P53 Expression Research Articles

Proteomic biomarkers profiling in Pakistani pancreatic ductal adenocarcinoma population: a retrospective cohort study.

Aim: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive cancers. Research in various cancers suggests that investigating target biomarkers may provide directions to precision medicine. However, expression of biomarkers varies across different populations. The biomarker profile of Pakistani patients with PDAC remains unexplored.Materials & methods: We conducted a study on 109 patients to analyze a panel of four proteins (KRAS, p53, BRCA1 and APC) using formalin-fixed paraffin-embedded tumor samples. After confirmation of diagnosis and appropriate tumor content, tissues were processed with antibody specific immunohistochemistry experiments. Subsequently, independent microscopic observation was conducted by two pathologists using scoring criteria specific for each antibody.Results: Statistical analysis showed that negative expression of p53 was significantly associated with positive expression of BRCA1 (p=0.000) and APC (p=0.007). The expression of BRCA1 was also found significantly associated with APC (p=0.028). None of the protein showed association with overall survival or patient demographics. Moreover, KRAS expression was shown to be significantly associated with perineural invasion (p=0.005).Conclusion: This is the first study that investigates protein biomarker expression in a large cohort of Pakistani PDAC patients. The findings from the study may provide directions about the population specific biomarkers and targeted therapies for these patients.

Expression of p53 Protein in Different Grades of Gastric Carcinoma

Background: Mutation in the TP53 suppressor gene and accumulation of p53 protein are the common genetic events in gastric carcinomas. This mutation has been associated with abnormalities in cell cycle regulation, DNA repair and synthesis, apoptosis, and suggested to be implicated in the prognosis of gastric carcinoma. Objectives: To assess the immunohistochemical expression of p53 protein in gastric carcinoma and to study the association of p53 protein expression with different histopathological grade of gastric carcinoma. Methods: In this cross-sectional observational study, 60 paraffin blocks of gastric adenocarcinoma were re-evaluated and histopathological grading were assessed. The p53 expression in the tumour cell nucleus of the submitted blocks were assessed by standard immunoperoxidase method. Appropriate positive control was run for each batch of slides. Results: Out of 60 cases, most of them 35 cases were poorly differentiated carcinoma, of which 28(80%) showed high expression and 7(20%) showed low expression of p53 protein. Twenty cases were moderately differentiated carcinoma, of which 12(60%) showed high expression and 8(40%) low expression. Only 5 were well differentiated carcinoma and all of which showed low expression of p53 protein. So, it was seen that with advancing grade of tumor, frequency of high expression of p53 protein was high and yielded statistically significant result (p value = 0.001). Conclusion: There was a significant association between immunohistochemical expression of p53 protein and histopathological grading of gastric adenocarcinoma. These results indicate that immunohistochemical expression of p53 protein is an important prognostic factor of gastric adenocarcinoma, allowing the selection of a group of patients with an extensive therapeutic indication. Mediscope 2024;11(2): 48-51

Hypermethylation of CDKN2A CpG island drives resistance to PRC2 inhibitors in SWI/SNF loss-of-function tumors.

Polycomb repressive complex 2 (PRC2) catalyzes the writing of the tri-methylated histone H3 at Lys27 (H3K27me3) epigenetic marker and suppresses the expression of genes, including tumor suppressors. The function of the complex can be partially antagonized by the SWI/SNF chromatin-remodeling complex. Previous studies have suggested that PRC2 is important for the proliferation of tumors with SWI/SNF loss-of-function mutations. In the present study, we have developed an EED-directed allosteric inhibitor of PRC2 termed BR0063, which exhibits anti-proliferative properties in a subset of solid tumor cell lines harboring mutations of the SWI/SNF subunits, SMARCA4 or ARID1A. Tumor cells sensitive to BR0063 exhibited several distinct phenotypes, including cell senescence, which was mediated by the up-regulation of CDKN2A/p16. Further experiments revealed that the expression of p16 was suppressed in the BR0063-resistant cells via DNA hypermethylation in the CpG island (CGI) promoter region, rather than via PRC2 occupancy. The expression of TET1, which is required for DNA demethylation, was found to be inversely correlated with p16 CGI methylation, and this may serve as a biomarker for the prediction of resistance to PRC2 inhibitors in SWI/SNF LOF tumors.

