Changes In Normal Cells Research Articles

Gambaran Kualitas Hidup Pada Pasien Ca Mamae Yang Menjalani Kemoterapi Di Rsup Dr. M. Djamil Padang

Cancer is a disease characterized by the presence of changes in normal cells into abnormal cells that are not controlled and can metastasize, both invading nearby tissues and distant biological tissues. According to estimates from the World Health Organization (WHO) in 2019, cancer was the first or second cause of death before the age of 70 in 112 out of 183 countries. This study aims to determine the picture of quality of life in ca mamae sufferers undergoing chemotherapy at RSUP dr. M.djamil padang. This type of research uses a descriptive approach. The sample was 60 respondents with a non-random sampling technique with accidental sampling technique. The measuring instrument used is in the form of a HARS (Hamilton Anxiety Rating Scale) questionnaire consisting of 14 pernyatann. The results of the study found that most of the breast cancer patients, namely as many as 78.3% who underwent chemotherapy in the chemotherapy room of RSUP M.Djamil, Padang City, had a poor quality of life. The conclusion of this study is that most of the quality of life of ca mamae patients undergoing chemotherapy is of poor quality. Advice for families to provide attention and support to people with ca mamae so that they can live a quality life because it can have an impact on the quality of health.

Cancer cells same as zombies reprogram normal cells via the secreted microenvironment.

The cancer microenvironment plays a crucial role in promoting metastasis and malignancy even in normal cells. In the present study, the effect of acidic and conditioned media of cancer cells (MDA-MB-231), separately and in combination, was studied for the first time on the cell death mechanisms and DNA methylation of normal fibroblasts (NIH/3T3). Cell survival of conditioned media was rescued by the addition of acidic media to conditioned media, as shown by the results. Cell metabolic activity is deviated in a direction other than the Krebs cycle by acidic media The mitochondrial metabolic activity of all groups was enhanced over time, except for acidic media. Unlike the highest amount of ROS in conditioned media, its level decreased to the level of acidic media in the combination group. Furthermore, cells were deviated towards autophagy, rather than apoptosis, by the addition of acidic media to the conditioned media, unlike the conditioned media. Global DNA methylation analysis revealed significantly higher DNA hypomethylation in acidic media than in normal and combination media. Not only were cells treated with conditioned media rescued by acidic media, but also DNA hypomethylation and apoptosis in the combination group were decreased through epigenetic modifications. The acidic and conditioned media produced by cancer cells can remotely activate malignant signaling pathways, much like zombies, which can cause metabolic and epigenetic changes in normal cells.

PROSPECTS OF THE USE OF RADIOPROTECTORS TO OVERCOME MITOCHONDRIAL DYSFUNCTION OF HEALTHY CELLS SURROUNDING THE IRRADIATED TUMOR

Radiation-induced changes in normal cells, including their mitochondria, from around the tumor can lead to the development of remote complications that negatively affect the effectiveness of radiation therapy. This determines the search for radioprotectors capable of overcoming the membrane barrier of mitochondria and suppressing their acquired dysfunction for selective protection of healthy tissues. The use of amifostine and melatonin radiomitigators, whose action is characterized by low toxicity, the ability to overcome acquired mitochondrial dysfunction, and unimpeded penetration into the mitochondria of various types of cells, is recognized as a reliable and effective means of protection. The appointment of the specified drugs to accompany radiation therapy of oncological patients is substantiated and recommended.

