Nucleus Of Cancer Cells Research Articles

Automated Detection Model in Classification of B-Lymphoblast Cells from Normal B-Lymphoid Precursors in Blood Smear Microscopic Images Based on the Majority Voting Technique

Introduction. Acute lymphoblastic leukemia (ALL) is the most common type of leukemia, a deadly white blood cell disease that impacts the human bone marrow. ALL detection in its early stages has always been riddled with complexity and difficulty. Peripheral blood smear (PBS) examination, a common method applied at the outset of ALL diagnosis, is a time-consuming and tedious process that largely depends on the specialist’s experience. Materials and Methods. Herein, a fast, efficient, and comprehensive model based on deep learning (DL) was proposed by implementing eight well-known convolutional neural network (CNN) models for feature extraction on all images and classification of B-ALL lymphoblast and normal cells. After evaluating their performance, four best-performing CNN models were selected to compose an ensemble classifier by combining each classifier’s pretrained model capabilities. Results. Due to the close similarity of the nuclei of cancerous and normal cells, CNN models alone had low sensitivity and poor performance in diagnosing these two classes. The proposed model based on the majority voting technique was adopted to combine the CNN models. The resulting model achieved a sensitivity of 99.4, specificity of 96.7, AUC of 98.3, and accuracy of 98.5. Conclusion. In classifying cancerous blood cells from normal cells, the proposed method can achieve high accuracy without the operator’s intervention in cell feature determination. It can thus be recommended as an extraordinary tool for the analysis of blood samples in digital laboratory equipment to assist laboratory specialists.

The variation of 5 -carboxycytosine localization within the nucleus of normal and cancerous cells

The variation of 5 -carboxycytosine localization within the nucleus of normal and cancerous cells

Carbon Dot Nanoarchitectonics: Quadruple Conjugated Nano Model for Superior Cancer Cell-Nucleus Targeting and Multi Drug Delivery

Carbon Dot Nanoarchitectonics: Quadruple Conjugated Nano Model for Superior Cancer Cell-Nucleus Targeting and Multi Drug Delivery

Evaluation of Anticancer Effect of Lippia nodiflora Leaf Extract on Lung Cancer Cells

Background: Lung cancer, also known as bronchogenic carcinoma, is a kind of cancer that begins in the lung parenchyma or inside the bronchi. In the United States, it is one of the top causes of cancer-related fatalities. Lippia nodiflora is a flowering, broadleaf plant native to South America. Lippia nodiflora has been used as a natural medicine for a number of disorders due to its antioxidant, anti-inflammatory, anti-bacterial, and anti-tumor characteristics.
 Aim of the Study: To evaluate the anticancer effect of Lippia nodiflora leaf extract on lung cancer.
 Materials and methods: The cytotoxic effect of Lippia nodiflora leaf extract was assessed by an MTT assay. The A-549 cells were treated with different concentrations of Lippia nodiflora leaf extract (10-120 μg) for 24h. Analysis of changes in the cell morphology is examined by phase contrast microscope. For the nuclear morphological changes analysis, DAPI staining was used in lung cancer cells. The apoptotic nuclei were viewed under a fluorescent microscope.
 Results: In our study, Lippia nodiflora leaf extract treated cells showed significant reduction of cell viability in lung cancer cells. 50% of the inhibition was observed in a 20 μg/ml concentration, which has been taken as the inhibitory concentration (IC-50) which was used for further analyses. The pro-apoptotic effect of Lippia nodiflora leaf extract in A549 cell line was determined by DAPI staining under a fluorescence microscope. Lippia nodiflora leaf extract treatment alters the nuclear morphology and condensed nuclei in lung cancer cells at 20μg/ml concentration.
 Conclusion: Overall, it can be concluded that Lippia nodiflora presents anticancer activity at 20μg/ml concentration at 24 hrs againstthe lung cancer cell line. However, further research is warrented to find out the molecular mechanism of cytotoxicity of effect this plant extract against lung cancer.

Breast radiotherapy with kilovoltage photons and gold nanoparticles as radiosensitizer: An in vitro study.

