Ki-67 Protein Research Articles

Deficiency of Pdcd10 causes urothelium hypertrophy and vesicle trafficking defects in ureter.

The scaffolding protein programmed cell death protein 10 (Pdcd10) has been demonstrated to play a critical role in renal epithelial cell homeostasis and function by maintaining appropriate water reabsorption in collecting ducts. Both ureter and kidney collecting duct systems are derived from the ureter bud during development. Here, we report that cadherin-16 (Cdh16)-cre drives gene recombination with high specificity in the ureter, but not the bladder, urothelium. The consequences of Pdcd10 deletion on the stratified ureter urothelium were investigated using an integrated approach including messenger RNA (mRNA) expression analysis, immunocytochemistry, and high-resolution confocal and electron microscopy. Loss of Pdcd10 in the ureter urothelium resulted in increased expression of uroplakins (Upks) and keratins (Krts), as well as hypertrophy of the ureter urothelium with an associated increase in the number of proliferation marker protein Ki-67 (Ki67)-expressing cells specifically within the basal urothelium layer. Ultrastructural analysis documented significant modification of the intracellular membrane system, including intracellular vesicle genesis and transport along the basal- to umbrella-cell-layer axis. Additionally, Pdcd10 loss resulted in swelling of Golgi compartments, disruption of mitochondrial cristae structure, and increased lysosomal fusion. Lack of Pdcd10 also resulted in decreased fusiform vesicle formation in umbrella cells, increased secretion of exosome vesicles, and alteration in microvillar structure on apical membranes. Our findings indicate that Pdcd10 expression and its influence on homeostasis is associated with modulation of endomembrane trafficking and organelle biogenesis in the ureter urothelium.

The effects of the LaSota strain of oncolytic Newcastle disease virus vaccine on cervical intraepithelial neoplasia Patients-Clinical cohort study

BackgroundCervical cancer is one of the most common malignancies in women, and its treatment has many side effects. Therefore, in this research, the effects of the LaSota strain of oncolytic Newcastle disease virus vaccine on cervical intraepithelial neoplasia (CIN) patients were investigated. Methods15 patients who met the inclusion criteria and diagnosed as CIN II and CIN III were included in the study. The vaccine was injected inside the cervix (neoplasia site) at increasing doses during 21 days, and they were evaluated for adverse events. NDV antibody titer was measured in 90 days and the levels of ki-67 and p16 proteins were studied by immunohistochemistry. Also, the levels of some important inflammatory cytokines in the serum of CIN patients were measured and finally the patients were evaluated according to the final outcomes and the reduction of tumor lesions. ResultsOnly in the first dose of vaccine some patients showed flu-like symptoms. The accumulation of NDV antibodies started on the 7th day of the study and increased until the 90th day. Administration of LaSota vaccine had no significant effect on the expressions of Ki-67 and p16 proteins. Nevertheless, a decrease in the serum levels of Il-1β was observed in patients after the administration of the vaccine, but the serum levels of both Il-2 and INF-γ upregulated significantly. Also, vaccine administration had no significant effect in reducing CIN grades and lesions. ConclusionsIn general, we concluded that LaSota strain of NDV vaccine has no therapeutic effectiveness in CIN patients.

Abstract C022: Clinical, morphological, and immunohistochemical characterization of Nigerian prostatic adenocarcinomas: Current challenges and promising global initiatives