Cyto-toxic Effect of Nano Ceria bio fabricated with DNA

Cancer is a condition arising from the deregulation of several genes which causes change in normal cells to cancerous ones. The objective of all cancer treatment research is to eradicate cancer with little effect on normal cells. Nanostructured ceria of very small dimension (≈5nm) synthesized by co-precipitation method using deoxyribo nucleic acid (DNA) as capping agent has shown a promising antioxidant activity over almost all free radicals. Excessive reactive oxygen species have significant influence in the origination and development of numerous ailments, including cancer. So, the cyto-toxic effect of synthesized nanoceria on normal cells as well as on colon cancer cells is investigated. Inverted phase contrast microscope succeeded by MTT assay method were used to observe the survivability of cells. The investigation reveals a significant decline in the viability of cells when nanoceria, in the concentration range of 6.25–100μg/ml, was used on human cancer cells for 24,48 and 72 hours. The results showed that the activity of nanostructured ceria significantly depends on dose and time. For normal cells the reduction is less i.e., more than 75% of the cells are viable even if the concentration increases to 100μg/ml. But for cancer cells the percentage of viability decreases. Our study come up with a result that DNA assisted synthesized nanoceria has considerable cytotoxic effect, which seems likely to be effective as an anticancer agent while protecting healthy tissues.

Contribution of Tumor-Derived Extracellular Vesicles to Malignant Transformation of Normal Cells.

Tumor-cell-derived extracellular vesicles (EVs) are known to carry biologically active molecules of parental cells, which can actively modulate the tumor microenvironment. EVs produced by tumor cells play significant roles in the development and maintenance of tumor growth, metastasis, immune escape, and other important processes. However, the ability of EVs to induce the transformation of normal cells has hardly been investigated. This review discusses studies that describe the ability of tumor-cell-derived EVs to alter the metabolism and morphology of normal cells, causing changes associated with malignant transformation. Additionally, the horizontal transfer of oncogenes through EVs of tumor cells and the induction of epigenetic changes in normal cells, which leads to genomic instability and subsequent oncogenic transformation of normal cells, are also discussed.

Suppressive effects of dental pulp stem cells and its conditioned medium on development and migration of colorectal cancer cells through MAPKinase pathways.

Objective(s):Mesenchymal stem cells (MSCs) extensively interact with cancer cells and other stromal cells in the tumor microenvironment. However, the role of MSCs in colorectal cancer (CRC) development and metastasis is controversial. Strong evidence demonstrated that conditioned medium (CM) obtained from MSCs regulates main cellular functions such as proliferation, differentiation, migration, and communication due to its cell secretomes. This study was designed to determine the inhibitory effect of dental pulp stem cells (DPSC) and its extracted conditioned medium (DPSC-CM) in CRC progression.Materials and Methods:The inhibitory effects of DPSC-CM on growth, apoptosis, and migration of CRC cells were evaluated by resazurin, flow cytometry of propidium iodide (PI) stained cells, and wound closure assay, respectively. Western blotting detected the expression of MAPKinase and apoptotic proteins. Also, the homing ability of DPSCs and the invasion ability of CRC cells under indirect co-culture were assayed by the Boyden chamber assay. Results:DPSC-CM reduced the viability and induced the apoptosis of CRC cells significantly. Western blot analysis confirmed the increase in cytochrome C, phospho-JNK/SAPK to JNK/SAPK ratio, cleaved-caspase 8 and 3 in treated CRC cells with DPSC-CM, and decrease in phospho-ERK (P44/42 MAPK) to ERK (P44/42 MAPK) ratio, which are involved in induction of apoptosis and growth inhibition of cancer cells with minimal change in normal cells. Also, DPSCs could migrate (homing ability) to Caco2 and SW48 cells significantly.Conclusion:To sum up, DPSC-CM had significant apoptotic and growth inhibitory effects on the CRC cells through the MAPKinase and apoptosis signaling pathways.

