical college and associated hospitals, New Delhi from September 2018-April 2019. Specimens was routinely processed. Immunohistochemistry (IHC) was done using p16, LMP1, p53, p63 monoclonal antibodies. SPSS 20 software. The quantitative analysis was done using Pearson chi-square test. Probability values < 0.05 was be considered statistically significant. HPV was present in 29% cases while EBV in 38% cases. Oral cavity was the most common site involved by both HPV and EBV. Co-infectivity was found in 4% cases. There was significant male predominance in both. HPV was more prevalent in age >45 years while EBV was equally distributed in <45 years and >45 years. Moderately differentiated squamous cell carcinoma was the commonest grade involved by both the viruses. A significant correlation was found between EBV and alcohol. p53 positivity had an inverse relationship with HPV positivity. P63 expression was higher in HPV and EBV positive cases. In resource constraint settings, p16 and Latent membrane protein 1 can be used as surrogacy markers for Human Papilloma virus and Epstein Barr virus along with p53 and p63 for its association with histomorphological grade and stage.

The role of p63 Myoepithelial cell marker in papillary lesions of the breast

Background: To diagnose various histological types of papillary lesions of the breast according to the newer WHO classification and to study the expression of myoepithelial cell marker p63 in various papillary lesions. Material and Methods: The study was done on 21 patients with breast lesions between Jan 2015 to June 2016. Breast biopsies (incisional as well as excisional) were retrieved from the histopathology section of the department. Result: P63 showed a nuclear positivity in myoepithelial cells. It was retained in the benign papillary lesions with some exceptions and was almost completely lost in malignant papillary neoplasms Conclusion: P63 can be used to observe the characteristic pattern of nuclear staining and delineate the myoepithelial layer wherever needed.

Diagnostic Accuracy of FNAC and Ultrasonography in Salivary Gland Lesions in Comparison with Histopathology

Introduction: Modern era demands for early and accurate diagnosis. Fine Needle Aspiration Cytology (FNAC) and Ultrasonography (USG) are two such diagnostic modalities which have gained importance in recent years due to their rapid, repeatable and precision properties. Aim: To study the accuracy of FNAC and USG in diagnosing salivary gland lesions in comparison with histopathology and to study the expression of p63 and Cytokeratin 7 (CK7) Immunohistochemical (IHC) markers in malignant salivary gland tumours. Materials and Methods: A cross-sectional study was conducted on 106 cases who presented with salivary gland lesions possessing USG reports and underwent FNAC and surgical specimens received in Cytopathology and Histopathology Department, Government Medical College Thiruvananthapuram, Kerala, India during one year period (February 2017-January 2018). In each case FNAC was performed and slides were stained with Papanicolaou and Giemsa, surgical specimens were grossed, histopathological features studied under microscope and tumours classified according to World Health Organisation (WHO) Classification of Salivary gland Tumours 2017. Diagnosis on USG and FNAC were compared with histopathological diagnosis. Sensitivity, specificity, Positive Predictive Value (PPV), Negative Predictive Value (NPV), diagnostic accuracy, concordance and discordance was calculated by Statistical Package for the Social Sciences (SPSS) software 24.0 version. p63 and CK7 IHC markers were done only in malignant salivary gland tumours. Results: In the present study majority of lesions i.e. 82 cases (77.4%) were seen predominantly in parotid gland and pleomorphic adenoma was the most commonest lesion and neoplasm overall with 50 cases (47.2%). Most common malignant neoplasm was mucoepidermoid carcinoma (four cases, 3.8%). USG and FNAC had high diagnostic accuracy of 89.6% and 97.2% in comparison to histopathology in diagnosing salivary gland lesions. Conclusion: High sensitivity and specificity of USG and FNAC makes them most acclaimed preoperative diagnostic modality for salivary gland lesions, but histopathology remains gold standard. Subtyping of malignant and cystic lesions were difficult in USG and cytology

