Identification of EMT signaling cross-talk and gene regulatory networks by single-cell RNA sequencing.

The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-ß, BMP, Wnt-ß-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequencing on MCF10A cells undergoing EMT by TGF-ß1 stimulation. Our comprehensive analysis revealed that cells progress through EMT at different paces. Using pseudotime clustering reconstruction of gene-expression profiles during EMT, we found sequential and parallel activation of EMT signaling pathways. We also observed various transitional cellular states during EMT. We identified regulatory signaling nodes that drive EMT with the expression of important microRNAs and transcription factors. Using a random circuit perturbation methodology, we demonstrate that the NOTCH signaling pathway acts as a key driver of TGF-ß-induced EMT. Furthermore, we demonstrate that the gene signatures of pseudotime clusters corresponding to the intermediate hybrid EMT state are associated with poor patient outcome. Overall, this study provides insight into context-specific drivers of cancer progression and highlights the complexities of the EMT process.

To Access Knowledge Regarding Organ Donation among Healthcare Workers and Their Willingness toward Organ Donation.

BACKGROUND: In India, critical shortage of organ donations, particularly deceased donations, has led to a dire situation in India, with thousands of patients waiting for transplants and a significant number of them succumbing. One of the reasons for the shortage of organs for transplantation is unawareness and prejudiced information about organ donation. Being direct or indirect stakeholders, the knowledge regarding organ donation among healthcare workers may play an important role in the donation process. AIM: To assess the knowledge regarding cadaver organ donation among healthcare workers and their willingness toward organ donation. MATERIALS AND METHODS: It is a cross-sectional offline self-administered questionnaire-based survey conducted among healthcare professionals at tertiary care teaching institutes. Survey was carried out between the months of August to December 2019. A structured questionnaire was used to assess knowledge and willingness toward organ donation. Statistical analyzed was done by using statistical package for social sciences (SPSS) 20.0. All p-values were considered significant at <0.05. RESULTS: A total of 1,039 healthcare professionals participated in the survey. Out of them, 362 (34.8%) were males and 675 (65%) were females. Average age of the healthcare workers participating in survey was 30.81 years, and age ranged from 18 to 60 years. Awareness regarding corneal donation after brain death was found to be maximum (89.7%) and was comparable to that of kidney (86.6%) and heart (83.7%). Participants were unlearned of donation of lungs, pancreas, hands and unaware of heart valve donation. About 45% respondents considered that age affected the donors. About 40% respondents considered younger patients as ideal recipients, while 18.7% respondents considered waiting list patients as ideal recipients. Doctors had highest willingness (78. 3%) for organ donation, followed by nurses (69.9%) and support staff (59.3%) (p < 0.001). Only 119 (11.5%) participants received organ donation cards as against 68.7% willingness toward organ donation (p < 0.01). CONCLUSION: We have observed fair awareness regarding overall cadaver organ donation concept among healthcare workers. There is a need to improve knowledge of extended age criteria and which organs can be retrieved from deceased donor. Authorities have to work hard on delivery of organ donation pledging card to promote donation program.

Prevalence of suspected Developmental Coordination Disorder and its association with preterm and low birth weight in 5-10-year old children.

Background: Developmental Coordination Disorder (DCD) is a chronic neurodevelopmental disorder that results in difficulty in motor coordination observed in school-going children that interferes with classroom performance. Suspected DCD (S-DCD) children may show poor motor, as well as academic performance at school, and hence the present study aimed to find out the prevalence of S-DCD in children of age 5-10 years in central India and to find its association with preterm and/or low birth weight (LBW). Method: A total of 716 normal school-going children of age 5-10 years (both genders) were included in the study from four schools of the city by stratified sampling method. Children with any diagnosed neurological, orthopedic, rheumatologic, metabolic, cardiopulmonary, or psychological disorders were excluded. Data was collected using the parent-administered Developmental Coordination Disorder Questionnaire-2007 (DCDQ'07) and a parent/caregiver proforma. Children were sorted into three age subgroups (5-7.11 years, 8-9.11 years and 9-9.11 years). Result: Prevalence of S-DCD in 5-7.11 years (21.5%), 8-9.11years (23.9%) and is highest in 10-10.11 years (30.6%). Preterm children showed a higher prevalence of S-DCD (preterm: 29.54%, term: 23.10%). Children with LBW also showed a higher prevalence of S-DCD (30.15%) and among normal birth weight (21.43%). In children with both preterm and LBW history, the prevalence of suspected DCD was found to be 51.72%. Conclusion: Prevalence of suspected DCD was found to be 23.9% in the 5-10 years age group. It was also observed that S-DCD is strongly associated with preterm, as well as low birth weight in children of age 5-10 years.