X-rays induced IL-8 production in lung cancer cells via p38/MAPK and NF-κB pathway

Purpose It is reported inflammatory cytokine interleukin-8 (IL-8) could predict radiation-induced lung toxicity (RILT). RILT is believed to be a consequence of a cascade of cytokine production. It is considered that vascular endothelial cell and macrophages are the mainly source of cytokines. This study was investigated the production of IL-8 from cancer cells induced by X-rays may involve in the radiation-induced inflammation. Materials and methods We analyzed IL-8 in human lung cancer cell lines after expose to X-rays, and we also detect IL-8 in HUVEC cells and THP1 cells as endothelial cell and macrophage model to identify the change in normal cells after expose. Furthermore, we added the inhibitors to the culture with or without radiation to identify the role of MAPK and NF-κB pathways on the radiation-induced secretion of IL-8. Results Radiation could induce IL-8 production both in non-lung cancer cells (HUVECs and THP1 cells) and in lung cancer cells (A549 cells, H446 cells, PC-9 cells). Simultaneously, radiation activated p38/MAPK and NF-κB signal pathways in lung cancer cells. Moreover, p38/MAPK inhibitor SB203580 and NF-κB inhibitor BAY11-7082 could block the IL-8 up-regulated by X-rays but JNK inhibitor SP600125, ERK inhibitor U0126, ROS Scavenger NAC could not inhibit this phenomenon. Conclusions X-rays could induce IL-8 production in lung cancer cells, which may be related to the activation of p38/MAPK and NF-κB signaling pathway, providing a new point for elucidating the mechanism of radiation pneumonitis.

Inflammatory markers in cancer: Potential resources.

Cancer is a leading cause of death worldwide and a major burden on developing and less developed countries of the world with limited resources for prevention and effective treatment of cancer. Although cancer is multifactorial in origin, various epidemiological and experimental studies suggest that chronic inflammation has an important role in all stages of cancer, from initiation to progression and even survival of the patient. Inflammatory products like cytokines, chemokines, leucocytes, prostaglandins, cyclooxygenase, reactive oxygen and nitrogen species, metalloproteinase induce genetic and epigenetic changes in normal cells damaging its DNA, inhibiting its repair, altering transcription factors, preventing apoptosis, and stimulating angiogenesis, and thus resulting in carcinogenesis. Thus, these inflammatory mediators have a potential role to become cancer biomarkers for all stages of cancer as many of them can be measured in a cost-effective manner. However, large scale prospective trials are required to validate these potential cancer biomarkers. Nonetheless, a transition from potential to practical utilization of these markers will be an effective tool for the amelioration of cancer burden and mortality in a resource limited setting.

Abstract 1040: Characterization of extracellular vesicles in pancreatic cancer: Changing the way we think about tumorigenesis

Abstract Introduction: Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related deaths by 2030. Patients diagnosed with PDAC have a 5-year survival rate ~9%. Detection of pre-neoplastic lesions has the potential to improve treatment and overall survival in pancreatic cancer (PC) by 4-5 fold. However, there is currently no validated screening test for the non-invasive and accurate detection of early stage PC. Extracellular vesicles (EVs) arise from invagination of, and secretion from, cells of all tissue types. Their role in mediating PC tumorigenesis however, is just beginning to be studied. We have found that EVs derived from PC cells (cancer-derived EVs, CDEVs), can impart significant phenotypic and metabolic transformational effects on normal human pancreas cells (hTERT-HPNE and H6c7), tantamount to an epithelial-to-mesenchymal transition (EMT). The goal of this study was to characterize the biochemical cargo of EVs toward understanding their role in PC tumorigenesis. Results: We isolated CDEVs from various established and PDX PC cell lines. After validating enrichment of EV fractions with quantitative ELISA, immunoblot and transmission electron microscopy (TEM), we added CDEVs to culture media of normal cells. We found that CDEVs induce a myriad of changes in normal cells, including significant morphological changes, increased proliferation and an uncharacteristic invasive capability. CDEVs also caused a bioenergetic switch in normal cells, from a quiescent, aerobic profile to a highly energetic and glycolytic profile. Comparative analyses of CDEVs and normal-derived EVs (NDEVs) using 1H- and 13C-NMR showed significantly different biochemical fingerprints. Interestingly, we found that NDEVs contained more amino acids than CDEVs, while CDEVs contained elevated amounts of urea and lactic acid. We next designed a 13C-isotope labeling flux experiment to trace the fate of carbon metabolized from 13C-glucose fed to cells in culture. Using UHPLC/Q-ToF-MS, we identified several differentially labeled isotopes in CDEVs, particularly surrounding lipid biosynthesis and folate. Conclusions: Our results indicate that CDEVs confer enormous transformational properties to normal human pancreas cells in vitro. We hypothesize that EVs could impart tumorigenic properties to normal cells in vivo and this influence could unveil novel mechanisms underpinning cancer onset and progression. We show that there are robust biochemical differences between CDEVs and NDEVs. Biochemical characterization of EV cargo can uncover abrogated pathways in which signaling is mediated by EVs. These signals may be detectable before progression of PC to PDAC, leading to the development of assays for earlier diagnosis in patients. Citation Format: Charles P. Hinzman, Michael Girgis, Yaoxiang Li, Meth Jayatilake, Meena Rajagopal, Partha P. Banerjee, Amrita K. Cheema. Characterization of extracellular vesicles in pancreatic cancer: Changing the way we think about tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1040.