Abstract (A) AN INTRODUCTORY SENTENCE INDICATING THE PURPOSES OF THE STUDY: Prostate cancer, a major leading cancer among men worldwide, still poses a considerable medical and public health challenge in many parts of the world. Despite advances in screening and multimodal management of this disease, overall survival remains poor. The need to identify tumour markers as indicators for prognosis and targets for new therapeutic strategies, still remains a major challenge in prostate carcinoma research. The present study was conducted at a tertiary care teaching hospital at Uyo, South-South Nigeria to assess the expression of Ki-67 and p53 in histologically proven cases of prostatic carcinoma and to evaluate any correlation between the two as well as correlating the expression of both markers with tumour grading and scoring. (B) A BRIEF DESCRIPTION OF PERTINENT EXPERIMENTAL PROCEDURES: The study was conducted on120 histopathologically proven cases of prostate carcinoma received in ourTeaching Hospital, Uyo, South South Nigeria. Formalin fixed, paraffin embedded tissue blocks of prostate adenocarcinoma of those patients with complete clinical details were retrieved, stained and analysed for p53 and Ki67 proteins using immunohistochemistry. The results were analyzed. (C) A SUMMARY OF THE NEW, UNPUBLISHED DATA: The mean age of the prostate cancer patients was 61.7 years + 4.5 (range: 45–96 years). The most common pattern of Gleason grade seen was Group 5 (5+4) constituting 46.0% of the total cases while the Gleason score of 8-9 was the most predominant, constituting 65.0% of the total cases. The p53 positivity was observed in 31.0% of cases with percentage positive cells varying from 3-44% with mild, moderate and strong staining intensity. It was observed that with increase in the grade and score the number of cases showing positivity also increased with statistical significance seen with the same (p<0.05). The Ki-67 positivity was also observed in 10.0% of cases with percentage positive cells varying from 2-24% with moderate and strong staining intensity. It was observed that with increase in the grade and score the number of cases showing positivity also increased but no statistical significance was seen with the same. Five cases (3.3%) were positive for both p53 and Ki-67. Of the 5 cases of prostatic carcinoma positive for both Ki-67 and p53, 4 were seen in high grade tumours (80.0%) and there was statistical significance observed with increase in grade and score ((p<0.05).(D) CONCLUSIONS: The present study highlighted that the immunohistochemical expression of Ki-67 and p53 in prostate carcinoma was lower when compared to other studies. They also permit development of prognostic factors as their expression increases with increase in the grade and score as well as offering of appropriate targeted treatment leading to increase in the survival time. It is hoped that research in unraveling biology of prostatic adenocarcinomas among patients of African Ancestry (AA) and Caucasian Ancestry (CA) using more biomarkers will be carried out in the future. Citation Format: Emmanuel Kunle Abudu, Agility Ukyi-Osuwa Obi-Ihesie, Ikwo Jonathan Kudamnya, Oluwasayo Omolara Abudu. Clinical, morphological, and immunohistochemical characterization of Nigerian prostatic adenocarcinomas: Current challenges and promising global initiatives [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C022.

Abstract C021: Clinicopathological features and basal-like phenotype expression in triple negative breast cancer at Uyo, Niger-Delta region of Nigeria

Abstract (A) AN INTRODUCTORY SENTENCE INDICATING THE PURPOSES OF THE STUDY: Patients of African Ancestry (AA) experience higher rates of TNBC and mortality than European Ancestry (EA) patients, despite lower overall breast cancer (BC) incidence. BC is the most frequently diagnosed cancer in Nigeria and USA as well as being the second most common cause of cancer-related death in women, although there have been significant advances in treatment strategies over the past few decades. In addition to conventional therapies, in the molecular era, breast cancer patients often receive targeted treatments based on their unique cancer biology in both adjuvant and neoadjuvant settings. There is limited data on the prevalence of Basal-like breast carcinomas in Nigeria owing to the high cost of gene expression profiling which is the gold standard for molecular classification and immunohistochemistry (which can be used as a surrogate). This study aimed to analyze the clinico-pathological features and immunohistochemical expression of basal-like phenotype in TNBC at Uyo, as well as stratifying of invasive breast carcinomas into main molecular subtypes and correlating the Basal-like breast carcinoma subtype with the size of tumour, grade and age of patient. (B) A BRIEF DESCRIPTION OF PERTINENT EXPERIMENTAL PROCEDURES: One hundred and fifteen formalin fixed, paraffin embedded blocks of Invasive breast carcinomas were assembled, stained and analysed for ER, PR, HER-2, Ki67, CK5/6 and EGFR proteins using immunohistochemistry. Stratification of invasive breast carcinomas into four (4) main molecular groups (Luminal, HER-2, Basal-like and normal breast subtypes) was done. Patient’s record and immunohistochemical results were reviewed. The data was analyzed and their results were presented as tables, graphs and photomicrographs. (C) A SUMMARY OF THE NEW, UNPUBLISHED DATA: The mean age of the TNBC patients was 39.0 years (range: 25–74 years). Invasive carcinoma (no special type) was the predominant histologic subtype (90.4%). Out of the 115 cases, ER, PR and HER-2 were positive in 53.0 %, 33.0 % and 12.2 % of cases respectively. CK5/6 and EGFR were also positive in 16.5% and 11.3 % of cases respectively. Ki67 was high (>14 %) in 26.1 % of cases. Our study illustrated that majority of TNBC expressed basal-like markers (51.3 %) and occurred at the 35-44 years age group (33.3%) with a substantial proportion being grade 3 (poorly differentiated) carcinomas (48.7%). There was a significant association between basal-like breast carcinoma and tumour grade.(D) CONCLUSION: Our study illustrated that majority of TNBC expressed basal - like markers and occurred at 35-44 years age group with a substantial proportion being grade 3 (poorly differentiated) carcinomas. There was a positive correlation between basal-like breast carcinoma and tumour grade. It is hoped that research in unraveling biology of basal-like carcinomas among patients of African Ancestry (AA) and Caucasian Ancestry (CA) will be carried out in the future. Citation Format: Emmanuel Kunle Abudu, Obioha Kanu, Ikwo Jonathan Kudamnya, Emem Imo Akpanudo, Oluwasayo Omolara Abudu. Clinicopathological features and basal-like phenotype expression in triple negative breast cancer at Uyo, Niger-Delta region of Nigeria [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C021.