PurposeWe investigated the dose enhancement and internalization of gold nanoparticles (AuNPs) used as a radiosensitizer agent for rotational radiotherapy of breast cancer using a kilovoltage (kV) X‐ray beam.MethodsHuman breast cancer cells MDA‐MB‐231 were incubated with or without 100 μg/mL (4.87 nM) or 200 μg/mL (9.74 nM) 15 nm AuNPs and irradiated with 100 kV, 190 kV, or 6 MV X‐rays. To assess the toxicity of the AuNPs, we performed a Sulforhodamine B assay. Using atomic absorption spectroscopy, scanning electron microscopy, transmission electron microscopy, and time‐lapse optical microscopy (rate of 2 frames per minute), we carried out a quantitative assessment of the amount of gold internalized by MDA‐MB‐231 cells and a characterization of the static and dynamical aspects of this internalization process.ResultsNo effect of AuNPs alone was shown on cell viability. Time‐lapse optical microscopy showed for the first time AuNPs cellular uptake and the dynamics of AuNPs internalization. Electron microscopy demonstrated AuNPs localization in endosomal vesicles, preferentially in the perinuclear region. After irradiation at doses up to 2 Gy, cell survival fraction curves showed increased mortality with AuNPs, with respect to irradiation without AuNPs. The highest effect of radioenhancement by AuNPs (at 9.74 nM AuNPs concentration) was observed at 190 kV showing a dose enhancement factor of 1.33 ± 0.06 (1.34 ± 0.02 at 100 kV), while at 6 MV it was 1.14 ± 0.06.ConclusionsThe observed radio‐sensitization effect is promising for future radio‐enhanced kV radiotherapy of breast cancer and quantitatively in the order of previous observations for 15 nm AuNPs. These results of a significant dose enhancement were obtained at 15 nm AuNPs concentration as low as several nanomolar units, at dose levels typical of a single dose fraction in a radiotherapy session. Dynamical behavior of the 3D spatial distribution of 15 nm AuNPs outside the nucleus of single breast cancer cell was observed, with possible implications for future models of AuNPs sensitization.

Spider Toxin Peptide-Induced NIR Gold Nanocluster Fabrication for GSH-Responsive Cancer Cell Imaging and Nuclei Translocation.

Goldnanoclusters (GNCs) have become a promising nanomaterial for bioimaging because of their unique optical properties and biocompatibility. In this study, lycosin-I peptide, which possesses a highly selective anticancer activity by affecting the permeability of cancer cell membrane, was firstly modified for constructing fluorescent GNCs (LGNCs) for bioimaging of tumor cells. The obtained LGNCs exhibited strong near-infrared (NIR) fluorescence, which can be further enhanced by the peptide-induced aggregation and selectively stained three cancerous cell lines over normal cell lines with low intrinsic toxicity. After uptake by tumor cells, LGNC aggregates can be depolymerized into ultrasmall nanoclusters by high-level glutathione (GSH) and realize the nuclear targeting translocation. Collectively, our work suggests the potential of natural active biomolecules in designing NIR fluorescent GNCs for bioimaging.

Overexpression of methyltransferase-like 3 and 14 in oral squamous cell carcinoma.

This study aimed to determine expressions of methyltransferase-like 3 (METTL3) and METTL14, two enzymes essential for mRNA methylation at the adenosine (m6 A), in oral squamous cell carcinoma (OSCC) and to investigate in vitro aggressiveness of their aberrant expressions. METTL3 and METTL14 expressions in 50 OSCC and 11 normal oral tissues were examined by immunohistochemistry. METTL3 and METTL14 expressions and m6 A amounts were determined in three OSCC cell lines, including HN5, HN6, and HN15. Cell proliferation, migration, and invasion were studied by BrdU, wound healing, and Transwell chamber assays, after silencing of METTL3, METTL14, or both by siRNA transfection. Immunostaining of METTL3 and METTL14 was localized in cancer cell nuclei. The mean percentages of METTL3- and METTL14-positive cells were significantly increased in OSCC tissues (p<0.001). The percentages of METTL3- and METTL14-positive cells were correlated with the advanced pTNM stages (p<0.05) and with the degrees of histopathological differentiation in OSCC (r=0.564 and r=0.316, respectively; p<0.001). By the COX multivariate analysis, both overexpressed METTL3 and METTL14 were significantly associated with short overall survival (p<0.05). Both METTL3 and METTL14 expressions and the m6 A amounts were significantly increased in HN6 (p<0.05). Silencing of METTL3 and METTL14 in HN6significantly inhibited cell proliferation (p<0.01), but it failed to mitigate cell migration or invasion. METTL3 and METTL14 are overexpressed in OSCC tissues and in the HN6 OSCC cell line that promotes cell proliferation. Overexpressed METTL3 or METTL14 is found to be an independent prognostic factor for short overall survival in patients with OSCC.

