Ataxia-Telangiectasia Mutated (ATM) gene signaling pathways in human cancers and their therapeutic implications.

Cancer is a multifaceted disease driven by abnormal cell growth and poses a significant global health threat. The multifactorial causes, differences in individual susceptibility to therapeutic drugs, and induced drug resistance pose major challenges in addressing cancers effectively. One of the most important aspects in making cancers highly heterogeneous in their physiology lies in the genes involved and the changes occurring to some of these genes in malignant conditions. The Genetic factors have been implicated in the oncogenesis, progression, responses to treatment, and metastasis. One such gene that plays a key role in human cancers is the mutated form of the Ataxia-telangiectasia gene (ATM). ATM gene located on chromosome 11q23, plays a vital role in maintaining genomic stability. Understanding the genetic basis of A-T is crucial for diagnosis, management, and treatment. Breast cancer, lung cancer, prostate cancer, and gastric cancer exhibit varying relationships with the ATM gene and influence their pathways. Targeting the ATM pathway proves promising for enhancing treatment effectiveness, especially in conjunction with DNA damage response pathways. Analyzing the therapeutic consequences of ATM mutations, especially in these cancer types facilitates the approaches for early detection, intervention, development of personalized treatment approaches, and improved patient outcomes. This review emphasizes the role of the ATM gene in various cancers, highlighting its impact on DNA repair pathways and therapeutic responses.

Assessment of genome mutation analysis for tumor-informed detection of circulating tumor DNA in patients with breast cancer.

INTRODUCTION: Breast cancer, one of the most aggressive types of cancer, poses significant challenges for diagnosis and treatment. Emerging as a promising biomarker, circulating tumor DNA (ctDNA) can be used to identify and monitor disease risk. This study sought to examine the impact of mutations in various genes on the progression of breast cancer. Genetic variants associated with breast cancer have been examined in individuals diagnosed with the disease worldwide. METHODS: Fifty female participants underwent breast cancer testing. Sanger sequencing was used to analyze peripheral blood DNA from these individuals to detect disease-causing mutations in the BRCA1, BRCA2, PTEN, TP53, and ATM genes. Genetic alterations linked to breast cancer were screened and the findings were compared with those of tumor genes. RESULTS: The development of hereditary/early onset breast cancer in this study was significantly associated with mutations in ATM, PTEN, TP53, and BRCA1/BRCA2, according to the analysis of sequencing data. CONCLUSION: This study demonstrates the feasibility of analyzing ctDNA in patients with breast cancer (BC) undergoing palliative treatment using an SS-based technique.

Microfluidics as diagnostic tools.

The challenges in the management of human diseases are largely determined by the precision, speed and ease of diagnostic procedures available. Developments in biomedical engineering technologies have greatly helped in transforming human health care, especially for disease diagnosis which in turn lead to better patient outcomes. One such development is in the form of microfluidic chip technology which has transformed various aspects of human health care. We present in this review, a comprehensive account on the utility of microfluidic chip technologies for the diagnosis of autoimmune disorders, cardiovascular diseases (CVDs), infectious diseases, and neurodegenerative conditions. We have included the diseases posing global threat such as rheumatoid arthritis, diabetes, pernicious anemia, tuberculosis, COVID-19, influenza, alzheimer's, multiple sclerosis, and epilepsy. Apart from discussing the ways of microfluidic chip in diagnosis, we included a section presenting electrochemical, electrical, optical, and acoustic detection technologies for the precise diagnosis of CVDs. Microfluidics platforms have thus revolutionized novel capabilities in addressing the requirements of point-of-care diagnostics enabling miniaturization by integrating multiple laboratory functions into a single chip resulting in "one flow - one solution" systems. Hence, the precision and early diagnoses of diseases are now possible due to the advancements of microfluidics-based technology.

Implications and progression of peroxiredoxin 2 (PRDX2) in various human diseases.