Design, synthesis, and cytotoxicity screening of novel pyrazolopyrimidines over renal cell carcinoma (UO-31 cells) as p38α inhibitors, and apoptotic cells inducing activities

A series of novel molecules with pyrazolopyrimidine-4-amine core were designed and synthesized as potential cytotoxic agents over Renal Cell Carcinoma cells (UO-31). Results of cytotoxic activity against UO-31 cells showed that pyrazolopyrimidines 19 and 31 were found to be more cytotoxic than sorafenib (SOR). The cytotoxic activity of these compounds appeared to correlate with their ability to inhibit p38α MAPK which are 2.53- and 2.27- folds more potent than SOR. Moreover, results of the cell cycle analysis as well as the results of annexin-V on the (UO-31) cells showed that pyrazolopyrimidines 19 and 31 had a pro-apoptotic activity higher than SOR by 1.42- and 1.20- folds, respectively. Furthermore, compounds 19 and 31 were found to be effective in arresting the cell cycle throughout the accumulation of the cells at G2/M phase. Finally, the tested compounds decreased the TNF concentration as well as increased the expression of tumor suppressor gene p53, Bax/BCL-2 ratio and caspase 3/7.

Assessment of P16 and Ki-67 Proteins Immunoexpressing in Oral Mucosa Among Saudi Smokers.

It is widely known that tobacco use significantly contributes to a variety of health problems, including mouth cancer. The proteins P16 and Ki-67 have a role in regulating the cell cycle and promoting cell growth. Analyzing the levels of these proteins can give us valuable information about the overall health of cells. This study aims to assess the cellular alterations and immunohistochemical expression of P16 and Ki-67 in the oral mucosa of Saudi smokers. From March to December 2023, a cross-sectional study scraped 500 samples from the buccal mucosa. Participants in the study were Saudi citizens of both genders. We selected a total of 300 individuals who smoked cigarettes and 200 individuals who did not smoke tobacco as the control group. The selection process involved two sample techniques: initially purposive sampling and subsequently snowball sampling. The samples underwent an immunohistochemical examination to determine the presence of P16 and Ki-67 protein overexpression. We evaluated the samples by assessing the proportion of cells that exhibited positive staining and the intensity of the staining. The data were examined utilizing SPSS. We identified categorical variables by calculating frequencies and percentages using the chi-squared test. A significance level of p < 0.05 was used to determine statistical significance. A total of 48% of cigarette users had abnormal results, compared to 20% of nonsmokers. These abnormalities included cytological atypia, inflammation, reverse cytological infection, and binucleated or multinucleated cells (p = 0.015). People who smoked had higher levels of P16 and Ki-67 expressions than people who did not smoke, 12% and 5%, respectively (p = 0.042). There were no important changes in P16/Ki-67 expression between participants of different ages (p = 0.68) or between men and women (p = 0.27). These results demonstrate the detrimental effects of smoking on cell health, underscoring the importance of quitting to reduce the risk of cytological abnormalities and related diseases. Smokers have higher levels of the proteins P16 and Ki-67, which shows how important these biomarkers are for understanding the risks to oral health.

Misincorporations of amino acids in p53 in human cells at artificially constructed termination codons in the presence of the aminoglycoside Gentamicin

Readthrough of a translation termination codon is regulated by ribosomal A site recognition and insertion of near-cognate tRNAs. Small molecules exist that mediate incorporation of amino acids at the stop codon and production of full-length, often functional protein but defining the actual amino acid that is incorporated remains a challenging area. Herein, we report on the development a human cell model that can be used to determine whether rules can be developed using mass spectrometry that define the type of amino acid that is placed at a premature termination codon (PTC) during readthrough mediated by an aminoglycoside. The first PTC we analyzed contained the relatively common cancer-associated termination signal at codon 213 in the p53 gene. Despite of identifying a tryptic peptide with the incorporation of an R at codon 213 in the presence of the aminoglycoside, there were no other tryptic peptides detected across codon 213 that could be recovered; hence we constructed a more robust artificial PTC model. P53 expression plasmids were developed that incorporate a string of single synthetic TGA (opal) stop codons at S127P128A129 within the relatively abundant tryptic p53 peptide 121-SVTCTYSPALNK-132. The treatment of cells stably expressing the p53-TGA129 mutation, treated with Gentamicin, followed by immunoprecipitation and trypsinization of p53, resulted in the identification R, W, or C within the tryptic peptide at codon-TGA129; as expected based on the two-base pairing of the respective anticodons in the tRNA to UGA, with R being the most abundant. By contrast, incorporating the amber or ochre premature stop codons, TAA129 or TAG129 resulted in the incorporation of a Y or Q amino acid, again as expected based on the two base pairings to the anticodons, with Q being the most abundant. A reproducible non-canonical readthrough termination codon-skip event at the extreme C-terminus at codon 436 in the SBP-p53 fusion protein was detected which provided a novel assay for non-canonical readthrough at an extreme C-terminal PTC. The incorporation of amino acids at codons 127, 128, or 129 generally result in a p53 protein that is predicted to be ‘unfolded’ or inactive as defined by molecular dynamic simulations presumably because the production of mixed wild-type p53 and mutant oligomers are known to be inactive through dominant negative effects of the mutation. The data highlight the need to not only produce novel small molecules that can readthrough PTCs or C-terminal termination codons, but also the need to design methods to insert the required amino acid at the position that could result in a ‘wild-type’ functional protein.