Utility of Bedside Ultrasound Measurement of Optic Nerve Sheath Diameter as a Screening Tool for raised Intracranial Pressure in Neurocritical Care Prospective Observational Study.

Intracranial pressure (ICP) needs to be monitored in neurocritical patients. There is a need for portable bedside optic nerve ultrasound (ONUS) for early diagnosis to initiate the measures to reduce ICP Objective: To find the utility of bedside ONUS to diagnose raised ICP in neurocritical care. Materials and methods: After approval from the ethical committee, a prospective observational study was conducted. Optic nerve sheath diameter (ONSD) was measured in two groups: control group patients with neurological symptoms but computed tomography (CT)/magnetic resonance imaging (MRI) not suggestive of raised ICP, and second was study group patients with neurological symptoms and CT/MRI suggestive of elevated ICP Result: In patients with normal ICP, the mean ONSD in females was 4.47mm, and in males was 4.66mm. In patients with raised ICP, the mean ONSD in females was 6.45 ± 0.78 mm, and in males was 6.33 ± 0.70 mm. Regarding the correlation between Glasgow coma scale (GCS) and mean ONSD parameters, the coefficient of correlation (R) is 0.14; thus, there is a weak negative correlation. In our study, no difference was observed in raised mean ONSD in patients with different diagnoses. At a cut-off value of >4.8 mm, the sensitivity and specificity are 100% to diagnose raised ICP. Conclusion: Optic nerve sheath diameter (ONSD) is a reliable, rapid bedside screening tool in the Emergency Department/Critical Care/Operation Theatre to diagnose raised ICP. In order to keep a record of trends in ICP, we need to measure ONSD frequently. There was no correlation between GCS and ONSD measurement.

Ulinastatin Add-on to Standard of Care in Critically Ill COVID-19 Patients: A Multicenter, Retrospective Study.

AIM: To assess the impact on 30-day mortality with ulinastatin (ULI) used as add-on to standard of care (SOC) compared to SOC alone in coronavirus disease (COVID-19) patients requiring admission to the intensive care unit (ICU). MATERIALS AND METHODS: In this multicentric, retrospective study, we collected data on clinical, laboratory, and outcome parameters in patients with COVID-19. Thirty-day mortality outcome was compared among patients treated with SOC alone and ULI used as add-on to SOC. Odds ratio (OR) and 95% confidence intervals (CI) were determined to identify the predictors of 30-day mortality. RESULTS: Ninety-four patients were identified and enrolled in both groups with comparable baseline parameters. On univariate analysis, 30-day mortality was significantly lower in ULI plus SOC group than SOC alone group (36.2 vs 51.1%, OR 0.54, 95% CI 0.30-0.97, p = 0.040). The effect on mortality was more pronounced in patients who did not require intubation (10.9 vs 34.0%, OR 0.24, 95% CI 0.09-0.66, p = 0.006) and with early administration (within 72 hours of admission) of ULI (30.7 vs 57.9%, OR 0.32, 95% CI 0.11-0.91, p = 0.032). On multivariate analysis, only intubation predicted mortality (adjusted OR 10.13, 95% CI 3.77-27.25, p<0.0001) and the effect of ULI on survival was not significant (adjusted OR 0.58, 95% CI 0.22-1.52, p = 0.270). CONCLUSION: Given the limited options for COVID-19 patients treated in ICU, early administration of ULI may be helpful, especially in patients not requiring intubation to improve the outcomes. Further, a large, randomized study is warranted to confirm these findings.

EMTome: a resource for pan-cancer analysis of epithelial-mesenchymal transition genes and signatures.

BACKGROUND: The epithelial-mesenchymal transition (EMT) enables dissociation of tumour cells from the primary tumour mass, invasion through the extracellular matrix, intravasation into blood vessels and colonisation of distant organs. Cells that revert to the epithelial state via the mesenchymal-epithelial transition cause metastases, the primary cause of death in cancer patients. EMT also empowers cancer cells with stem-cell properties and induces resistance to chemotherapeutic drugs. Understanding the driving factors of EMT is critical for the development of effective therapeutic interventions. METHODS: This manuscript describes the generation of a database containing EMT gene signatures derived from cell lines, patient-derived xenografts and patient studies across cancer types and multiomics data and the creation of a web-based portal to provide a comprehensive analysis resource. RESULTS: EMTome incorporates (i) EMT gene signatures; (ii) EMT-related genes with multiomics features across different cancer types; (iii) interactomes of EMT-related genes (miRNAs, transcription factors, and proteins); (iv) immune profiles identified from The Cancer Genome Atlas (TCGA) cohorts by exploring transcriptomics, epigenomics, and proteomics, and drug sensitivity and (iv) clinical outcomes of cancer cohorts linked to EMT gene signatures. CONCLUSION: The web-based EMTome portal is a resource for primary and metastatic tumour research publicly available at www.emtome.org .