Local DNA microviscosity converts ruthenium polypyridyl complexes to ultrasensitive photosensitizers

In this manuscript, we have reported how DNA binding of ruthenium polypyridyl complexes (RPCs) make them more efficient photosensitizer. The DNA binding of RPCs results in a significant enhancement of their emission intensity and excited-state lifetime. As a result, upon visible-light irradiation, these complexes generate reactive singlet oxygen, which causes selective DNA damage and kills cancer cells. This investigation also demonstrates the effect of light-driven RPCs on bacterial growth arrest through DNA nicking and differential localization in cancer and non-cancer cells. The local DNA microviscosity suppresses the non-radiative pathways which causes a large enhancement in the emission intensity and the excited state lifetime. The visible-light-triggered singlet-oxygen efficiently produces nick in DNA and inhibits bacterial growth. RPCs also localize inside the cancer cell nucleus and in the vicinity of the nuclear membrane of non-cancerous cells, confirmed by live-cell confocal microscopy. The results provide a facile platform for the novel antibiotic intended discovery combined with cancer therapy.

Endocytic protein intersectin1-S shuttles into nucleus to suppress the DNA replication in breast cancer

Breast cancer is the most common type of cancer worldwide. However, the well-known molecular biomarkers are not enough to meet the needs of precision medicine. In search for novel targets in this regard, we reported ITSN1 (intersectin1) as one of the candidates through mRNA microarray analysis. In the present study, we reported that endocytic protein ITSN1-S exists not only in the cytoplasm but also in nuclei of breast cancer cells. ITSN1-S′ functional nuclear localization signal is within its residues 306–312. Its nuclear export signal (NES) resides within its SH3 domains. We also found, the interaction between the CC domain of nuclear ITSN1-S and the NT domain of nuclear DNA helicase II (NDH II) directly suppressed the DNA replication and nascent DNA synthesis by inhibiting the R-loops resolution in breast cancer cells. Furthermore, the interaction between the EH domains of cytoplasmic ITSN1-S and PI3KC2α inhibit cell migration and invasion by inactivating the PI3KC2α-AKT pathway. Our results were confirmed in both ITSN1 gene knockout cells and in vivo assays. Finally, our clinical data showed a potential application of the combined consideration of the cytoplasmic and nuclear ITSN1-S as an independent prognosis factor. In conclusion, our study revealed ITSN1-S′ novel positioning in the nuclei of breast cancer cells, its function in suppressing DNA replication, and its potential application in improved breast cancer prognosis.

P66.08 Differential Expression of INSM1 Between Pure SCLC and LCNEC After Surgical Resection and Its Clinicopathological Significance

According to WHO diagnostic criteria, both Small cell lung cancer (SCLC) and Large cell neuroendocrine (LCNEC) are categorized into high grade neuroendocrine carcinoma, based on morphological features and immunostaining of neuroendocrine markers including CD56, chromogranin A (CgA) and synaptophysin (Syn). However, the above markers have limited satisfactory significance of specificity and sensitivity. Insulinoma-associated protein 1 (INSM1) is highly expressed in SCLC cells as a zinc finger protein, and numerous studies have shown that INSM1 positive staining has a strong correlation with NE differentiation in SCLC and an alternative diagnostic biomarker for SCLC, but few studies are involved in LCNEC and its significance remains unclear in LCNEC. The aim of this study was to investigate the potential of INSM1 as a neuroendocrine marker by comparing the expression of INSM1 in SCLC and LCNEC and its clinicopathological significance. 347 surgically resected lung cancer including 290 pure SCLC and 57 pure LCNEC samples were retrieved in this retrospective study. INSM1 expression was detected by immunohistochemistry staining in SCLC and LCNEC tissue microarrays. The different expression of INSM1 between SCLC and LCNEC as well as the correlation with clinicopathological features including gender, age, smoking history, tumor location, AJCC 7th stage, reginal lymph node metastasis, pleural invasion and tumor thrombosis were analyzed by chi-square test. Of the included patients, male/female ratio was 2.33 (203/87) in SCLC and 10.4 (52/5) in LCNEC, the smoker rate was 64.5% (187/290) in SCLC and 43.9% (25/57) in NSCLC. According to the AJCC Cancer Staging Manual (7th edition), the SCLC patients were 29.0% (84/290) in stage I, 27.2% (79/290) in stage II, 40.7% (118/290) in stage III, and 3.1% (9/290) in stage IV, while the NSCLC patients were 49.1% (28/57) in stage I, 26.3% (15/57) in stage II, and 24.6% (14/57) in stage III, respectively. INSM1 was localized in the nucleus of lung cancer cells, a significant higher positive rate was detected in SCLC than that in LCNEC (271/290,93.4% in SCLC; 32/57,56.1% in LCNEC; p<0.05). INSM1 protein expression significantly correlated with smoking history (p=0.023), pleural invasion (p=0.010), and a positive trend of regional lymph node metastasis (p=0.052) in SCLC, but did not correlate with any of the clinicopathological features in LCNEC (p＞0.05). INSM1 protein was found no prognostic significance neither in SCLC nor in LCNEC (p＞0.05). INSM1 protein is highly expressed in SCLC and positively correlated with pleural invasion and smoking history. In contrast, INSM1 expression in LCNEC is significant lower and does not correlated with such clinicopathological features. Further research needs to be done on exploring the potential different underlying mechanisms and diagnostic significance of INSM1 in SCLC and LCNEC.