Cytotoxic effects of juglone and Pterocarya fraxinifolia on prostate cancer cells

Aim:Juglone with naphthoquinone structure has medicinal properties and its anticarcinogenic and antioxidant effects have been proven. In this research, the cytotoxic and apoptosis effects of juglone and Pterocarya fraxinifolia (PF) methanolic extract on human prostate cancer cells were studied.Materials and Methods:The PC3 and DU145 human cancer cells and normal cells of primary prostate epithelial cells (ATCC PCS-440-010) were treated with juglone and PF extract at the concentrations of 10, 50, 100, 500, and 1000 μg/mL for 24, 48, 72, and 96h. The morphological changes were examined by reversed microscope. The survival percentage of cell lines was evaluated by MTT (3,4,5-dimethylthiazole-2-yl-2,5-diphenyltetrazolium bromide) test. The rate of apoptosis and expression of AR and CLU genes were examined by flow cytometry and real-time polymerase chain reaction.Results:All concentrations after 24h caused morphological changes in PC3 and DU145 cells, and these changes were intensified after 48, 72, and 96h. Also, concentrations of 100 and 500 μg/mL caused morphological changes in normal cells. The results of MTT test showed a significant decrease in PC3 and DU145 cell survival rate at 50, 100, and 500 μg/mL concentrations (P < 0.05). Juglone at 10 μg/mL concentration induced apoptosis in cancer cell lines.Conclusion:Juglone and PF could decrease the growth of cancer cell lines through the mitochondrial pathway. So PF could be considered as a potential candidate for therapeutic herbal medicine in treating cancers.

P2.11-14 Malignancy Associated Change and The LuCED® Test for Detection of Early Stage Lung Cancer

Early detection remains the most reliable and effective strategy for curing lung cancer. Many approaches, however, are limited by poor sensitivity or specificity that increase health care costs and potentially risk patient health through unneeded procedures. The association between cell morphology and cancer has been established in the pathology literature. However, through the so-called field effect, cancer can introduce subtle morphological changes into non-cancerous cells that are proximal to the tumor site. The Cell-CT™ platform and LuCED® test represent a promising new method for detecting lung cancer with high (92%) sensitivity and (95%) specificity based on cytologically abnormal cells. In this research we investigate use of the Cell-CT to detect malignancy associated changes in normal cells near the cancer with a view towards enhancing LuCED test performance. We present a study of 3D morphological alterations in non-cancer cells obtained from sputum of healthy subjects and biopsy confirmed lung cancer patients. Three major cell types were analyzed from 235 patients: bronchial epithelial columnar, squamous intermediate, and mature macrophages. We used the Cell-CT™ platform to measure over 700 different structural biomarkers for each cell. The measurements were used to define prominent clusters of cells through a hierarchical process that were then used under supervised learning, with case status as ground truth, to create classifiers that optimally separated cells from cancer vs. normal cases. The table gives classifier development and performance characteristics:Tabled 1Cell TypeNumber of cells from cancer patientsNumber of cells from normal patientsCluster-based Supervised Learning – aROCSquamous Intermediate23166840.94Macrophages496050400.94Columnar cells322732340.92 Open table in a new tab Our results indicate that the Cell-CT can discriminate cell features that are too subtle to distinguish by a human. The study suggests that detection of cells with Malignancy Associated Changes may be used to further enhance the LuCED test’s performance beyond published levels.