Effect of Different Concentrations of PRP on the Expression of Factors Involved in the Endometrial Receptivity in the Human Endometrial Cells from RIF Patients Compared to the Controls.

Platelet-rich plasma (PRP) has been suggested for the improvement of endometrial growth and receptivity in the patients with recurrent implantation failure (RIF). The aim of present study was to investigate the impact of different concentration of PRP on the expression of genes involved in the endometrial receptivity in the human endometrial cells from RIF and controls with thin and normal endometrium in vitro. In this cross-sectional study, endometrial biopsies were obtained from 14 healthy fertile women and 14 women with RIF. Endometrial cells from 4 different group (RIF and control with endometrial thickness < 7mm and > 7mm) were cultured with three different concentration of PRP 3%, 5% and 10%. Expression of leukemia inhibitory factor (LIF), COX2 and P53, estrogen receptors (ERs) and progesterone receptors (PRs) genes were measured using quantitative real-time polymerase chain reaction (PCR). Protein expression levels of LIF, COX2 and p53 were evaluated using Western Blot method (WB). There was a significant decrease in the expression of PROA/b, ER2/b, LIF/b, COX2/b and P53/b genes in the RIF groups compared to the controls. Treatment with 5% and 10% PRP caused a significant increase in the gene expression of PRs, ERs, LIF/b, COX2/b and p53 in the RIF groups. Moreover, protein expression of COX2/b, LIF/b and p53/b increased following treatment with PRP in the RIF group with the endometrium thickness < 7mm. PRP enhances expression of LIF, COX2, p53, ERs and PRs in the RIF patients with thin endometrium which may improve endometrium receptivity.

A new bithiophene inhibited amyloid-β accumulation and enhanced cognitive function in the hippocampus of aluminum-induced Alzheimer's disease in adult rats.

Alzheimer's disease (AD) is a progressive and irreversible neurological disorder that gradually deteriorates an individual's ability to carry out even the simplest tasks. This study was undertaken to investigate the potential therapeutic efficacy of a novel bithiophene in a rat model of aluminum-induced AD pathology. A total of 108 adult male albino rats weighing 160 ± 20 g, were randomly assigned to six groups: (1) a control group administered DMSO, (2) group receiving a high dose of bithiophene (1 mg/kg), (3) a model group received AlCl3 (100 mg/kg), those rats were then treated by either (4) bithiophene low dose (0.5 mg/kg), (5) high dose (1 mg/kg), or (6) memantine (20 mg/kg). Low dose bithiophene treatment was a promising strategy for mitigating oxidative stress and improving synaptic plasticity. This was demonstrated by reductions in malondialdehyde level, and increased activities of superoxide dismutase and catalase, and elevated glutathione content. Likewise, low dose bithiophene enhanced synaptic plasticity through a reduction in excitatory glutamate and norepinephrine levels, while increasing dopamine. Moreover, bithiophene significantly downregulated the expression of GSAP, GSK3-β, and p53, which are implicated in AD progression. This treatment also decreased caspase 3 and amyloid-β (Aβ1-42) accumulation in the hippocampus. Finally, behavioral assessments revealed that low dose bithiophene significantly enhanced learning abilities, as proved by Morris water maze. Low dose bithiophene mitigated AD through ameliorating oxidative stress, promoting synaptic plasticity, inhibiting the Aβ accumulation, and enhancing the cognitive functions in a rat model.