Effect of Lysine Acridone Acetate on the Level of Apoptosis and Expression of Apoptosis-Associated Proteins during Antioncogenic Therapy in Colorectal Cancer in Mice.

We studied the mechanism of action of cytostatics with the addition of lysine acridone acetate to evaluate the possibility of its use for improving the effectiveness of antioncogenic therapy in colorectal cancer. In Nude mouse model, the level of apoptosis (TUNEL) and expression of proteins CD95, p53, Bcl-2, histone H3, and Ki-67 (immunohistochemistry) were assessed in primary tumor biopsy specimens. It has been shown that cytostatic treatment led to stimulation of p53-mediated apoptosis and suppression of proliferation (Ki-67 expression) of tumor cells, and apoptosis level was increased in groups receiving lysine acridone acetate. H3 expression in the experimental groups was changed.

Correlation between the expression of p16/INK4a and Ki-67 proteins and the histopathological diagnostic in women infected by Human papillomavirus in the Agreste region of Pernambuco State, Brazil

Epidemiological studies have stated that cervical infection with high-risk human papillomavirus (HR-HPV) is a necessary but not sufficient cause in the development of cervical cancer. Cervical cancer is preceded by precursor lesions called cervical intraepithelial neoplasia (CIN), which are classified as CIN 1 (cervical intraepithelial neoplasia grade 1), CIN 2 (cervical intraepithelial neoplasia grade 2) and CIN 3 (cervical intraepithelial neoplasia grade 3). Markers such as p16/INK4a and Ki-67 have been studied with the aim of identifying lesions with a high risk of evolution. The objective of this study was to correlate the expression of the biomarkers p16INK4A and Ki-67 with histological diagnosis CIN 1, CIN 2, CIN 3 and cancer in HPV-infected cervical samples.The study involved conducting an analysis of 211 cases of women with histological confirmation to CIN 1, CIN 2, CIN 3 and cervical cancer. The immunohistochemical staining for the p16/INK4a and Ki-67 proteins was carried out through samples taken from the biopsies and waterproof containers that had representative tissues of the tumor. The expression of p16/INK4A was observed in 100% of the cases of cancer, 97.6% of the cases of NIC 3, 83.3% of the cases of NIC 2, as well as in 55.4% of the cases of NIC 1. In addition, the expression of Ki-67 was observed in 100% of the cases of cancer, 98.8% of the cases of NIC 3, 83.3% of the cases of NIC 2, as well as in 69.6% of the cases of NIC 1. The results obtained in this study make it possible to relate the expression of p16/INK4A and Ki-67 to the atypical histological findings found in the samples of patients infected with HPV.

Patient Characteristics Associated with Growth of Patient-Derived Tumor Implants in Mice (Patient-Derived Xenografts).

Background: patient-derived xenografts (PDXs) have defined the field of translational cancer research in recent years, becoming one of the most-used tools in early drug development. The process of establishing cancer models in mice has turned out to be challenging, since little research focuses on evaluating which factors impact engraftment success. We sought to determine the clinical, pathological, or molecular factors which may predict better engraftment rates in PDXs. Methods: between March 2017 and January 2021, tumor samples obtained from patients with primary or metastatic cancer were implanted into athymic nude mice. A full comprehensive evaluation of baseline factors associated with the patients and patients' tumors was performed, with the goal of potentially identifying predictive markers of engraftment. We focused on clinical (patient factors) pathological (patients' tumor samples) and molecular (patients' tumor samples) characteristics, analyzed either by immunohistochemistry (IHC) or next-generation sequencing (NGS), which were associated with the likelihood of final engraftment, as well as with tumor growth rates in xenografts. Results: a total of 585 tumor samples were collected and implanted. Twenty-one failed to engraft, due to lack of malignant cells. Of 564 tumor-positive samples, 187 (33.2%) grew at time of analysis. The study was able to find correlation and predictive value for engraftment for the following: the use of systemic antibiotics by the patient within 2 weeks of sampling (38.1% (72/189) antibiotics- group vs. 30.7% (115/375) no-antibiotics) (p = 0.048), and the administration of systemic steroids to the patients within 2 weeks of sampling (41.5% (34/48) steroids vs. 31.7% (153/329), no-steroids) (p = 0.049). Regarding patient's baseline tests, we found certain markers could help predict final engraftment success: for lactate dehydrogenase (LDH) levels, 34.1% (140/411) of tumors derived from patients with baseline blood LDH levels above the upper limit of normality (ULN) achieved growth, against 30.7% (47/153) with normal LDH (p = 0.047). Histological tumor characteristics, such as grade of differentiation, were also correlated. Grade 1: 25.4% (47/187), grade 2: 34.8% (65/187) and grade 3: 40.1% (75/187) tumors achieved successful growth (p = 0.043), suggesting the higher the grade, the higher the likelihood of success. Similarly, higher ki67 levels were also correlated with better engraftment rates: low (Ki67 < 15%): 8.9% (9/45) achieved growth vs. high (Ki67 ≥ 15%): 31% (35/113) (p: 0.002). Other markers of aggressiveness such as the presence of lymphovascular invasion in tumor sample of origin was also predictive: 42.2% (97/230) with lymphovascular vs. 26.9% (90/334) of samples with no invasion (p = 0.0001). From the molecular standpoint, mismatch-repair-deficient (MMRd) tumors showed better engraftment rates: 62.1% (18/29) achieved growth vs. 40.8% (75/184) of proficient tumors (p = 0.026). A total of 84 PDX were breast models, among which 57.9% (11/19) ER-negative models grew, vs. 15.4% (10/65) of ER-positive models (p = 0.0001), also consonant with ER-negative tumors being more aggressive. BRAFmut cancers are more likely to achieve engraftment during the development of PDX models. Lastly, tumor growth rates during first passages can help establish a cutoff point for the decision-making process during PDX development, since the higher the tumor grades, the higher the likelihood of success. Conclusions: tumors with higher grade and Ki67 protein expression, lymphovascular and/or perineural invasion, with dMMR and are negative for ER expression have a higher probability of achieving growth in the process of PDX development. The use of steroids and/or antibiotics in the patient prior to sampling can also impact the likelihood of success in PDX development. Lastly, establishing a cutoff point for tumor growth rates could guide the decision-making process during PDX development.