Audit of prostatic diseases diagnosed from Tru-Cut biopsies in Nnamdi Azikiwe University Teaching Hospital, Nnewi, using the P63 immunohistochemical marker

Background: There is growing literature on the burden of prostatic lesions: both benign and malignant among men of West African ancestry. There are also some studies on the prostate in Anambra State and South-East Nigeria, but these are mainly on patterns and prevalence, without the use of immunohistochemistry. Aims and Objectives: The aim of this study is to determine the histological pattern of prostatic diseases diagnosed from Tru-Cut biopsies in Nnamdi Azikiwe University Teaching Hospital (NAUTH), Nnewi, South-East Nigeria, using the p63 immunohistochemical marker. This will be a pioneer study in both the state and the geopolitical zone. Materials and Methods: This is a 2-year (January 2019 to December 2020) retrospective study of the immunohistochemical expression of p63 on the haematoxylin and eosin-diagnosed Tru-Cut biopsies of the prostate in the archives of the histopathology laboratory in NAUTH, Nnewi. A Purposive sampling method was used with a sample size of 151. Sections were made from the blocks, reviewed and stained using the p63 immunohistochemical marker. Results: There were 191 cases of prostate diseases histologically diagnosed, of which 151 met the inclusion criteria for the study. Of the 151 cases, 97 (64.2%) were diagnosed as benign and 54 (35.8%) were diagnosed as malignant. Following p63 immunohistochemistry, 97 cases were accurately diagnosed as benign and 54 cases accurately diagnosed as malignant with no misdiagnoses. The age of the patients ranged from 50 years to 90 years; however, the predominant population was in the sixth decade (31.8%) and seventh decade (35.1%), with a mean age of 75.3 years. There is no significant difference noted in the mean age of the non-neoplastic and neoplastic groups. Conclusion: This study shows that morphology alone is sufficient for the diagnosis of both Benign and prostatic lesions, as shown by the accuracy level (100%) in the haematoxylin and Eosin results. This is particularly important in resource-poor settings such as ours.

MiR 206 inhibits reorganization of the cytoskeleton in melanoma cells by targeting DDX5

Purpose: To investigate the role and mechanism of microRNA-206 (miR-206) in cytoskeleton reorganization in melanoma cells.&#x0D; Methods: MiR-206 and RNA helicase p68 (DDX5) expression levels were measured in A375, A875, and HEM-M cells by quantitative real time polymerase chain reaction (qRT-PCR). A DDX5 overexpression cell line was constructed, and DDX5 overexpression, A375, and A875 cells were transfected with miR-206 mimic or DDX5 small interfering RNA (siRNA). Transwell assay was used to assess cell migration and invasion of A375 and A875 cells, while Luciferase reporter assay was used to determine the putative target of miR-206. DDX5, miR-206, vinculin, coronin3, and ezrin expression levels were evaluated by qRT-PCR. Protein expressions of DDX5, vinculin, coronin3, and ezrin were evaluated by western blot analysis.&#x0D; Results: DDX5 expression was higher and miR-206 expression lower in A375 and A875 cells when compared to HEM-M cells (p &lt; 0.05). Knockdown of DDX5 and overexpression of miR-206 repressed invasion and migration, and inhibited expression of vinculin, coronin3, and ezrin in A375 and A875 cells (p &lt; 0.05). However, overexpression of DDX5 reversed the effect of miR-206 on cytoskeletal protein expression. Luciferase reporter assay data confirmed that DDX5 is a direct target of miR-206 (p &lt; 0.05).&#x0D; Conclusion: MiR-206 suppresses reorganization of the cytoskeleton in melanoma cells by targeting DDX5, and is thus, a promising target for the treatment of melanoma.