Peroxiredoxin 2 (PRDX2), a characteristic 2-Cys enzyme is one of the foremost effective scavenger proteins against reactive oxygen species (ROS) and hydrogen peroxide (H2O2) defending cells against oxidative stress. Dysregulation of this antioxidant raises the quantity of ROS and oxidative stress implicated in several diseases. PRDX2 lowers the generation of ROS that takes part in controlling several signalling pathways occurring in neurons, protecting them from stress caused by oxidation and an inflammatory harm. Depending on the aetiological variables, the kind of cancer, and the stage of tumour development, PRDX2 may behave either as an onco-suppressor or a promoter. However, overexpression of PRDX2 may be linked to the development of numerous cancers, including those of the colon, cervix, breast, and prostate. PRDX2 also plays a beneficial effect in inflammatory diseases. PRDX2 being a thiol-specific peroxidase, is known to control proinflammatory reactions. The spilling of PRDX2, on the other hand, accelerates cognitive impairment following a stroke by triggering an inflammatory reflex. PRDX2 expression patterns in vascular cells tend to be crucial to its involvement in cardiovascular diseases. In vascular smooth muscle cells, if the protein tyrosine phosphatase is restricted, PRDX2 could avoid the neointimal thickening which relies on platelet derived growth factor (PDGF), a vital component of vascular remodelling. A proper PRDX2 balance is therefore crucial. The imbalance causes a number of illnesses, including cancers, inflammatory diseases, cardiovascular ailments, and neurological and neurodegenerative problems which are discussed in this review.

Prospectives and retrospectives of microfluidics devices and lab-on-A-chip emphasis on cancer.

Microfluidics is a science and technology that deals with the concept of "less sample-to-more precision" enabling portable device development via fabrication for in vitro analysis. On evolution, microfluidic system lead to the development of Organ-on-chip where recapitulation of organ's functionality and pathophysiological response can be performed under controlled environment. Further microfluidic-based "Lab-on-chip" device, a versatile innovation credited for its number of parameters that has capability to leverage next-generation companion of medicines. This emulsion science has enormous practise in the field of regenerative medicine, drug screening, medical diagnosis and therapy for accuracy in results. In this era of personalized medicine, getting precise tools for applying these theranostics is crucial. Oncological theranostics create a new gateway to develop precision in personalized medicine for cancer, where microfluidic chips are involved in diagnosis and therapy of various cancers using biomarkers for thyroid, lung cancers, and assay based for breast, circulating tumor cells and colorectal cancers and nanoparticles for ovarian cancer. This review shows more comprehensive approach to the state of art with respect to microfluidic devices in cancer theranostics.

Biomarkers and biosensors for early cancer diagnosis, monitoring and prognosis.

Cancers continue to be of major concern due to their serious global socioeconomic impact, apart from the continued increase in the incidence of various cancer types. A major challenge that this disease poses is due to the low "early detection" rates which limit the therapeutic outcomes for the affected individuals. Current research has highlighted the discovering biomarkers that help in early cancer detection and the development of technologies for the detection and quantification of such biomarkers. Biomarkers range from proteins to nucleic acids, and can be specific to a particular cancer type. Detection and quantification of such biomarkers at low levels from biological samples is being made possible by the advent of developing biosensors and by using biomedical engineering technologies such as tumor-on-a-chip models. Here, we present biomarkers that can be helpful for the early detection of breast, colorectal, esophageal, lung, liver, ovarian, and prostate cancer. In addition, we discuss the potential of circulating tumor cell DNA (ctDNA) as an early diagnostic marker. Finally, biosensors available for the detection of cancer biomarkers, which is a recent advancement in this area of research, are discussed.

Significance of cell culture phases for utilizing 3D cultures of A549 cells for in vitro studies.

Three-dimensional (3D) culture systems of human cancer cell lines have become popular experimental models for a variety of applications including drug screening. It is understood that the 2D and 3D cultures of the same cell line behave differently in several aspects. One such difference is in the duration of cell culture phases (the lag, log, plateau and the decline stages). We obtained 3D cultures of A549 cells on agarose hydrogels. We observed and compared the morphological differences in the progression of 2D and 3D cultures of A549 cells in a time-dependent manner. The morphological features along with the cell counts and viabilities obtained for the 2D and 3D cultures at different time intervals clearly indicate that the cell culture phases occurred as more extended one for the 3D cultures compared to that of the 2D counterparts. The plateau stage for the 2D and 3D cultures occurred at 48 and 69 h, respectively. Such cell culture phase durations can be different for different cell lines as a function of their doubling times. We propose that the cell culture phase durations for any cell line should be first established before using them for drug testing or for studies involving toxicity to obtain useful results from 3D cell cultures. Also, we propose that the late-exponential (lag) phase of 3D cultures of cancer cell lines is the most ideal one for drug testing owing to the various optimal features of the aggregates in this cell culture phase.

Role of homeobox d10 gene targeted signaling pathways in cancers.