Single-atom Cu-attached MOFs as peroxide-like enzymes to construct dual-mode immunosensors for detection of breast cancer typing in serum

Molecular subtype of breast cancer has defined the treatment direction of breast cancer and guided the selection of drugs. Invasive tests such as tissue biopsy have the potential to promote the metastasis of cancer cells. Therefore, the continuous development of high-performance and low-cost in vitro diagnostic tools is crucial for the diagnosis and treatment of cancer prognosis. Liquid biopsy techniques provide noninvasive, real-time, dynamic, multicomponent, quantitative, and long-term observations at the cellular, genetic, and molecular levels in vivo. A Cu-Zr metal-organic framework(MOF) nanomases with monatomic Cu attachment has been synthesized by covalently attaching single atom Cu to the defect site of zirconia clusters in the UiO-66. The catalytic performance of Cu/UiO-66 as a peroxide-like enzyme was verified by enzyme kinetics calculation. Using o-phenylenediamine as substrate, a colorimetric fluorescence dual-mode immunosensor was constructed by introducing blue fluorescent carbon quantum dots (CDs) based on ultraviolet chromogenic reaction and fluorescence internal filtration effect. By magnetic adsorption separation, the background signal of the sensor is reduced and the sensitivity of the sensor is further improved. In 0.001–1 ng/mL detection range, detection of HER2/ER/PR/Ki-67, the result has obvious linear relationship. Among them, the colorimetric detection limit were 0.367/0.379/0.392/0.388 pg/mL; the ratio fluorescence detection limit can reach 0.3, 0.289, 0.257, 0.33 pg/mL, with high sensitivity. The proteins HER2, ER, PR and Ki-67 in serum related to human breast cancer typing were detected with high sensitivity. Finally, the actual serum samples were tested using sensors. According to the confirmed detection criteria, the classification of serum breast cancer was preliminarily explored with an accuracy of 87.5 %. The sensor has high catalytic performance and excellent real-time detection performance of biomarkers, and have practical application potential in human serum level detection. It provides a new method for the classification of breast cancer in serum.

REST-restrained lncRNA EPB41L4A-AS2 Modulates Laryngeal Squamous Cell Carcinoma Development via Regulating miR-1254/HIPK2 Pathway.