2-Methoxy-estradiol Loaded Alpha Lipoic Acid Nanoparticles Augment Cytotoxicity in MCF-7 Breast Cancer Cells.

The therapeutic effectiveness of anticancer drugs with a selective target for the nucleus of cancer cells may be improved by experimental approaches. In this regard, the formulation of anticancer drugs is considered one of the best ways to improve their effectiveness in targeting cancerous tissues. To enhance the anticancer activity of 2-methoxy-estradiol (2 ME) for breast cancer, 2-methoxyestradiol loaded alpha lipoic acid nanoparticles have been formulated. The prepared formula was observed to be spherical with a nanometer-scale and low PDI size (.234). The entrapment efficiency of the 2ME-ALA NPs was 87.32 ± 2.21% with > 85% release of 2 ME within 24 h. There was a 1.2-fold increase in apoptosis and a 3.46-fold increase in necrosis of the MCF-7 cells when incubated with 2ME-ALA NPs when compared to control cells. This increased apoptosis was also associated with increased ROS and increased p53 expression in 2ME-ALA NPs treated cells compared to the raw-2 ME group. Evaluation of cell-cycle data showed a substantial arrest of the G2-M phase of the MCF-7 cells when incubated with 2ME-ALA NPs. At the same time, a dramatically increased number of pre-G1 cells showed the increased apoptotic potential of the 2 ME when administered via the proposed formulation. In the end, the differential upregulation of caspase-3, p53, and ROS in MCF-7 cells established the superiority of the 2ME-ALA-Ms approach in targeting breast cancer. In summary, these results demonstrate that 2ME-ALA NPs are an efficient delivery tool for controlling the growth of breast cancer cells.

DNA Binding Mode Analysis of a Core-Extended Naphthalene Diimide as a Conformation-Sensitive Fluorescent Probe of G-Quadruplex Structures.

G-quadruplex existence was proved in cells by using both antibodies and small molecule fluorescent probes. However, the G-quadruplex probes designed thus far are structure- but not conformation-specific. Recently, a core-extended naphthalene diimide (cex-NDI) was designed and found to provide fluorescent signals of markedly different intensities when bound to G-quadruplexes of different conformations or duplexes. Aiming at evaluating how the fluorescence behaviour of this compound is associated with specific binding modes to the different DNA targets, cex-NDI was here studied in its interaction with hybrid G-quadruplex, parallel G-quadruplex, and B-DNA duplex models by biophysical techniques, molecular docking, and biological assays. cex-NDI showed different binding modes associated with different amounts of stacking interactions with the three DNA targets. The preferential binding sites were the groove, outer quartet, or intercalative site of the hybrid G-quadruplex, parallel G-quadruplex, and B-DNA duplex, respectively. Interestingly, our data show that the fluorescence intensity of DNA-bound cex-NDI correlates with the amount of stacking interactions formed by the ligand with each DNA target, thus providing the rationale behind the conformation-sensitive properties of cex-NDI and supporting its use as a fluorescent probe of G-quadruplex structures. Notably, biological assays proved that cex-NDI mainly localizes in the G-quadruplex-rich nuclei of cancer cells.