Homeobox D10 (HOXD10) is a transcription factor from the homeobox gene family that controls cell differentiation and morphogenesis throughout development.Due to their functional interaction, changes in HOXD10 gene expression might induce tumors. This narrative review focuses on how and why the dysregulation in the signaling pathways linked with HOXD10 contributes to the metastatic development of cancer. Organ development and tissue homeostasis need highly conserved homeotic transcription factors from homeobox (HOX) genes. Their dysregulation disrupts regulatory molecule action, causing tumors. The HOXD10 gene is upregulated in breast, gastric, hepatocellular, colorectal, bladder, cholangiocellular carcinoma and prostate cancer. Tumor signaling pathways are affected by HOXD10 gene expression changes. This study examines HOXD10-associated signaling pathway dysregulation, which may alter metastatic cancer signaling. In addition, the theoretical foundations that alter HOXD10-mediated therapeutic resistance in malignancies has been presented. New cancer therapy methods will be simpler to develop with the newly discovered knowledge. This review showed that HOXD10 may be a tumor suppressor gene and a new cancer treatment target signaling pathway.

Prospectives of mirna gene signaling pathway in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is one of the destructive breast cancer subtypes which cannot be treated by current therapies, which is characterized by the lack of estrogen (ER), Progesterone (PR), and Human epidermal receptor (HER2). The treatment for this chemotherapy or radiotherapy and surgery are such treatments and also novel biomarkers or treatment targets can quickly require to improve the outcome of the disease. MicroRNAs are the most popular and offer prospects for TNBC diagnosis and therapy. Some of the miRNAs implicated in THBCs are miR-17-5p, miR-221-3p, miR-26a, miR-136-5p, miR-1296, miR-145, miR-4306, miR-508-5p, miR-448, miR-539, miR-211-5p and miR-218. Potential MiRNAs and their signaling pathways that can be utilized for the diagnosis of TNBC are miR-155, miR-182-5p, miR-9-1-5p, miR-200b, miR-200a, miR-429, miR-195, miR-145-5p, miR-506, and miR-22-3p. miRNAs with known functions as tumor suppressors include miR-1-3p, miR-133a-3p, miR-655, miR-206, miR-136, miR-770, miR-148a, miR-197-3p, miR-137, and miR-127-3p. Analysis of genetic biomarkers, such as miRNAs in TNBC, upholds the pertinence in the diagnosis of the disease. The aim of the review was to clarify the different types of miRNAs characters in TNBC. Recent reports suggest an important role of miRNAs in tumor metastasis. We review here the important miRNAs and their signaling pathways implicated in the oncogenesis, progression, and metastasis of TNBCs.

Recent and advanced therapy for oral cancer.

Oral cancer is a common and deadly kind of tissue invasion, has a high death rate, and may induce metastasis that mostly affects adults over the age of 40. Most in vitro traditional methods for studying cancer have included the use of monolayer cell cultures and several animal models. There is a worldwide effort underway to reduce the excessive use of laboratory animals since, although being physiologically adequate, animal models rarely succeed in exactly mimicking human models. 3D culture models have gained great attention in the area of biomedicine because of their capacity to replicate parent tissue. There are many benefits to using a drug delivery approach based on nanoparticles in cancer treatment. Because of this, in vitro test methodologies are crucial for evaluating the efficacy of prospective novel nanoparticle drug delivery systems. This review discusses current advances in the utility of 3D cell culture models including multicellular spheroids, patient-derived explant cultures, organoids, xenografts, 3D bioprinting, and organoid-on-a-chip models. Aspects of nanoparticle-based drug discovery that have utilized 2D and 3D cultures for a better understanding of genes implicated in oral cancers are also included in this review.

Gene expressions and their significance in organoid cultures obtained from breast cancer patient-derived biopsies.

Gene expression changes are one of the hallmarks of malignant cells and such changes in specific genes have been identified for a variety of human cancers. Such an association in gene expression changes becomes very significant for breast cancers due to the genetic heterogeneity seen in such cancers. It is due to such genetic implication that breast cancers are classified into several subtypes; based on the expression and the magnitude of expression of estrogen and progesterone receptor genes. Changes in the expression of ERBB2, ESR1, PLAU, MUC1, PGR, and TP53 are implicated in breast cancers. Of the various models available for cancer research, organoid cultures from patient-derived biopsies are being considered as the most relevant for invitro testing. Organoid cultures derived from patient biopsies mitigate several limitations of other commonly available models such as cancer cell lines. Such organoids retain the functional physiology of solid tumors which include gene expression. Also, utilizing patient derived organoids for in vitro testing paves way for personalized medicine which greatly enhances the effectiveness of cancer therapy for individuals. We present the genes implicated in breast cancers, the ways in which organoids can be derived from breast cancer biopsies and their applications for gene expression studies.

Development of patient derived organoids for cancer drug screening applications.