Sodium bicarbonate cotransporter NBCe1 affects the growth and motility of prostate cancer cell lines LNCaP and PC3

Prostate cancer is the most common cancer in men. Prostate cancer develops mainly in older men: 6 cases in 10 are diagnosed in men aged 65 or older while it is rare before age 40, according to the report by the American Cancer Society. The average age at the time of diagnosis is ~66. Similar to other cancers, prostate cancer is developed from genetic and epigenetic changes in normal cells, and environmental and systemic stresses also make significant contributions to cancer aggravation. One of such stresses is acidic tumor microenvironments, which is caused by excessive glycolytic cancer cell metabolism, hypoxia, and inefficient blood perfusion. Acidic pH has pleiotropic effects on cancer cell proliferation, clonal evolution/selection and metastasis, and also modulates immune cell functions. The processes of the pH gradient in cancer cells have recently become targets for anti‐cancer therapies.In this study, we examined the effects of acidic pH environments on prostate cancer growth. We focuses on the Na/HCO3 cotransporter NBCe1 (SLC4A4) that moves Na+ and HCO3− into cells and buffers intracellular H+, resulting in extracellular acidification. To examine whether NBCe1 activity is stimulated in proliferating cancer cell lines, we incubated human prostate cancer cell lines LNCaP and PC‐3 cells and human breast cancer cell line MCF7 cells in hypoxic (1% O2, 5% CO2) conditions for 3 days and measured pHi using the fluorescent pH‐sensitive dye BCECF‐AM, and fluorescent images (excitation at 490 and 440 nm; emission at 535 nm) were captured at different time intervals. pH values were generated by the high K+‐nigericin calibration. The solutions contained 100 μM EIPA to inhibit endogenous NHE. After a rapid initial CO2‐induced acidification, the pHi was recovered. The recovery was significantly increased in hypoxic conditions. It is also inhibited by DIDS and S0859, which block most NBCs. In RT‐PCR with the primers that amplify NBCe1 transcripts, the amplification products with the expected size were detected in both prostate cancer cells and breast cancer cells. Next, we performed immunoblot to examine NBCe1 protein expression in cells. Determined by densitometric quantitation of immunoreactive bands and normalization of pixel intensities to β‐actin, the NBCe1 expression in LNCap and PC‐3 cells was increased in hypoxic conditions, whereas the expression in MCF7 remained unaffected. The NBCe1 upregulation in prostate cancer cells was small, but statistically significant (p &lt; 0.05; paired, two‐tailed Student's test; n = 4). To test whether pH changes mediated by the Na/HCO3 transporter affects the growth of prostate cancer cells, we treated cells with the blocker N‐cyanosulphonamide S0859 (50 μM) in normoxic and hypoxic incubations and counted cells 5 days later using a propidium iodide staining protocol. S0859 reduced the number of cells that were proliferated under the hypoxic condition (p&lt;0.05, n=6; unpaired two‐tailed Student t‐test). We also treated cells with higher amounts of the drug and observed more profound inhibition on cell growth (data not shown). These results demonstrate that NBCe1 is the key protein responsible for acidic microenvironments in human prostate cancer.Support or Funding InformationSupported by the Emory Winship Cancer Center Pilot GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Abstract 2408: Epigenomic characterization of MSC from myeloid malignancies