Nuclear Factor Related to KappaB Binding Protein (NFRKB) Is a Telomere-Associated Protein and Involved in Liver Cancer Development

Alternative lengthening of telomeres (ALT) is a homologous recombination-based telomere maintenance mechanism activated in 10-15% of human cancers. Although significant progress has been made, the key regulators of the ALT pathway and its role in cancer development remain elusive. Bioinformatics methods were used to predict novel telomere-associated proteins (TAPs) by analysis of large-scale ChIP-Seq data. Immunostaining and fluorescence in situ hybridization experiments were applied to detect the subcellular location of target genes and telomeres. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to examine the expression of targeting genes. Overall survival (OS) analyses were used to evaluate the relationship between gene expression and survival time; immunohistochemistry was used to detect the distribution of target genes in liver cancer tissues. We found that nuclear factor related to kappaB binding protein (NFRKB), a metazoan-specific subunit of the INO80 complex, can associate with telomeres in human ALT cells. Loss of NFRKB induces dysfunction of telomeres and less PML bodies in U2OS cells. In addition, NFRKB is low/moderately expressed in cytoplasm of normal hepatocytes but heavily accumulating in the nucleus of liver cancer cells. Finally, the high expression of NFRKB is associated with short OS time and poor prognosis. NFRKB is a TAP and protects telomeres from DNA damage in ALT cells. It is highly expressed in hepatocellular carcinoma (HCC) cells and predicts a poor prognosis. NFRKB may be a promising prognostic biomarker for the treatment of HCC in the future.

Expression and clinical implications of basic leucine zipper ATF-like transcription factor 2 in breast cancer

BackgroundBasic leucine zipper ATF-like transcription factor 2 (BATF2) has been reported to participate in the occurrence and development of some malignancies. Herein, we aimed to explore the expression pattern and clinical implications of BATF2 in breast cancer (BC).MethodsWe assessed the differences in BATF2 mRNA expression between cancerous and noncancerous tissues in BC using GEPIA and UALCAN data and in BATF2 protein expression pattern using Human Protein Atlas (HPA) data. BATF2 co-expression networks were analyzed in Coexpedia. The association between the differentially expressed BATF2 mRNA and BC prognosis was assessed using UALCAN, OSbrca, and GEPIA databases. In external validations, BATF2 protein expression in BC tissues was quantitated using a tissue microarray and immunohistochemistry (IHC) analysis, and BATF2 mRNA expression in serum and serum-derived exosomes of BC patients using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR).ResultsNo difference in the BATF2 mRNA expression level was found between cancerous and noncancerous tissues in BC based on databases. There were low-to-moderate levels of increases in BATF2 protein expressions in BC cases from the HPA cohort. BATF2 mRNA expression was negatively correlated with androgen receptor (AR) and positively correlated with BRCA2 DNA repair associated (BRCA2), marker of proliferation Ki-67 (Mki67), and tumor protein p53 (TP53) expressions. Generally, BATF2 mRNA exhibited a non-significant association with BC prognosis; yet the subgroup analyses showed that triple-negative breast cancer (TNBC) patients with high BATF2 mRNA expressions had a longer overall survival (OS). Our IHC analysis revealed a positive rate of BATF2 protein expression of 46.90%, mainly located in the nucleus of cancer cells in BC, and the OS of BC patients with high BATF2 protein expressions was prolonged. The positive rates of BATF2 mRNA expressions in the serum and exosomes were 45.00 and 41.67%, respectively. Besides, the AUCs of serum and exosomal BATF2 mRNA for BC diagnosis were 0.8929 and 0.8869, respectively.ConclusionsBC patients exhibit low-to-moderate expressions in BATF2 mRNA expression levels in cancerous tissues. The high BATF2 protein expression can be a potential indicator of a better BC prognosis. Serum and exosomal BATF2 mRNA levels also serve as promising noninvasive biomarkers for BC diagnosis.