LncRNAs have been corroborated to exert crucial effects in malignancies, including laryngeal squamous cell carcinoma (LSCC). Nevertheless, the role and mechanism of EPB41L4A- AS2 in LSCC are inadequately investigated and warrant further exploration. Relevant database was adopted to analyze the relationship between EPB41L4A-AS2 expression level and tumors. The expressions and relationships of EPB41L4A-AS2, RE-1 silencing transcription factor (REST), miR-1254, and homeodomain interacting protein kinase 2 (HIPK2) in LSCC cells were evaluated by qRT-PCR, Pearson's correlation tests, RNA immunoprecipitation, RNA pull-down assay, chromatin immunoprecipitation, database, and dual-luciferase reporter assay. Following the required transfection, the biological behaviors of LSCC cells were examined using cell function experiments. Meanwhile, the levels of Ki-67 and apoptosis-, and epithelial-mesenchymal transition (EMT) pathway-related proteins were quantified with Western blot. Moreover, xenografts in nude mice were constructed, and the tumor volume and weight were measured. Ki-67 positivity was determined by immunohistochemical staining. EPB41L4A-AS2 and HIPK2 were lower-expressed, yet miR-1254 and REST were higher- expressed in LSCC cells. Pearson's correlation assay results exhibited a positive correlation between HIPK2 and EPB41L4A-AS2 and a negative correlation between HIPK2 and miR-1254. Overexpressed EPB41L4A-AS2 diminished the biological behavior, and repressed the levels of Ki-67 and EMT-related markers in LSCC cells whilst enhancing those of apoptosis-related markers. These aforementioned effects were counteracted by miR-1254 mimic. Moreover, EPB41L4A- AS2 overexpression suppressed the growth of tumors and reduced the positive expression of Ki-67 in nude mice. Besides, miR-1254 aggravated the biological behaviors and elevated the levels of Ki-67 and EMT-related proteins in LSCC cells while reducing the levels of apoptosis-related markers via targeting HIPK2. REST-restrained EPB41L4A-AS2 modulates LSCC development via regulating miR-1254/HIPK2 pathway.

Effects of metronidazole on colorectal cancer occurrence and colorectal cancer liver metastases by regulating Fusobacterium nucleatum in mice.

Colorectal cancer (CRC) represents a leading cause of cancer-related deaths. Metronidazole (MNZ) is exceedingly implicated in CRC. This study explored the roles of MNZ in mouse CRC occurrence and liver metastasis (CRLM). Male BALB/c nude mice were subjected to CRC and CRLM modeling, orally administration with MNZ (1 g/L) 1 week before modeling, and disease activity index (DAI) evaluation. Fresh stool and anal swab samples were collected on the morning of the 28th day after modeling. The relative expression of Fusobacterium nucleatum (F. nucleatum) DNA was assessed by quantitative polymerase chain reaction. After euthanasia, tumor tissues and liver tissues were separated and the tumor volume and weight change were measured. The liver tissues were stained with hematoxylin-eosin to quantitatively analyze the metastatic liver nodules. Malignant tumor biomarker Ki67 protein levels in liver tissues/DNA from stool samples were detected by immunohistochemistry/high-throughput 16S rRNA gene sequencing. Bioinformatics analysis was performed on the raw sequence data to analyze microbial community richness (Chao1 index, ACE index) and microbial community diversity (Shannon index). The DAI and F. nucleatum DNA relative expression in feces and anal swabs of the CRC and CRLM groups were raised and repressed after MNZ intervention. MNZ repressed tumor occurrence and growth in mice to a certain extent, alleviated CRLM malignant degree (reduced liver metastases and Ki67-positive cell density/number), and suppressed CRC liver metastasis by regulating intestinal flora structure, which affected the intestinal characteristic flora of CRC and CRLM mice. MNZ suppressed CRC occurrence and CRLM in mice by regulating intestinal F. nucleatum.

General Clinico-Pathological Characteristics in Glioblastomas in Correlation with p53 and Ki67.

Introduction: A glioblastoma is an intra-axial brain tumour of glial origin that belongs to the category of diffuse gliomas and is the most common malignant neoplasia of the central nervous system. The rate of survival at 5 years, from the moment of diagnosis, is not higher than 10%. Materials and methods: In this retrospective study, fifty-four patients diagnosed with glioblastoma, from the Pathology Department of the County Emergency Clinical Hospital of Târgu Mureș, between 2014 and 2017 were included. We studied the clinico-pathological data (age, gender, location, and laterality) and, respectively, the immunoexpression of p53, Ki67, ATRX, and IDH-1 proteins. Results: We observed a statistically significant association between the laterality of the tumour according to the age groups, with the localization on the right side being more frequent in the age group below 65 years of age, while the involvement of the left hemisphere was more prevalent in those over 65 years. Out of the total 54 cases, 87.04% were found to be primary glioblastomas; more than 70% of the cases were ATRX immunopositive; almost 80% of the glioblastomas studied had wild-type p53 profile; and 35% of the cases were found to have a Ki67 index greater than 20%. A statistically significant association between gender and ATRX mutation was found; female cases were ATRX immunopositive in 92% of the cases. Almost 70% of the cases were both IDH-1 and p53 wild-type, and we observed the presence of both mutations in only 3.7% of the cases. Approximately 83% of primary glioblastomas were ATRX positive, respectively, and all IDH-1 mutant cases were ATRX negative. Conclusions: Glioblastomas still represent a multidisciplinary challenge considering their reserved prognosis. In this study, we described the most common clinico-pathological characteristics and IHC marker expression profiles, highlighting a variety of percentage ranges in primary and secondary glioblastomas. Given the small number of studied cases, further prospective studies on larger cohorts are needed in the future to evaluate the role of these immunohistochemical markers as prognostic factors for survival or recurrence.