Increase in Cadaver Organ Donation Rate at a Tertiary Care Hospital: 23 Years of Experience.

BACKGROUND: Transplantation of Human Organ Act was passed in India in 1994 to streamline organ donation and transplantation activities. It is time to retrospect ourselves and analyze the method to increase organ donation. TYPE OF STUDY: Retrospective observational analysis. OBJECTIVES: To evaluate the change in organ donation rate and reasons for changes in rates. SUBJECTS: Brainstem dead declared patients whose family consented for organ donations in the last 23 years (1997-2019) at Ruby Hall Clinic, Pune, India. MATERIALS AND METHODS: Retrospectively demographic data of the brainstem dead declared donors, the primary diagnoses, comorbidities, and the complete data of their management till organ retrieval was assessed. RESULTS: One hundred cases in the age group 15-75 years (mean 41.6 ± 15.3 years) of brainstem death consented for organ donation were retrospectively studied. The period was divided into two groups, group I and group II included study duration from 1997 to 2013 and from 2013 to 2019 respectively. During the entire period, though the major cause of donor death remained road traffic accidents (RTA) in both the groups (84.21% till 2013 vs 48.15% after 2013), the proportion of donors declared brain dead due to RTA dipped significantly after 2013 (p = 0.004) and the non-RTA causes of brain dead contributed more than RTA causes (51.85% non-RTA vs 48.15% RTA). The major contributor among non-RTA causes was intracranial bleeds (5.26% before 2013 vs 33.33% after 2013, p = 0.014). Compared to the previous 17 years (from 1997) there were more than fourfold rise in the rate of transplantation in the last 6 years (2014-2019) at our institute. Kidneys were retrieved from 90% donors followed by cornea 84%, liver 65%, heart 22%, skin 7%, lungs 6%, and pancreas 5%. CONCLUSION: We have observed that the cadaveric organ donation rate significantly improved after 2013. Reasons might be widening of the donor pool by the selection of more of non-RTA brain death donors over RTA, acceptability of elderly population donor (>60 years) by our transplant teams, early identification of potential organ donor, and better protocol-based management of the cadaver organ donor. HOW TO CITE THIS ARTICLE: Zirpe KG, Suryawanshi P, Gurav S, Deshmukh A, Pote P, Tungenwar A, et al. Increase in Cadaver Organ Donation Rate at a Tertiary Care Hospital: 23 Years of Experience. Indian J Crit Care Med 2020;24(9):804-808.

Incidence of Medication Error in Critical Care Unit of a Tertiary Care Hospital: Where Do We Stand?

BACKGROUND: Medication error in developed countries is of primary concern when there is a question of adversity to a patient's health, but in developing countries like India, it is just a term and its significance is undervalued. The incidence of medication error is essential to estimate the proper medical care provided in the healthcare system. OBJECTIVE: The main objective of the study is to determine the incidences of medication error in critical care unit and to evaluate its risk outcomes. MATERIALS AND METHODS: This is a prospective observational study conducted over a period of 6 months in a critical care unit of a tertiary care hospital. Medication chart review method was opted for data collection. The medication errors were mainly classified as prescription, transcription, indenting, dispensing, and administration error. A total of 6,705 charts were reviewed. The NCCMERP risk index was used to evaluate the outcome of errors. RESULTS: Of the total 6,705 charts, 410 medication errors were found, i.e., 6.11%. The most common error is transcription error that constitutes 44.1% of the total errors, followed by prescription error 40%, and administration error 14%. The frequency of indenting and dispensing errors is negligible with 1.5% and 0.5%, respectively. The main causes of medication errors are due to incomplete prescription 50.2% and wrong doses 22.9%. In drug class, antibiotics and antihypertensive agents are most prone to medication error. About 87.1% errors belonged to the Category B of National Coordinating Council for Medication Error Reporting and Prevention risk index. CONCLUSION: Majority of the errors are transcription errors followed by prescription and administration errors. Consultant doctors have to be more vigilant during prescribing and verifying the medication charts. Clinical pharmacists should act as a checkpoint at each step of medication process to identify and prevent medication errors. HOW TO CITE THIS ARTICLE: Zirpe KG, Seta B, Gholap S, Aurangabadi K, Gurav SK, Deshmukh AM, et al. Incidence of Medication Error in Critical Care Unit of a Tertiary Care Hospital: Where Do We Stand? Indian J Crit Care Med 2020;24(9):799-803.