Nuclear targeted multimodal 3D-bimetallic Au@Pd nanodendrites promote doxorubicin efficiency in breast cancer therapy

The therapeutic efficiency of doxorubicin (DOX) is dependent on its sufficient accumulation within the nucleus of cancer cells. Taking advantage of the fascinating properties (e.g., multiple dense arrays of hyperbranches and high surface area) of bimetallic nanodendrites as well as the arginine-rich components of protamine sulfate (PS), we herein present a new type of PS-modified [email protected] hydrophobic drug carrier in which DOX can effectively bind to the surface of the [email protected] nanocarrier via non-covalent attachment. Chemical characterization of the synthesized [email protected] showed the successful loading of DOX onto the surface of [email protected] Darkfield and Hyperspectral imaging analysis of the [email protected] demonstrated the time-dependent uptake and prolonged accumulation capability for releasing its DOX cargo inside the nucleus. Specifically, data from the 3D-CytoViva fluorescence imaging did not only track the nanocarrier’s distribution but also confirmed its predominant accumulation in the nuclear compartment through clathrin-mediated endocytosis. Cytotoxicity assay showed that the [email protected] significantly inhibited cancer cell proliferation with maximum DOX release under acidic conditions (pH 4.5 and 5.5) than at normal physiological pH of 7.4. Moreover, the resultant [email protected] DOX nanocarrier efficiently improved the induction of apoptosis in MCF-7 cells compared to free DOX and passive targeted platform. In addition, loss of membrane integrity, mitochondria-apoptotic pathway, and internucleosomal DNA contents from cell cycle progression assay provided insights on the mechanism of cell death. Overall, the [email protected] nanocarrier may be employed for improved subcellular delivery of DOX as well as multimodal visualization in cancer therapy.

Development of Tailor-Made Dendrimer Ternary Complexes for Drug/Gene Co-Delivery in Cancer.

Cancer gene therapy, mediated by non-viral systems, remains a major research focus. To contribute to this field, in this work we reported on the development of dendrimer drug/gene ternary complexes. This innovative approach explored the great capacity of both polyamidoamine (PAMAM)-paclitaxel (PTX) conjugate and polyethylenimine (PEI) polymers to complex a p53-encoding plasmid DNA (pDNA), highlighting the utility of considering two compacting agents. The pDNA complexation capacity has been investigated as function of the nitrogen to phosphate groups ratio (N/P), which revealed to be a tailoring parameter. The physicochemical properties of the conceived ternary complexes were revealed and were found to be promising for cellular transfection. Furthermore, the formulated co-delivery systems demonstrated to be biocompatible. The ternary systems were able of cellular internalization and payload intracellular release. Confocal microscopy studies showed the co-localization of stained pDNA with the nucleus of cancer cells, after transfection mediated by these carriers. From this achievement, p53 gene expression occurred with the production of protein. Moreover, the activation of caspase-3 indicated apoptosis of cancer cells. This work represents a great progress on the design of dendrimer drug/gene co-delivery systems towards a more efficient cancer therapy. In this way, it instigates further in vitro studies concerning the evaluation of their therapeutic potential, expectedly supported by the synergistic effect, in tumoral cells.

Extracellular AGR2 activates neighboring fibroblasts through endocytosis and direct binding to β-catenin that requires AGR2 dimerization and adhesion domains

Anterior gradient 2 (AGR2) is often overexpressed in several types of cancer. AGR2 is cytoplasmic or secreted as an extracellular signal. Intracellular AGR2 properties and role in cancer have been well studied, but its extracellular function is largely unclear. It has been shown that extracellular AGR2 activates endothelial cells and fibroblasts in culture, but the mechanism of AGR2 signaling is not well elucidated. Here, we report that tumor secreted or externally added AGR2 translocates into cytoplasm by endocytosis, binds to β-catenin and further co-translocates to the nucleus in NIH3T3 fibroblasts. Externally added AGR2 also increased β-catenin expression, stability, and accumulation in the nucleus in both fibroblasts and cancer cells. External AGR2 rescued the expression of β-catenin, which was suppressed by EGFR inhibitor AG1478 indicating an alternative pathway to regulate β-catenin independent of EGFR signal. These effects were abolished when a monoclonal antibody against AGR2 was added to the experiments, confirming the effects are caused by AGR2 only. Putting together, our results show that extracellular AGR2 signaling pathway involves endocytosis mediated cellular translocation, direct binding and regulating β-catenin nuclear accumulation. It is also a target against tumor initiated AGR2 signaling to form and maintain tumor microenvironment.

18F-Fluoroestradiol Tumor Uptake Is Influenced by Structural Components in Breast Cancer.