Abstract Ineffective hematopoiesis is a hallmark of myeloid malignancies such as Myelodysplastic Syndromes (MDS) or Acute Myeloid Leukemia (AML). Our previous research has shown that mesenchymal stromal cells (MSC), one of the main cellular regulatory components of the bone marrow niche and essential for the differentiation of new blood cells from Hematopoietic Stem Cells (HSC), are molecularly and functionally impaired in MDS and AML patients. More specifically, these cells display reduced proliferative and differentiation capacities, an increased cellular senescence, or the altered expression of key hematopoietic factors, defects that can provide an explanation for the cytopenia associated with these diseases. But most interestingly, we have also discovered that MDS- and AML-derived MSC present aberrant methylomes. Our data, based on 450k DNA methylation microarrays, show that these abnormalities are shared by the MSC from both malignancies and affect key transcription factors like TBX15, PITX2 or HOXB6, formerly linked to differentiation and development. This results can account for some of the defects observed in the disease-related MSC and represent an important paradigm for cancer-induced epigenetic changes in normal cells. Aiming to further explore the importance of this epigenetic deregulation, we performed RNAseq experiments using MSC from healthy donors as well as from MDS and AML patients. This approach not only suggests a profound impact of the epigenetic aberrancies on gene expression, but also allowed us to identify TGFβ as the soluble factor that, released by the malignant cells in the bone marrow, appears to trigger the above-mentioned defects in the MDS- and AML-derived MSC. Indeed, our results show that, when treated with TGFβ, healthy MSC undergo the deregulation of TBX15, PITX2 or HOXB6 and develop some of the observed aberrant phenotypes. Conversely, the treatment of MDS- and AML-derived MSC with a TGFβ antagonist like SD208 can abrogate these effects. Finally, our preliminary DNA methylation data reveal a very similar deregulation in the methylome of MSC from Multiple Myeloma (MM) patients, strongly suggesting that the same scenario could also be common to other hematological malignancies from the lymphoid branch. Citation Format: Manuel Rodríguez-Paredes, Stefanie Geyh, Mahshid Gazorpak, Julian Gutekunst, Felix Bormann, Rainer Haas, Thomas Schröder, Frank Lyko. Epigenomic characterization of MSC from myeloid malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2408. doi:10.1158/1538-7445.AM2017-2408

Is Shape of Cancer Cell Correlated with its Invasiveness?

This work is based on the hypothesis that shape of the cell on substrate is determined by the active mechanical properties of the cytoskeleton. Since invasive cancer cells are believed to have altered mechanical properties compared with non-invasive cells, it follows that changes in shape of cancer cells may correlate with the acquisition of invasive capacity. We try to understand whether there is a correlation between cancer cell shape and its phenotypic state. To proceed, we first need to capture cell shape and quantify it. Cells were cultured on glass substrates and then were fixed and stained for actin cytoskeleton. Images were processed to eliminate background noise. In order to quantitatively describe cell's shape, we chose a set of Zernike moments, which are widely used in image recognition and are calculated from a set of orthogonal basis function. Since we are dealing with high dimensional data we used principal component analysis (PCA) to examine changes in cell shape. We also used a machine learning neural network classifier to distinguish between different cell populations. We use paired osteosarcoma cancer lines which consist of a parental line that rarely undergoes metastasis and a line derived from parental line that almost always does. Comparing shape changes of invasive cancer line versus their paired low invasive cancer line gives insight to correlation between cell shape and invasiveness. To understand cytoskeletal mechanism behind this relation we perturbed cytoskeleton with different pharmaceutical drugs which each targets different element of the cytoskeleton and looked at the patterns of drug effect on cell shape. We also characterize shape changes of normal cells during their genetic step-wise transformation to cancer cells. [1]S. M. Lyons et. al. Biology Open 2016, 5, 289-299. [2]E.Alizadeh et. al. Integrative Biology, 2016, http://dx.doi.org/10.1039/C6IB00100A.