Cancer is a disease characterised by abnormal cell growth that can invade or spread to other regions of the body. Organoids are three-dimensional ex vivo tissue cultures made from embryonic stem cells, induced pluripotent stem cells, progenitor cells or tissue that serve as a physiological model for cancer research. These are designed to recapitulate the in vivo properties of tumours. Importantly, effective recapitulation of the structure of tissues and function is believed to predict patient response, allowing for the creation of personalised therapy in a timely manner that may be used in the clinic. This Review discusses the pre-clinical model and different types of human organoids as models for the development of high throughput drug screening and also aims to highlight how organoids are shaping the future of cancer research.

Approaches for Head and Neck Cancer Research - Current Status and the Way Forward.

Head and neck cancers (HNCs) are seeing an increasing trend in their prevalence among both genders and are the seventh most common cancer type occurring at the global level. Studies addressing both the cancer cell physiology and individual differences in response to a specific treatment modality should be understood for arriving at effective treatment and management of the HNCs. In this article, we discuss the trends in HNC research and their various approaches starting from 2D in vitro models, which are the traditional experimental materials to recently established Cancer-Tissue Originated Spheroids (CTOS) distinctly contributing towards personalized or precision medicine.

Devices and techniques used to obtain and analyze three-dimensional cell cultures.

Cell cultures are indispensable for both basic and applied research. Advancements in cell culture and analysis increase their utility for basic research and translational applications. A marked development in this direction is advent of three-dimensional (3D) cultures. The extent of advancement in 3D cell culture methods over the past decade has warranted referring to a single cell type being cultured as an aggregate or spheroid using simple scaffolds as "traditional." In recent years, the development of "next-generation" devices has enabled cultured cells to mimic their natural environments much better than the traditional 3D culture systems. Automated platforms like chip-based devices, magnetic- and acoustics-based assembly devices, di-electrophoresis (DEP), micro pocket cultures (MPoC), and 3D bio-printing provide a dynamic environment compared to the rather static conditions of the traditional simple scaffold-based 3D cultures. Chip-based technologies, which are centered on principles of microfluidics, are revolutionizing the ways in which cell culture and analysis can be compacted into table-top instruments. A parallel evolution in analytical devices enabled efficient assessment of various complex physiological and pathological endpoints. This is augmented by concurrent development of software enabling rapid large-scale automated data acquisition and analysis like image cytometry, elastography, optical coherence tomography, surface-enhanced Raman scattering (SERS), and biosensors. The techniques and devices utilized for the purpose of 3D cell culture and subsequent analysis depend primarily on the requirement of the study. We present here an in-depth account of the devices for obtaining and analyzing 3D cell cultures.

Factors, mechanisms and implications of chromatin condensation and chromosomal structural maintenance through the cell cycle.

A series of well-orchestrated events help in the chromatin condensation and the formation of chromosomes. Apart from the formation of chromosomes, maintenance of their structure is important, especially for the cell division. The structural maintenance of chromosome (SMC) proteins, the non-SMC proteins and the SMC complexes are critical for the maintenance of chromosome structure. While condensins have roles for the DNA compaction, organization, and segregation, the cohesin functions in a cyclic manner through the cell cycle, as a "cohesin cycle." Specific mechanisms maintain the architecture of the centromere, the kinetochore and the telomeres which are in tandem with the cell cycle checkpoints. The presence of chromosomal territories and compactness differences through the length of the chromosomes might have implications on selective susceptibility of specific chromosomes for induced genotoxicity.

The culture conditions and outputs from breast cancer cell line in vitro experiments.

One of the major cancer types that have gained significant importance globally is the breast cancer due to its socio-economic impact. Breast cancer research is an area of considerable importance and several types of material are available for research applications. These include cancer cell lines which can be utilized in several ways. Cell lines are convenient to use and recently about 84 human breast cancer cell lines were classified by molecular sub-typing. These cells lines come under five major molecular subtypes namely the luminal A and B, HER-2+, triple- A and B subtypes. These cell lines have been well characterized and were utilized for understanding various aspects of breast cancers. Also, apart from providing an understanding of the molecular mechanisms associated with breast cancers, these cell lines have contributed significantly to areas such as drug testing. We present in this review the features of these cell lines, the studies conducted using them and the outcome of such studies. Also, the details about the culture conditions and study outcomes of the cell lines grown in 3-dimensional (3D) systems are presented.

Prophasing interphase chromatin for assessing genetic damages-The evolution, applications and the future prospects.