Xenotropic and polytropic retrovirus receptor 1 (XPR1) promotes progression of papillary thyroid carcinoma via the BRAF-ERK1/2-P53 signaling pathway.

Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer. Xenotropic and polytropic retrovirus receptor 1 (XPR1), identified as a cellular receptor, plays roles in many pathophysiological processes. However, the underlying function and molecular mechanisms of XPR1 in PTC remain unclear. Therefore, this study aimed to elucidate the role of XPR1 in the process of PTC and the potential mechanisms. RNA-sequencing was performed for gene differential expression analysis in PTC patients' tissues. Immunohistochemical assay, real-time PCR, and western blotting assay were used to determine the expression of XPR1, BRAF, and P53 in PTC tissues. The function of XPR1 on the progression of PTC was explored using in vitro and in vivo experiments. The molecular mechanism of XPR1 was investigated using gene silencing, ELISA, immunofluorescence, western blotting, and real-time PCR assays. We found that XPR1 was markedly upregulated in PTC tissues compared to adjacent noncancerous tissues, suggesting that high expression of XPR1 could be correlated with poor patient disease-free survival in PTC. In addition, the expression of BRAF and P53 in PTC tissues was substantially higher than in adjacent noncancerous tissues. Silencing of XPR1 reduced the proliferation, migration, and invasion capacities of TPC-1 cells in vitro and effectively inhibited the tumorigenecity of PTC in vivo. More importantly, silencing of XPR1 in TPC-1 cells significantly decreased the expression of XPR1, BRAF, and P53 both in vitro and in vivo. Interestingly, we demonstrated that XPR1 may positively activate the BRAF-ERK-P53 signaling pathway, further promoting PTC progression. The findings reveal a crucial role of XPR1 in PTC progression and prognosis via the BRAF-ERK1/2-P53 signaling pathway, providing potential therapeutic targets for treating PTC.

The Effect of Circulating Exosomes Obtained from Young and Old Individuals on the Aging related hTERT and P16 Expression in Hematopoietic Stem Cells

Introduction: Functional reduction of telomeres can induce DNA damage response through cell cycle checkpoints and contribute to the senescence of stem cells. The effect of exosomes on the aging and rejuvenation of hematopoietic stem cells (HSCs) is not well known. Therefore, the present study is designed to examine the impact of plasma exosomes derived from young and old individuals on hTERT and P16 expression involved in the cellular aging process. Methods: Exosomes isolated from four young (Y-Exo) and four old (O-Exo) men were evaluated for CD63 protein expression, morphology, size and zeta potential. HSCs were treated with exosomes, and then, the cell viability and the mRNA expression (hTERT and P16) were evaluated using MTT and qRT-PCR methods, respectively. To measure the hTERT protein level, a western blot technique was performed. Results: The gene expression of hTERT was significantly decreased in HSCs treated with 5 μg/ml (O5-Exo) and 10 μg/ml (O10-Exo) doses of exosomes obtained from elderly individuals compared to the cells treated with young exosomes and the untreated HSCs (p &lt; 0.05). In addition, there was a profound elevation of hTERT protein in the HSCs treated with both doses of young exosomes in comparison with the cells treated with both doses of old exosomes (p &lt; 0.05). Moreover, P16 expression was markedly upregulated in the O5-Exo and O10-Exo groups compared to the untreated group (p &lt; 0.05). Conclusion: Our findings reinforce the concept that depending on the age of individuals, circulating exosomes may acquire properties that affect the pathways involved in the aging process in HSCs.

Lateral Flow Assay for Cattle Age Determination: Implications for Diagnosing Bovine Spongiform Encephalopathy

Quarantine authorities in East Asia prohibit the import of cattle over 30 months old due to concerns about bovine spongiform encephalopathy (BSE). Accurately determining the age of cattle is critical for complying with these regulations. However, current methods for determining cattle age are time-consuming and require laboratory analysis, making them impractical for real-time use during import inspections. This creates a pressing need for a point-of-care test (POCT) method that can rapidly determine cattle age from beef samples in the field. 4To develop a portable POCT method capable of quickly and accurately determining the age of cattle from beef samples in the field. We investigated the expression of cell cycle proteins p53 and p21, and based on these findings, designed a lateral flow assay (LFA) to measure cattle age using the expression patterns of these proteins.We investigated the expression of cell cycle proteins p53 and p21 using immunoblot analysis to determine the age of beef and applied these findings to lateral flow assay (LFA). The expression pattern of p53 remains at baseline levels up to 28 months and then increases sharply after 35 months. In contrast, p21 expression gradually decreases, reaching baseline levels after 30 months. The SPR sensogram of p21 using surface plasmon resonance (SPR) clearly distinguished beef samples between 27-month-old and 31-month-old cattle. We constructed a calibration curve using recombinant p21 protein to determine the age of beef samples from cattle aged 9 to 33 months. The LFA sensor strip, designed with bovine p21 epitope-based peptide antibody, improved sensitivity and limit of detection (LOD) by more than 10 times.