Leveraging premalignant biology for immune-based cancer prevention

Prevention is an essential component of cancer eradication. Next-generation sequencing of cancer genomes and epigenomes has defined large numbers of driver mutations and molecular subgroups, leading to therapeutic advances. By comparison, there is a relative paucity of such knowledge in premalignant neoplasia, which inherently limits the potential to develop precision prevention strategies. Studies on the interplay between germ-line and somatic events have elucidated genetic processes underlying premalignant progression and preventive targets. Emerging data hint at the immune system’s ability to intercept premalignancy and prevent cancer. Genetically engineered mouse models have identified mechanisms by which genetic drivers and other somatic alterations recruit inflammatory cells and induce changes in normal cells to create and interact with the premalignant tumor microenvironment to promote oncogenesis and immune evasion. These studies are currently limited to only a few lesion types and patients. In this Perspective, we advocate a large-scale collaborative effort to systematically map the biology of premalignancy and the surrounding cellular response. By bringing together scientists from diverse disciplines (e.g., biochemistry, omics, and computational biology; microbiology, immunology, and medical genetics; engineering, imaging, and synthetic chemistry; and implementation science), we can drive a concerted effort focused on cancer vaccines to reprogram the immune response to prevent, detect, and reject premalignancy. Lynch syndrome, clonal hematopoiesis, and cervical intraepithelial neoplasia which also serve as models for inherited syndromes, blood, and viral premalignancies, are ideal scenarios in which to launch this initiative.

Abstract 2025: SIX1 overexpression extends the lifespan of normal human keratinocytes and promotes epithelial-mesenchymal transition

Abstract SIX1, a homeodomain-containing transcription factor, has a critical role in the expansion of progenitor cells during embryogenesis. The overexpression of SIX1 contributes to tumorigenesis, promoting malignant transformation and metastasis. Six1 promotes tumor progression by induction of epithelial-to-mesenchymal transition (EMT). In HPV16-transformed human keratinocytes (HKc), SIX1 overexpression produces EMT and a differentiation-resistant phenotype at early stages of in-vitro progression, and transforms cells to malignancy at late stages. We transfected normal HKc with a plasmid encoding human-SIX1 in order to determine to what extent SIX1 overexpression may induce growth, differentiation and EMT changes in normal cells. We also studied the effects of expressing the HRas-V12 oncogene while downregulating the p53 tumor suppressor gene in normal HKc expressing SIX1, in order to push these keratinocytes to a precancerous state. Normal HKc overexpressing SIX1 (HKc/SIX1) or a combination of SIX1, HRas-V12 and p53i (HKc/ALL3) grow continuously while their vector-transfected controls senesced. Continuous growth past the end of the normal lifespan was also observed in normal human fibroblasts overexpressing SIX1. HKc/SIX1 and HKc/ALL3 exhibit a spindle-shape and fibroblastic appearance rather than the cuboidal morphology of epithelial cells. They are also morphologically distinct from SIX1-overexpressing human fibroblasts. HKc /SIX1 and HKc/ALL3 grow best in KSFM supplemented with 10% FBS and 1mM CaCl2, while normal HKc grow best in KSFM with 0.1 mM CaCl2. RT-PCR confirmed that mRNA levels of SIX1 were significantly increased and p53 levels were significantly decreased, and that HRasV12 was expressed in HKc/ALL3. The growth rates of HKc/SIX1 and HKc/ALL3 are comparable to those of early-stage HPV16-immoratlized cells (HKc/HPV16). Invasion assays demonstrated that HKc/SIX1 are more invasive than normal HKc, and their invasiveness is comparable to that of HKc/HPV16. The invasiveness of HKc/ALL3 is greater than that of late-stage, differentiation-resistant HPV-immortalized cells (HKc/DR), which are about 2.5-fold more invasive than HKc/HPV16. In conclusion, SIX1 promotes continued proliferation and invasiveness, signs indicative of cellular transformation, even in the background of normal HKc. Currently, we are exploring the gene expression changes induced by SIX1 in normal cells. Additionally, we plan to investigate whether these cells express stem cell markers. The results of these studies suggest that SIX1 facilitates the establishment of primary human keratinocytes into immortal cell lines, or perhaps into stem-like cells with extended proliferation potential. Citation Format: Maria Hosseinipour, Aspasia Amiridis, Diego Altomare, Kim E. Creek, Lucia Pirisi. SIX1 overexpression extends the lifespan of normal human keratinocytes and promotes epithelial-mesenchymal transition. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2025.