A possible role for epigenetic feedback regulation in the dynamics of the epithelial-mesenchymal transition (EMT).

The epithelial-mesenchymal transition (EMT) often plays a critical role in cancer metastasis and chemoresistance, and decoding its dynamics is crucial to design effective therapeutics. EMT is regulated at multiple levels-transcriptional, translational, protein stability and epigenetics; the mechanisms by which epigenetic regulation can alter the dynamics of EMT remain elusive. Here, to identify the possible effects of epigenetic regulation in EMT, we incorporate a feedback term in our previously proposed model of EMT regulation of the miR-200/ZEB/miR-34/SNAIL circuit. This epigenetic feedback that stabilizes long-term transcriptional activity can alter the relative stability and distribution of states in a given cell population, particularly when incorporated in the inhibitory effect on miR-200 from ZEB. This feedback can stabilize the mesenchymal state, thus making transitions out of that state difficult. Conversely, epigenetic regulation of the self-activation of ZEB has only minor effects. Our model predicts that this effect could be seen in experiments, when epithelial cells are treated with an external EMT-inducing signal for a sufficiently long period of time and then allowed to recover. Our preliminary experimental data indicates that a chronic TGF-ß exposure gives rise to irreveversible EMT state; i.e. unable to reverse back to the epithelial state. Thus, this integrated theoretical-experimental approach yields insights into how an epigenetic feedback may alter the dynamics of EMT.

Impact of "Stroke Code"-Rapid Response Team: An Attempt to Improve Intravenous Thrombolysis Rate and to Shorten Door-to-Needle Time in Acute Ischemic Stroke.

OBJECTIVE: "Stroke code" (SC) implementation in hospitals can improve the rate of thrombolysis and the timeline in care of stroke patient. MATERIALS AND METHODS: A prospective data of patients treated for acute ischemic stroke (AIS) after implementation of "SC" (post-SC era) were analyzed (2015-2016) and compared with the retrospective data of patients treated in the "pre-SC era." Parameters such as symptom-to-door, door-to-physician, door-to-imaging, door-to-needle (DTN), and symptom-to-needle time were calculated. The severity of stroke was calculated using the National Institutes of Health Stroke Score (NIHSS) before and after treatment. RESULTS: Patients presented with stroke symptoms in pre- and post-SC era (695 vs. 610) and, out of these, patients who came in window period constituted 148 (21%) and 210 (34%), respectively. Patients thrombolyzed in pre- and post-SC era were 44 (29.7%) and 65 (44.52%), respectively. Average DTN time was 104.95 min in pre-SC era and reduced to 67.28 min (P < 0.001) post-SC implementation. Percentage of patients thrombolyzed within DTN time ≤60 min in pre-SC era and SC era was 15.90% and 55.38%, respectively. CONCLUSION: Implementation of SC helped us to increase thrombolysis rate in AIS and decrease DTN time.

Cancer stem cell metabolism: a potential target for cancer therapy.

Cancer Stem cells (CSCs) are a unipotent cell population present within the tumour cell mass. CSCs are known to be highly chemo-resistant, and in recent years, they have gained intense interest as key tumour initiating cells that may also play an integral role in tumour recurrence following chemotherapy. Cancer cells have the ability to alter their metabolism in order to fulfil bio-energetic and biosynthetic requirements. They are largely dependent on aerobic glycolysis for their energy production and also are associated with increased fatty acid synthesis and increased rates of glutamine utilisation. Emerging evidence has shown that therapeutic resistance to cancer treatment may arise due to dysregulation in glucose metabolism, fatty acid synthesis, and glutaminolysis. To propagate their lethal effects and maintain survival, tumour cells alter their metabolic requirements to ensure optimal nutrient use for their survival, evasion from host immune attack, and proliferation. It is now evident that cancer cells metabolise glutamine to grow rapidly because it provides the metabolic stimulus for required energy and precursors for synthesis of proteins, lipids, and nucleic acids. It can also regulate the activities of some of the signalling pathways that control the proliferation of cancer cells.This review describes the key metabolic pathways required by CSCs to maintain a survival advantage and highlights how a combined approach of targeting cellular metabolism in conjunction with the use of chemotherapeutic drugs may provide a promising strategy to overcome therapeutic resistance and therefore aid in cancer therapy.