HER2 status, MMR, and p53 expression among endometrial cancer patients

HER2 status, MMR, and p53 expression among endometrial cancer patients

BCL11A expression worsens the prognosis of DLBCL and its co-expression with C-MYC predicts poor survival

Non-Hodgkin's lymphoma (NHL) is a significant global malignancy, with diffuse large B cell lymphoma (DLBCL) being the most prevalent subtype, accounting for 25-50% of newly diagnosed cases in China. Despite a 60% survival rate achieved with R-CHOP regiment for DLBCL, approximately 40% of patients experience relapse or develop resistance to treatment. While the oncogenic transcription factor B-cell chronic lymphocytic leukaemia/lymphoma 11A (BCL11A) has been implicated in various tumors, its specific role in DLBCL remains unclear. In this study, we conducted retrospective histomorphological and immunophenotypic analyses on paraffin sample tissues and collected fresh tissue samples for protein and mRNA analyses to investigate the relationship between BCL11A and DLBCL. Additionally, we classified DLBCL into subtypes based on cells of origin (COO) and examined the expressions of BCL11A, C-MYC, P53 and other protein expressions to better understand the factors contributing to poor clinical outcomes in DLBCL. Our findings revealed elevated BCL11A expression in DLBCL, with increased expression associated with worse prognosis and higher C-MYC expression. Patients exhibiting co-expression of C-MYC and BCL11A had significantly lower survival rates compared to those with singular expression. Furthermore, BCL11A protein expression levels demonstrated significant associations with P53 and C-MYC protein expression levels in the Germinal Center B-cell-like (GCB) subtype. These findings suggest that BCL11A may serve as a potential prognostic marker and therapeutic target for DLBCL.

Efficacy and Mechanisms of Jiyin Fang (Granule) Against Postmenopausal Coronary Heart Disease Complicated with Osteoporosis

Purpose To explore the efficacy and mechanisms of Jiyin Fang (JYF) against postmenopausal coronary heart disease (CHD) complicated with osteoporosis (OP). Methods Firstly, collected ingredients and relative targets of JYF from TCMSP and BATMAN-TCM database, disease targets of CHD and OP from GeneCards, TTD and OMIM database. By use of protein to protein (PPI) network, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, obtained core targets and mechanism pathways. Finally, preliminarily explore the pharmacological effect and major targets partly to explain the mechanism, through experiment of ovariectomized CHD with OP model rats treated with JYF. Results Total of 112 ingredients and 253 related targets of JYF, 173 disease targets common to CHD and OP were screened out. PPI network indicated 9 core targets. GO and KEGG enrichment analysis showed 20 major signal pathway. Rats experiment displayed that JYF could ameliorate the structural injury of aortic arch and femur; reduce the abnormal changes of ST segment and T wave of ECG; decrease the content of BGP, OPG, ALP, CTX, IL-6 and TNFα, increase the levels of PINP in serum; inhibit P53, MAPK, NF-κB and IKK protein expression, promote PPARG protein expression of aortic arch; inhibit NRF2, AKT protein expression and up-regulate p-STAT1, MMP9 protein expression in femur. Conclusion JYF can achieve the purpose of anti postmenopausal CHD complicated with OP. The potential mechanism may relate to P53/MAPK/NF-κB in aortic arch, and AKT/NRF2/p-STAT1 in bone, due to IL-6 and TNFα.

Comprehensive Clinicopathological and Immunohistochemical Analysis of Human Papillomavirus-independent Squamous Cell Carcinoma and Adenosquamous Carcinoma of the Uterine Cervix.