Epigenetic drift, epigenetic clocks and cancer risk.

It is well-established that the DNA methylation landscape of normal cells undergoes a gradual modification with age, termed as 'epigenetic drift'. Here, we review the current state of knowledge of epigenetic drift and its potential role in cancer etiology. We propose a new terminology to help distinguish the different components of epigenetic drift, with the aim of clarifying the role of the epigenetic clock, mitotic clocks and active changes, which accumulate in response to environmental disease risk factors. We further highlight the growing evidence that epigenetic changes associated with cancer risk factors may play an important causal role in cancer development, and that monitoring these molecular changes in normal cells may offer novel risk prediction and disease prevention strategies.

Mitochondrial Alteration: A Major Player in Carcinogenesis

In recent years we have witnessed mounting evidences on how a normal cell changes its cellular signaling and metabolic pathways to become highly proliferative cancer cell. Since mitochondrion is a major hub of energy production and several metabolic pathways, it is taking the center stage in defining alterations in energy homeostasis and metabolic rerouting of cancer cell proliferation. Similarly, mutations in the mitochondrial genome in cancer are providing new insights on how these mutations affect mitochondrial functions and change the oncogenic signaling and apoptosis mechanism. In this review, we will summarize these important mitochondrial mechanisms that contribute significantly in the progression of cancer. Further therapeutic approaches targeted to these altered mitochondrial mechanism in cancer are discussed. This review is a part of special issue on Mitochondria: implications in human health and diseases.

CTCFL (BORIS) mRNA Expression in a Peripheral Giant Cell Granuloma of the Oral Cavity.

Peripheral giant cell granuloma (PGCG) is a relatively common benign reactive lesion of the oral cavity which can occur at any age. CTCFL/BORIS (CTCF like/Brother of the Regulator of Imprinted Sites) and CTCF (CCCTC-binding factor) are paralogous genes with an important role in the regulation of gene expression, genomic imprinting, and nuclear chromatin insulators regulation. BORIS expression promotes cell immortalization and growth while CTCF has tumor suppressor activity; the expression pattern may reflect the reverse transcription silencing of BORIS. The aim of this work was to describe a histopathological and molecular approach of an 8-year-old pediatric male patient with PGCG diagnosis. It was observed that the PGCG under study expressed CTCF as well as BORIS mRNAs alongside with the housekeeping gene GAPDH, which may be related to possible genetic and epigenetic changes in normal cells of oral cavity.

Comparative evaluation of antidiabetic activity of Rosmarinus officinalis L. and Chamomile recutita in streptozotocin induced diabetic rats

Herbal formulations are getting more importance in the treatment of diabetes, cancer and hepatic disorder because of the hazardous adverse effects of the current therapy. Especially diabetes can be controlled by Allopathic medicine as well as Herbal medicine. A comparison was made between the antidiabetic activities of water extracts of leaves of Rosmarinus Officinalist and Chamomile recutitain Streptozotocin- induced diabetic rats. Serum Fasting blood glucose and other biochemical parameters such as total protein; carbohydrate hydrolyzing enzyme; α – amylase, total antioxidant capacity, liversteoteosis; total cholesterol, Triacylglycerol, were determined at the dose 200 mg/kg body weight for 21 days. There is a significant increase decrease in serum fasting blood glucose (p<0.001) , increase in body weight and improvement in the other biochemical parameters with treatment of herbal formulations which altered in diabetic rats There was a significant decrease in serum glucose level (p<0.001), increase in body weight and changes in normal cells was observed with treatment of the above mentioned extracts which altered in diabetic rats as compared to the control rats standard.