Estrogen receptor (ER) is expressed in the majority of invasive breast cancer and is an important prognostic indicator. The tumor stroma also plays an important role in disease progression. This study evaluated the effect of stromal components on 16α-[18F]-fluoro-17β-estradiol (18F-FES) uptake in breast cancer and proposed a partial-volume correction method for 18F-FES PET based on histopathological analyses. Fifteen patients with biopsy-confirmed breast cancer underwent preoperative 18F-FES PET. Estrogen receptor expression in biopsy specimens was assayed by immunohistochemistry, cellular components in surgical specimens were measured using hematoxylin-eosin staining, and nuclear components in surgical and biopsy specimens were measured using Azan-Mallory staining. The relationship between 18F-FES SUV of the primary tumor and histopathological findings including ER expression, the Allred score, ER-positive cellular component ratio, and ER-positive nuclear component ratio (NCR) was examined. The relationship between stroma-free 18F-FES SUV and ER expression was also examined. 18F-FES uptake was not significantly positively correlated with ER expression (r = 0.44, P = 0.10). 18F-FES uptake was significantly correlated with the Allred score, ER-positive cellular component ratio, and ER-positive NCR in surgical specimens (ρ = 0.60, P = 0.02; r = 0.55, P = 0.03; and r = 0.65, P = 0.01, respectively). 18F-FES uptake was predominantly correlated with ER-positive NCR in biopsy specimens (r = 0.84, P < 0.001). Stroma-free 18F-FES SUV was significantly correlated with ER expression (r = 0.78, P < 0.01). 18F-FES PET predominantly demonstrates the level of ER expression in breast cancer cell nucleus. Although tumor 18F-FES uptake is affected by the degree of stromal components, the partial volume effect on the uptake can be corrected by stroma-volume fraction in Azan-Mallory staining.

H2A.B is a cancer/testis factor involved in the activation of ribosome biogenesis in Hodgkin lymphoma.

Testis-specific regulators of chromatin function are commonly ectopically expressed in human cancers, but their roles are poorly understood. Examination of 81 primary Hodgkin lymphoma (HL) samples showed that the ectopic expression of the eutherian testis-specific histone variant H2A.B is an inherent feature of HL. In experiments using two HL cell lines derived from different subtypes of HL, H2A.B knockdown inhibited cell proliferation. H2A.B was enriched in both nucleoli of these HL cell lines and primary HL samples. We found that H2A.B enhanced ribosomal DNA (rDNA) transcription, was enriched at the rDNA promoter and transcribed regions, and interacted with RNA Pol I. Depletion of H2A.B caused the loss of RNA Pol I from rDNA chromatin. Remarkably, H2A.B was also required for high levels of ribosomal protein gene expression being located at the transcriptional start site and within the gene body. H2A.B knockdown reduced gene body chromatin accessibility of active RNA Pol II genes concurrent with a decrease in transcription. Taken together, our data show that in HL H2A.B has acquired a new function, the ability to increase ribosome biogenesis.

HO-1 and Heme: G-Quadruplex Interaction Choreograph DNA Damage Responses and Cancer Growth.

Many anti-cancer therapeutics lead to the release of danger associated pattern molecules (DAMPs) as the result of killing large numbers of both normal and transformed cells as well as lysis of red blood cells (RBC) (hemolysis). Labile heme originating from hemolysis acts as a DAMP while its breakdown products exert varying immunomodulatory effects. Labile heme is scavenged by hemopexin (Hx) and processed by heme oxygenase-1 (HO-1, Hmox1), resulting in its removal and the generation of biliverdin/bilirubin, carbon monoxide (CO) and iron. We recently demonstrated that labile heme accumulates in cancer cell nuclei in the tumor parenchyma of Hx knockout mice and contributes to the malignant phenotype of prostate cancer (PCa) cells and increased metastases. Additionally, this work identified Hx as a tumor suppressor gene. Direct interaction of heme with DNA G-quadruplexes (G4) leads to altered gene expression in cancer cells that regulate transcription, recombination and replication. Here, we provide new data supporting the nuclear role of HO-1 and heme in modulating DNA damage response, G4 stability and cancer growth. Finally, we discuss an alternative role of labile heme as a nuclear danger signal (NDS) that regulates gene expression and nuclear HO-1 regulated DNA damage responses stimulated by its interaction with G4.