Problems and limitations in thrombolysis of acute stroke patients at a tertiary care center.

AIM: (1) To evaluate the number of patients thrombolysed within 1 h of arrival to emergency room (ER) (2) To identify reasons for delay in thrombolysis of acute stroke patients. MATERIALS AND METHODS: All patients admitted to ER with symptoms suggestive of stroke from January 2011 to November 2013 were studied. Retrospective data were collected to evaluate ER to needle (door to needle time [DTNt]) time and reasons for delay in thrombolysis. The parameters studied (1) onset of symptoms to ER time, (2) ER to imaging time (door to imaging time [DTIt]), (4) ER to needle time (door to needle) and (5) contraindications for thrombolysis. RESULTS: A total of 695 patients with suspected stroke were admitted during study period. 547 (78%) patients were out of window period. 148 patients (21%, M = 104, F = 44) arrived within window period (<4.5 h.). 104 (70.27%) were contraindicated for thrombolysis. Majority were intracerebral bleeds. 44 (29.7%) were eligible for thrombolysis. 7 (15.9%) were thrombolysed within 1 h. The mean time for arrival of patients from onset of symptoms to hospital (symptom to door) 83 min (median - 47). The mean door to neuro-physician time (DTPt) was 32 min (median - 15 min). The mean DTIt was 58 min (median - 50 min). The mean DTNt 104 (median - 100 min). CONCLUSION: Reasons for delay in thrombolysis are: Absence of stroke education program for common people. Lack of priority for triage and imaging for stroke patients.

Normal values of functional reach and lateral reach tests in children with knee hypermobility.

PURPOSE: To measure values for functional reach (FR) and lateral reach (LR) in school children with knee joint hypermobility (KJH), and to examine the correlation of anthropometric measures and KJH with FR and LR values. METHODS: A total of 140 children aged 6 to 12 years with typical development (TD) and bilateral KJH of greater than 10° hyperextension participated. Three successive trials of FR and LR tests in standing position with feet shoulder width apart were performed, and the mean of the 3 trials was calculated. Analysis of variance and Bonferroni tests were used to analyze correlation and association of FR and LR values with KJH angle and anthropometric data, respectively, with a 95% confidence interval. RESULTS: In school-aged children with TD, height and KJH contributed significantly to the FR values in both genders, whereas height contributed for LR values among girls alone. CONCLUSIONS: Height and KJH affect children's scores on the FR and LR tests of balance.

Enrichment of Cancer Stem Cells in a Tumorsphere Assay.

Cancer stem cells (CSCs) are a small subpopulation of self-renewing cancer cells that are present within tumors. CSCs possess tumor initiation potential as well as the ability to resist toxic compounds and chemotherapeutic agents through the upregulation of drug efflux transporters, DNA repair pathways, and survival cascades. Accumulating evidence suggests that CSCs are responsible for tumor relapse and resistance to chemotherapeutic agents and that targeting CSCs is critical to inhibition of cancer progression. Therefore, isolation and characterization of CSCs is important in studying tumor initiation and progression. In this chapter, we provide a detailed method for the identification and isolation of CSCs.

Limiting Dilution Tumor Initiation Assay: An In Vivo Approach for the Study of Cancer Stem Cells.

Cancer stem cells (CSCs) are a small subpopulation of self-renewing cancer cells that are present within tumors. Calculating the frequency of tumor-initiating cells is important in the assessment of the number of CSCs present in a cell population. In this chapter, we present a protocol developed for quantification of CSCs from breast cancer tumors that can be adapted to CSCs from other types of tumors.

In Vitro Quantification of Cancer Stem Cells Using a Mammosphere Formation Assay.

Cancer stem cells (CSCs) are a small subpopulation of self-renewing cancer cells that are present within tumors. In this chapter, we provide a detailed method for the quantification of CSCs in vitro through mammosphere formation.

Normal values of functional reach and lateral reach tests in Indian school children.