Microsatellite instability screening in colorectal carcinoma: immunohistochemical analysis of MMR proteins in correlation with clinicopathological features and Ki-67 protein expression

BackgroundDefects in mismatch repair (MMR) system or microsatellite instability (MSI) and detected in colorectal carcinoma (CRC), both in sporadic and more frequently in hereditary cases. Immunohistochemistry (IHC) is the most frequent method for MMR protein deficiency screening in CRCs. In this study, we aimed to evaluate immunohistochemical expression of MMR and Ki-67 in colorectal carcinoma with clinicopathological features.MethodsIn this study, we evaluated the immunohistochemical expression of MMR proteins including MSH6, MSH2, PMS2 and MLH1 in 50 resection materials with colorectal carcinoma. Their expression is correlated with clinicopathological features of patients together, with Ki-67 protein expression in attempt to screen the most significant predictor of microsatellite instability.ResultsOf the 50 cases of cancer colon, 28% were classified as MSI-H, 20% were MSI-L, and 52% were MSS. The most frequent pattern in MSI-H tumors was concurrent loss of MSH6 and PMS2 proteins. There was a significant correlation between MMR protein expression pattern with tumor size, grade, T-classification and stage (0.015, 0.0515, 0.0162 and 0.0391), respectively. MSI-H tumors were located more frequently in right colon, early TNM stage and poorly differentiated and infrequent distant metastases. There was a significant correlation between Ki-67 high expression and MSI status patterns in their common biological aspects distinct from MSI-negative tumors.ConclusionsMismatch repair defective colorectal carcinoma has characteristics clinicopathological features different from MSS tumors. The role of immunohistochemistry (IHC) for MSI evaluation is the easiest and effective way for evaluation of MMR deficiency in colorectal carcinoma.

Oral lipid nanocomplex of BRD4 PROteolysis TArgeting Chimera and vemurafenib for drug-resistant malignant melanoma

BRAF inhibitors (BRAFi) like vemurafenib (VEM) provide initial regression in mutated melanoma but rapidly develop resistance. Molecular pathways responsible for development of resistance against VEM finally converge towards the activation of oncogenic c-Myc. We identified an epigenetic approach to inhibit the c-Myc expression and resensitize BRAFi-resistant melanoma cells. ARV-825 (ARV) was employed as a BRD4 targeted PROteolysis TArgeting Chimera that selectively degrades the BRD4 to downregulate c-Myc. ARV synergistically enhanced the cytotoxicity of VEM in vitro to overcome its resistance in melanoma. Development of ARV and VEM-loaded lipid nanocomplex (NANOVB) significantly improved their physicochemical properties for oral delivery. Most importantly, oral administration of NANOVB substantially inhibited tumor growth at rate of 41.07 mm3/day in nude athymic mice. NANOVB treatment resulted in prolonged survival with 50% of mice surviving until the experimental endpoint. Histopathological analysis revealed significant tumor necrosis and downregulation of Ki-67 and BRD4 protein in vivo. Promising in vivo antitumor activity and prolonged survival demonstrated by NANOVB signifies its clinical translational potential for BRAFi-resistant melanoma.

THE STUDY OF PROGNOSTIC VALUE OF microRNAs (miR-10b AND -155) AND CDKN2A/P16INK4A IN ORAL SQUAMOUS CELL CARCINOMA.

Oral squamous cell carcinoma (OSCC) is one of the most common types of cancer worldwide. Disco- vering novel prognostic markers for OSCC can improve treatment outcomes by allowing for more effective therapy strategies. To identify the prognostic value of CDKN2A (p16INK4a) and miRNAs involved in its regulation as markers of OSCC. The work is based on the results of the examination and treatment of 70 patients with stage II-IV OSCC. miR-10b, -155, and CDKN2A mRNA expression in tumor samples was ana- lyzed by real-time reverse transcription polymerase chain reaction. The expression of p16INK4a and Ki-67 proteins was determined immunohistochemically. No association of CDKN2A mRNA and p16INK4a protein with Ki-67 expression in tumor tissue and clinical pathological parameters of OSCC patients was found. Most of the p16INK4a-positive cases were characterized by a high Ki-67 expression. We found a strong correlation of the studied miRNAs expression levels with lymph node metastasis (r = 0.56 for miR-10b and r = 0.59 for miR-155). Also, there was no difference in miR-10b and -155 expression between p16INK4a+ and p16INK4a- samples. The association of both miRNAs with lymph node metastases was not affected by p16INK4a status. The results indicate the relationship between miR-10b and -155 and the presence of lymph node metastases in OSCC patients, so these miRNAs can be considered as prognostic markers of the disease.