Premature chromosome condensation (PCC) involves induction of near-chromosome-like morphology to interphase chromatin. Experimental induction of PCC was achieved by somatic cell hybridization (SCH), an approach which evolved into a chemical-induction process. PCC presents most probably the only way in which cytogenetic assessment of damages can be analyzed in special situations such as availability of limited numbers of sample cells and for cells which have lost their ability to divide. Initial experiments on PCC were reported in late 1960s and the technique has evolved into one with wide range of applications owing to its increased efficiency in detecting primary DNA damages. Biodosimetry remains as the primary area which utilizes PCC technique to the maximum efficiency with several multiple-groups participating in collaborative exercises for biodosimetric applications. However, in spite of the advantages that the technique offers, it is yet to reach its full potential. This is due to the inherent limitations of the manner in which PCC is induced currently; by the somatic cell hybridization and chemical-induction processes. An approach which combines these two would sure help in taking PCC to its highest potential as the preferred technique for assessment of primary DNA damages. We present the chronological events of evolution of the PCC technique along with its applications. Also, the limitations of the technique along with the suggestions for further refinement of the PCC technique are discussed.

Agarose hydrogel induced MCF-7 and BMG-1 cell line progressive 3D and 3D revert cultures.

3D culture systems have enhanced the utility of cancer cell lines as they are considered closer to the in vivo systems. A variety of changes are induced in cells cultured in 3D systems; an apparent and striking feature being the spontaneous acquisition of distinct morphological entities. 3D reverts (3DRs) can be obtained by introducing 3D aggregates in scaffold/matrix-free culture units. It could be seen that the two cell lines used in this study exhibited differences in 3DR structures, though both were cultured on agarose hydrogels. Also, differences in 3DR formation, growth and survival were different. While 3D aggregates of several cell lines have been reported for a variety of studies, there are no studies that describe or utilize 3DRs. 3DRs can provide insights into complex events that can occur in cancer cells; especially as material to study metastasis, migration, and invasion.

Applications of Three-Dimensional Cell Cultures in the Early Stages of Drug Discovery, Focusing on Gene Expressions, Drug Metabolism, and Susceptibility.

Three-dimensional (3D) cell cultures represent advances in the way in which cells are grown in vitro, and they are gaining ground for a variety of applications. Advantages of 3D systems include reliable and realistic study endpoints. Cells grown in 3D conditions mimic in vivo conditions to an extent that is not possible with traditional 2D cell-culture methods. One major area in which 3D cell cultures have proven to make a difference is with drug discovery. The facets taken advantage of for drug discovery are the cells' complex microenvironment, including the production of extracellular matrices, altered gene expressions and their protein products, drug metabolism, and susceptibility to drugs when compared to 2D cultures. Altered protein and gene expressions are useful for drug-target identification. The susceptibility of 3D cultured cells for drug-induced effects can be used to obtain realistic data in a dose-dependent manner that can aid in identifying effective drug dosage. This review presents the contributions of 3D cell cultures for drug discovery, taking into consideration gene and protein expressions, drug metabolism, and susceptibility to drugs as the major focus points.

Human Brain Malignant Glioma (BMG-1) 3D Aggregate Morphology and Screening for Cytotoxicity and Anti-Proliferative Effects.

Two interesting aspects of cell lines grown in 3 Dimensional (3D) conditions are their distinct morphology and production of extracellular matrix (ECM). Also, it is known that 3D aggregates have different susceptibilities to damage-inducing agents compared to their 2D monolayer counterparts. We describe the effect of ECM on 3D aggregate morphology, the effect of cisplatin, bleomycin, and UV on the 3D aggregates and 2 Dimensional (2D) monolayers of the BMG-1 cell line. We also present a rapid method for analyzing cytotoxicity and anti-proliferative effects of 3D aggregates in 96-well plates. We utilized a single-step protocol using the dye resazurin. BMG-1 cells formed floating aggregates on 1% agarose hydrogels. The extent of ECM formed by them was dependent on number of cells seeded irrespective of the seeding density, which in turn directed the 3D aggregate compactness. The 3D aggregates were less susceptible to cisplatin and UV-induced cytotoxicity compared to 2D counterparts. The IC50 value of cisplatin was elevated at 210 µg/ml for the aggregates compared to 170 µg/ml for the monolayers. Exposure to UV for 0, 10, 20, and 30 min gave inhibition values of 2.98%, 8%, 22.99%, and 31.8% for the aggregates as compared to 3.06%, 7.5%, 39.4%, and 46.7% for the monolayers. While bleomycin-induced effects were unapparent when analyzed by vital staining for the doses used, the rapid, single-step method in 96-well plates was able to provide a dose-response for cytotoxicity and anti-proliferative effects. Also, comparative analysis of results obtained from vital staining and the single-step method demonstrates the reliability of the assay described. J. Cell. Physiol. 232: 685-690, 2017. © 2016 Wiley Periodicals, Inc.