Human papillomavirus-independent (HPVI) squamous cell carcinoma (SCC) and adenosquamous carcinoma (ASC) of the uterine cervix are extremely rare. The aim of this study was to comprehensively describe the clinicopathological features, patient outcomes, and immunophenotypes of HPVI SCC and ASC. We found four and two patients with HPVI SCC and ASC, respectively, and reviewed their electronic medical records and pathology slides. We also performed immunostaining for p16 and p53. All except one patient underwent surgery. Two, one, and one patients with HPVI SCC were diagnosed as having IIIC1, IVA, and IVB diseases, respectively. Two patients with HPVI SCC experienced recurrences, and died of disease within nine months after treatment initiation. Both patients with HPVI ASC developed lung metastasis at four months post-operatively. HPVI SCCs and the squamous component of HPVI ASCs showed keratinizing, condylomatous, or poorly differentiated morphology. The glandular component of HPVI ASCs was gastric-type endocervical adenocarcinoma. None of the six tumors exhibited block positivity for p16. Two HPVI SCCs and one HPVI ASC displayed aberrant p53 expression. HPVI SCC is a rare and aggressive cervical malignancy that presents initially as advanced-stage disease with poor prognosis. Although the patients with initial stage I and II HPVI ASC were treated with curative intent, distant metastases appeared in the lungs during the early course of treatment. Further investigations are necessary to clarify the association between histological features and clinical behavior of HPVI ASC.

044. Unlocking The Potential: Comparing The Effects of Probiotics Alone and in Combination with Capecitabine Against Colorectal Cancer in Rats

Background: Colorectal cancer (CRC) is a leading cause of cancer deaths globally. While capecitabine is an effective chemotherapy for CRC, its use is limited by side effects and resistance. This has sparked interest in combining capecitabine with natural therapies, such as probiotics from dadiah, a traditional West Sumatra fermented buffalo milk. Lactococcus lactis strains D4 in dadiah produce nisin and butyrate, both known for their anti-cancer properties. To compare the effects of Lactococcus lactis D4, capecitabine, and their combination on CRC progression in a rat model induced with 12-Dimethylhydrazine (DMH). Methods: A total of 25 male Sprague-Dawley rats were divided into five groups: negative control, CRC-induced positive control, probiotics, capecitabine, and combination. After CRC induction, the rats received treatment for 14 days. Nuclear factor kappa B (NFKB) (inflammation), tumor protein 53 (P53) (apoptosis), and Cyclin D (proliferation) expressions were measured using immunohistochemistry. Results: The combination group showed the greatest reduction in NFKB expression compared probiotics-only and capecitabine-only (12.99 ± 4.91 vs 24.15 ± 5.50 vs 16.07 ± 3.79) (p &lt; 0.001); P53 expression increased in the probiotic and capecitabine groups but decreased in the combination group (3.60 ± 3.50 vs 4.40 ± 4.27 vs 3.00 ± 2.00); and Cyclin D was lower in the combination group compared both the probiotic and capecitabine groups (11.68 ± 4.78 vs 21.98 ± 6.05 vs 12.16 ± 4.65). Conclusion: Lactococcus lactis D4 combined with capecitabine synergistically reduces inflammation and cell proliferation in CRC, indicating potential as an adjunct therapy to enhance chemotherapy efficacy.

Vulvar squamous cell carcinoma: The role of p53 and p16 immunohistochemistry

Vulvar squamous cell carcinomas (SCC) represent a heterogeneous group of patients with implications for prognosis and response to treatment. Human papillomavirus (HPV)-associated SCC is characterised by p16 positivity, whereas non-HPV SCC often shows aberrant p53 expression. We conducted a retrospective analysis involving 148 patients with vulvar SCC from two Gynecologic Oncology units from Sydney, Australia. Patients’ demographics, tumor characteristics, types of treatment and survival were analyzed and compared to p16 and p53 immunohistochemistry status. The p16-positive group was younger and included a higher prevalence of smokers, while the p53-positive group demonstrated greater comorbidity indices and was associated with tumor features that are independently related to poor prognosis. Compared to p16-positive patients our study has shown significantly higher recurrence rates and lower overall survival in the p53-positive group. Our findings support existing literature, emphasizing the prognostic significance of p16 and p53 in vulvar SCC. Despite the retrospective nature and variations in immunohistochemistry reporting, our study provides valuable insights into patient outcomes, particularly in a demographically diverse population. Future research, like the STRIVE trial, may determine if implementation of p16 and p53 stratified management algorithm will improve outcomes for women with vulvar SCC (McAlpine, 2024).