Nitric oxide synergizes minoxidil delivered by transdermal hyaluronic acid liposomes for multimodal androgenetic-alopecia therapy

Androgenetic alopecia (AGA) is a common clinical condition, affecting over 200 million people globally each year. For decades, Minoxidil (Mi) tincture has been the primary treatment for this disease, but its low utilization rate and significant side effects necessitate new therapeutic strategies. Nitric oxide (NO) is a signaling molecule in various physiological processes, including vasodilation, immune responses, and cell proliferation. Herein, we constructed a hyaluronic acid liposome (HL) complex as a novel transdermal delivery system (HL@Mi/NONOate) for NO and Mi, which displayed promising transdermal and hair-regrowth effects. In-depth mechanistic studies revealed three potential pathways of the synergistic AGA therapy. First, NO promoted capillary dilation and accelerated blood flow, thus achieving efficient penetration of Mi. Due to the structural advantage of liposomes, the residence time of the Mi in the skin was prolonged. Moreover, HL@Mi/NONOate promoted cell proliferation and angiogenesis, and upregulated the expression of regulatory factors involved in follicle stem cell differentiation. In the AGA model, HL@Mi/NONOate down-regulated the expression of inflammatory factors, inhibiting the inflammation of follicle and improving the microenvironment of hair regrowth. Concurrently, HL@Mi/NONOate upregulated the expression of Ki67 and PCNA proteins in follicle tissues, inducing follicle regeneration and development, ultimately achieving the synergistic multimodal AGA therapy.

Cigarette smoke extract induces the senescence of endothelial progenitor cells by upregulating p300.

Endothelial progenitor cells (EPCs) are the main source of endothelial cells. The senescence of EPCs is involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). Cigarette smoke extract (CSE) can directly induce the dysfunction and increased expression of senescence-related markers in EPCs cultured in vitro. Histone acetyltransferase p300 is a transcriptional activator, and its changes can lead to cell senescence. The present study investigated whether CSE can induce the senescence of EPCs by upregulating p300. EPCs were isolated from bone marrow of C57BL/6J mice by density gradient centrifugation. The p300 inhibitor C646 and agonist CTPB were used to interfere with EPCs, cell cycle and apoptosis were detected by flow cytometry, the proportion of senile cells was counted by β-galactosidase staining, the protein expression of p300, H4K12, Cyclin D1, TERT and Ki67 were detected by western blot. Compared with the control group, the cell cycle of CSE group and CTPB group were blocked, the apoptosis rate and early apoptosis rate were increased, the proportion of senile cells counted by β-galactosidase staining was increased, the expression of p300 and H4K12 protein were increased, the expression of Cyclin D1, TERT and Ki67 protein were decreased. C646 could partly alleviate the damages caused by CSE. CSE may promote the apoptosis and senescence of EPCs by upregulating the expression of p300 and H4K12 protein, thus preventing the transition of EPCs from G1 phase to S phase, affecting telomerase synthesis, and reducing EPCs proliferation.

Does a 40% Cut-off Point for Ki-67 Expression Have a Role in Identifying the Development of Distant Metastasis Within 2 Years in Locally Advanced Triple Negative Breast Cancer Patients?

Triple negative breast cancer (TNBC) has a higher proportion of patients with distant recurrence or metastasis. Ki-67 has been suggested as an essential factor in cancer grading and prognostic evaluation, although there is still a debate regarding the Ki-67 cut-off value in TNBC. The aim of this study was to determine the role of Ki-67 expression using a 40% cut-off point as a risk factor for developing distant metastasis within two years in patients with TNBC. This analytical observational study was conducted with a case-control design from January 2021-2022. Subjects were divided into two groups (metastasis within two years or more than two years after diagnosis). Bivariate analysis was conducted using chi-square test and odds ratio (OR) was also analyzed. A total of 66 subjects were included. In patients with metastasized TNBC and a Ki-67 expression of ≥40%, 29 patients (55.8%) had metastasis occurring in ≤2 years and 23 patients (44.2%) had metastasis occurring in >2 years; in patients with metastasized TNBC and a Ki-67 expression of <40%, 4 patients (28.6%) had metastasis occurring in ≤2 years and 10 patients (71.4%) had metastasis occurring in >2 years. Chi-square analysis (p = 0.071) indicated no significant association between patients with Ki-67 expression of ≥40% and <40% with metastasis within 2 years [OR 3.152 (confidence interval: 95% 0.875-11.362)]. Ki-67 protein expression of over 40% in patients with locally-advanced TNBC does not indicate a greater risk of distant metastasis in the first two years after diagnosis.