TAp63 regulates bone remodeling by modulating the expression of TNFRSF11B/Osteoprotegerin

ABSTRACT The transcription factor p53 has been shown to control the differentiation process of the mesenchymal stem cells (MSCs). As a matter of fact, in vivo p53 loss leads to an unbalance between bone formation versus bone erosion. Using as experimental system, human bone marrow-derived MSCs and mouse bone marrow-derived TAp63-/- MSCs, we have asked whether the other members of the p53 family, p63 and p73, are involved in controlling MSCs osteogenic differentiation. Our results indicate that both during human and mouse MSC-induced osteogenic differentiation, TAp63 isoforms are mainly upregulated in comparison with p73 isoforms. In addition, MSCs derived from TAp63 knock-out mice show delayed osteogenic differentiation. Interestingly, we found that, in contrast to p53, TAp63 trascriptionally regulates osteoprotegerin (OPG or TNFRSF11B) important for bone remodeling and osteoclastogenesis inhibition throughout the RANKL/RANK/OPG pathway. Analysis of the expression of p63 and OPG in breast cancer, which is one of the most common human cancers that metastatizes to bone, showed that p63/OPG pathway is deregulated and that a higher expression of both p63 and OPG is associated with a better patient survival, suggesting that p63/OPG axis may be further investigated as clinical biomarker for breast cancer metastasis. Abbreviations MSC, mesenchymal stem cells; OPG, osteoprotegerin; RUNX2, Run-trelated transcription factor 2

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.

Clinicopathological characteristics of SMARCB1(INI1)-deficient sinonasal carcinoma

Objective: To investigate the clinicopathological characteristics, diagnosis, differential diagnosis and prognostic factors of SMARCB1 (INI1)-deficient sinonasal carcinoma (SDSC). Methods: Sixteen cases of SDSC diagnosed in the Department of Pathology, Beijing Tongren Hospital from January 2016 to September 2020 were enrolled. Ninety-nine cases of small round cell malignant tumors of the head and neck were selected as the control, including poorly-differentiated squamous cell carcinoma (n=10), poorly-differentiated adenocarcinoma (n=5), undifferentiated carcinoma (SNUC, n=4), NUT carcinoma (n=5), neuroendocrine carcinoma (n=10), and other non-epithelial tumors [olfactory neuroblastoma (n=10), rhabdomyosarcoma (n=10), NK/T-cell lymphoma (n=10), malignant melanoma (n=10), Ewing's sarcoma/primitive neuroectodermal tumor (EWS/PNET, n=5)] and non-keratinizing undifferentiated nasopharyngeal carcinoma (n=20). The clinical and pathologic characteristics of SDSC, and immunohistochemical (IHC) expression of broad-spectrum CKpan, CK7, CK8/18, CK5/6, p63, p40, p16, INI1, NUT and neuroendocrine markers (Syn, CgA, CD56) were evaluated. In situ hybridization (ISH) was used to detect EBER and fluorescence in situ hybridization (FISH) to detect INI1 gene deletion. Results: The 16 cases of SDSC accounted for 1.3% (16/1 218) of all malignant sinonasal tumors in the author's unit during this time period, and 2.4% (16/657) of all malignant epithelial tumors. Microscopically, there was no clear squamous and adenomatous differentiation, but "rhabdoid-like" cells, are often seen. All SDSC cases were positive for CKpan and CK8/18, negative for INI1; Epstein-Barr virus was not detected by ISH; and INI1 gene deletion was observed in all 11 SDSC patients with FISH. Twelve cases were followed up for 3-47 months. One died of tumor-related diseases half a year after diagnosis, and the remaining patients were alive with tumor, the longest survival time was 47 months. Conclusion: SDSC should be differentiated from a variety of poorly-differentiated tumors in the sinonasal area. Histologically, SDSC has no clear differentiation, but the tumor cells are characteristically basal-like or rhabdoid-like, with non-specific vacuoles, translucent or vacuolar nuclei, prominent nucleoli and necrotic foci. They are negative for INI1 IHC staining, and FISH demonstrates INI1 gene deletion. The clinical prognosis is still unclear, further studies on its biologic behavior and treatment methods are warranted.