PURPOSE: To establish normal values for functional reach (FR) and lateral reach (LR) in school children and to study the correlation of anthropometric measures with FR and LR values and the association between FR and LR scores. METHODS: A total of 350 children aged 6 to 12 years were randomly selected. Three successive trials of FR and LR with the child standing with feet shoulder width apart were performed and the mean of 3 trials was calculated. RESULTS: Normal values of FR and LR were obtained. Pearson product moment correlation was used to examine the association of FR and LR to age, gender, and anthropometric measures. Stepwise regression analysis was done to obtain normal values of FR and LR with respect to height. Functional reach values significantly predicted LR values and vice versa. CONCLUSION: Normal mean values of FR and LR range from 22.7 cm to 37 cm and 16.3 cm to 22.5 cm, respectively. Height significantly correlates with both FR and LR.

Breast cancer dormancy: need for clinically relevant models to address current gaps in knowledge.

Breast cancer is the most commonly diagnosed cancer in the USA. Although advances in treatment over the past several decades have significantly improved the outlook for this disease, most women who are diagnosed with estrogen receptor positive disease remain at risk of metastatic relapse for the remainder of their life. The cellular source of late relapse in these patients is thought to be disseminated tumor cells that reactivate after a long period of dormancy. The biology of these dormant cells and their natural history over a patient's lifetime is largely unclear. We posit that research on tumor dormancy has been significantly limited by the lack of clinically relevant models. This review will discuss existing dormancy models, gaps in biological understanding, and propose criteria for future models to enhance their clinical relevance.

Functional dissection of the catalytic carboxyl-terminal domain of origin recognition complex subunit 1 (PfORC1) of the human malaria parasite Plasmodium falciparum.

Origin recognition complex subunit 1 (ORC1) is essential for DNA replication in eukaryotes. The deadly human malaria parasite Plasmodium falciparum contains an ORC1/CDC6 homolog with several interesting domains at the catalytic carboxyl-terminal region that include a putative nucleoside triphosphate-binding and hydrolysis domain, a putative PCNA-interacting-protein (PIP) motif, and an extreme C-terminal region that shows poor homology with other ORC1 homologs. Due to the unavailability of a dependable inducible gene expression system, it is difficult to study the structure and function of essential genes in Plasmodium. Using a genetic yeast complementation system and biochemical experiments, here we show that the putative PIP domain in ORC1 that facilitates in vitro physical interaction with PCNA is functional in both yeast (Saccharomyces cerevisiae) and Plasmodium in vivo, confirming its essential biological role in eukaryotes. Furthermore, despite having less sequence homology, the extreme C-terminal region can be swapped between S. cerevisiae and P. falciparum and it binds to DNA directly, suggesting a conserved role of this region in DNA replication. These results not only provide us a useful system to study the function of the essential genes in Plasmodium, they help us to identify the previously undiscovered unique features of replication proteins in general.

Epigenetic demethylation of sFRPs, with emphasis on sFRP4 activation, leading to Wnt signalling suppression and histone modifications in breast, prostate, and ovary cancer stem cells.

The expression and levels of secreted frizzled-related proteins (sFRPs), important Wnt signalling antagonists, have been reported to be reduced in various cancers, and are associated with disease progression and poor prognosis. During tumour development, all sFRP (1, 2, 3, 4, and 5) genes are hypermethylated, causing transcriptional silencing. sFRPs have an ability to sensitize tumour cells to chemotherapeutic drugs, enhancing cell death. Reduced Wnt signalling is associated with loss of cancer stem cell (CSC) viability. We investigated the possible involvement of methylation-mediated silencing of the sFRP gene family in CSCs derived from breast, prostate, and ovarian tumour cell lines. Real-time RT-PCR studies indicated that loss or downregulation of sFRP (1-5) expression in tumours is associated with promoter hypermethylation. Additionally, CSCs derived from all tumour cell lines with sFRP (1-5) promotor hypermethylation expressed sFRP (1-5) mRNA after treatment with 5-Azacytidine (5-Aza), especially sFRP4, implying that DNA methylation is the predominant epigenetic mechanism for sFRP (1-5) silencing. Furthermore, post-translational modification (PTM) in total and histone proteins was observed post 5-Aza and sFRP4 treatment. Protein levels of Wnt downstream signalling components (GSK3ß, active ß-catenin, and phospho ß-catenin) and epigenetic factors of histones (acetyl histone H3, and H3K27me3) affecting PTM were analysed. Our findings suggest that downregulation of sFRP4 expression in endocrine-related cancers can be attributed to aberrant promoter hypermethylation in conjugation with histone modification, and indicate the important role of methylation-induced gene silencing of sFRP4 in survival and proliferation of CSCs derived from these cancers.