Astragaloside IV inhibits AOM/DSS-induced colitis-associated tumorigenesis via activation of PPARγ signaling in mice

BackgroundColitis-associated colorectal cancer (CAC) is a severe complication of inflammatory bowel disease (IBD), resulting from long-term inflammation in the intestines. The primary cause of CAC is the imbalance of oxidative metabolism in intestinal cells, triggered by excessive reactive oxygen (ROS) and nitrogen (NO) species production due to prolonged intestinal inflammation. This imbalance leads to genomic instability caused by DNA damage, eventually resulting in the development of intestinal cancer. Previous studies have demonstrated that astragaloside IV is effective in treating dextran sulfate sodium salt (DSS)-induced colitis, but there is currently no relevant research on its efficacy in treating CAC. MethodsTo investigate the effect of astragaloside IV against CAC and the underlying mechanism, C57 mice were treated with (20, 40, 80 mg/kg) astragaloside IV while CAC was induced by intraperitoneal injection of 10 mg/kg azoxymethane (AOM) and ad libitum consumption of 2% dextran sulfate sodium salt (DSS). We re-verified the activating effects of astragaloside IV on PPARγ signaling in IEC-6 cells, which were reversed by GW9662 (the PPARγ inhibitor). ResultsOur results showed that astragaloside IV significantly improved AOM/DSS-induced CAC mice by inhibiting colonic shortening, preventing intestinal mucosal damage, reducing the number of tumors and, the expression of Ki67 protein. In addition, astragaloside IV could activate PPARγ signaling, which not only promoted the expression of Nrf2 and HO-1, restored the level of SOD, CAT and GSH, but also inhibited the expression of iNOS and reduced the production of NO in the intestine and IEC-6 cells. And this effect could be reversed by GW9662 in vitro. Astragaloside IV thus decreased the level of ROS and NO in the intestinal tract of mice, as well as reduced the damage of DNA, and therefore inhibited the occurrence of CAC. ConclusionAstragaloside IV can activate PPARγ signaling in intestinal epithelial cells and reduces DNA damage caused by intestinal inflammation, thereby inhibiting colon tumourigenesis. The novelty of this study is to use PPARγ as the target to inhibit DNA damage to prevent the occurrence of CAC.

Ki-67 Index as an Adjunct Histological Marker for Grading Cartilage Tumors: A Pilot Study

Objectives Distinguishing enchondromas (grade 0) from grade 1 chondrosarcomas, recently classified as atypical cartilaginous tumors (ACT), is challenging due to histologic and radiographic similarity. However, diagnostic ambiguity poses a management dilemma: enchondromas are observed, while ACTs are excised. Protein Ki-67 is a cellular proliferation nuclear marker used as a proxy for aggressiveness in other tumors. This retrospective pilot study aimed to characterize the association between grade and Ki-67 index and evaluate its ability to aid differentiation between critical cartilage tumor grades. Methods Tissue blocks from 26 patients treated for enchondromas or chondrosarcomas within the past ten years were exposed to anti-Ki-67 rabbit monoclonal antibody and hematoxylin counterstain. Counting two high-power fields per sample yielded a Ki-67 index. Results There was a strong positive correlation between tumor grade and the Ki-67 index (r = 0.704, P&lt;0.001). One-way analysis of variance (ANOVA) showed significant differences in the Ki-67 index between grades (P&lt;0.001). Tukey’s post-hoc test demonstrated that enchondroma Ki-67 indexes were not significantly different from ACT Ki-67 indexes (P=0.094). Receiver operator curve cutoff index value of 13.4% differentiated enchondroma vs. ACT with 67.9% sensitivity and 42.9% specificity. Conclusion Ki-67 index is correlated with cartilage tumor grade. Although our study did not show a significant difference in the Ki-67 index between enchondromas from ACTs, this study provided a protocol for future investigations. Study limitations include small sample size and reliance on historical tissue blocks of variable preservation quality.

Emerging Role of Decoy Receptor-2 as a Cancer Risk Predictor in Oral Potentially Malignant Disorders.

The aim of this study was to evaluate the expression of the senescence markers, Decoy Receptor 2 (DcR2) and Differentiated Embryo-Chondrocyte expressed gen 1 (DEC1), in oral potentially malignant disorders (OPMDs) to ascertain their possible association with oral cancer risk. The immunohistochemical analysis of DcR2 and DEC1 expression (along with p16 and Ki67 expression) was carried out in 60 patients with clinically diagnosed oral leukoplakia. Fifteen cases (25%) subsequently developed an invasive carcinoma. Correlations between protein marker expression, histological grade and oral cancer risk were assessed. DcR2, DEC1 and Ki67 protein expressions were found to correlate significantly with increased oral cancer risk, and also with an increased grade of dysplasia. Multivariate analysis demonstrated that DcR2 and Ki67 expression are independent predictors of oral cancer development. Our results evidence for the first time the potential of DcR2 as an early biomarker to assess oral cancer risk in patients with oral leukoplakia (HR = 59.7, p = 0.015), showing a superior predictive value to histology (HR = 4.225, p = 0.08). These findings reveal that the increased expression of DcR2 and DEC1 occurred frequently in OPMDs. In addition, DcR2 expression emerges as a powerful biomarker for oral cancer risk assessment in patients with oral leukoplakia.