EXPRESSION OF p16 AND Ki-67 IN ORAL SQUAMOUS CELL CARCINOMA

Introduction:Oral Squamous Cell Carcinomais a major health challenge with high recurrence and poor prognosis.Asper Globocan 2022, lip and oral cancer ranked 16th in terms of incidence with 389,485(2%)newcases. p16 helps identify HPV infection and induces apoptosis, while Ki-67, a key cell cycle regulator, is crucial for assessing cell proliferation, tumor development and prognosis. Aim: The study aims to evaluate the immunohistochemical expression of p16 and Ki-67 in oral squamous cell carcinoma and correlation with various clinicopathological parameters. Material and Methods: This descriptive analysis was conducted at Muzaffarnagar Medical College &amp; Hospital, involving 50 histologically confirmed oral squamous cell carcinoma cases. The study involved p16 and Ki-67 expression through immunohistochemistry on histologically diagnosed OSCC cases and scoring was done. Results: The study found that most OSCC patients belong to fifth and sixth decades of life, with buccal mucosa being the most affected site. Tobacco use was prevalent among the patients. Most cases were moderately differentiated. p16 and Ki-67 expression werehigher in poorly differentiated cases. Positive p16 expression was observed in 38% of the cases but showed no significant correlation with age, gender, or histological grade. However, significant associations were found between age, histological grade, and Ki-67 expression. Conclusion: This study concluded that p16 expression was positive in 38% of OSCC cases, with higher rates in poorly differentiated tumors, but no significant links to age, gender, or histological grade. Ki-67 levels, higher in poorly differentiated cases, were significantly associated with age, indicating more aggressive disease in older ones.

Electrospun nanofibrous scaffolds reinforced with therapeutic lithium/manganese-doped calcium phosphates: Advancing skin cancer therapy through apoptosis induction

In the current study we fabricated potent materials by incorporating therapeutic elements into calcium phosphates (CPs) to combat cancer. This involved synthesizing manganese (Mn)- and lithium (Li)-doped CPs and loading them into electrospun nanofibers (NFs) composed of chitosan (CS) and polyethylene oxide (PEO). The characterized CPs exhibited excellent properties, including a particle size of 47–75 nm, surface charge of –(30−56) mV, and specific surface area of 75–266 m2/g. The electrochemical analysis revealed that Mn and Mn/Li-doped CPs are promising for generating oxygen free radicals and H2O2, crucial for cancer therapy. Biological evaluation showcased the outstanding performance of the developed materials. MTT assay revealed a cytotoxic effect of nano-constructs on melanoma A375 cell line without adverse effects on normal L929 cells over 72 h. Annexin V/PI apoptosis assay indicated substantial apoptosis rates in A375 cells treated with PC-20 % (62.55 ± 4.59 %). The obtained data of qPCR analysis of pro-apoptotic and anti-apoptotic genes (P53, Bax, Bcl-2) in A375 cells treated with different CP nanoparticles (NPs) showed a significant increase in P53 and Bax gene expression, indicating high levels of A375 cell apoptosis. Additionally, the samples containing Mn ion exhibited high reactive oxygen species (ROS) generation. In conclusion, the fabricated NFs scaffolds hold promising potential for cancer therapy.

Electric Stimulation at 448 kHz Modulates Proliferation and Differentiation of Follicle Dermal Papilla Cells

Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Alopecia is a pathology caused by a deregulation in the hair cycle phases. Currently, the use of physical therapies such as radiofrequency (RF) as an alternative to pharmacological treatment is increasing. Electrical stimulation by capacitive resistive electrical transfer (CRET) is one of these therapies. The objective of the present study was to analyze the effect of RF-CRET currents on DPCs. Cells were treated with subthermal 448 kHz CRET currents with two different types of signals: standard (CRET-STD) or modulated (CRET-MOD). Viability (XTT Assay), proliferation (Ki67 and ERK1/2), apoptosis (p53 and caspase 3), differentiation (β-catenin and α-SMA), and anagen markers (versican and PPARγ) were analyzed by immunofluorescence and immunoblot. CRET caused effects on the proliferation and survival of DPCs associated with increases in the expression of p-MAPK-ERK1/2, cyclin D1, and decreases in the expression of p53 and caspase 3. Also, CRET caused significant transient increases in the expression of β-catenin, involved in hair growth, and in the expression of anagen phase markers such as versican and PPARγ related to hair follicle maintenance. The present study highlights the ability of treatment with CRET therapy to cause molecular alterations in DPC involved in hair regeneration.