α2 Integrin-Dependent Suppression of Pancreatic Adenocarcinoma Cell Invasion Involves Ectodomain Regulation of Kallikrein-Related Peptidase-5

Previous reports demonstrate that the α2-integrin (α2) mediates pancreatic ductal adenocarcinoma (PDAC) cell interactions with collagens. We found that while well-differentiated cells use α2 exclusively to adhere and migrate on collagenI, poorly differentiated PDAC cells demonstrate reduced reliance on, or complete loss of, α2. Since well-differentiated PDAC lines exhibit reducedin vitroinvasion and α2-blockade suppressed invasion of well-differentiated lines exclusively, we hypothesized that α2 may suppress the malignant phenotype in PDAC. Accordingly, ectopic expression of α2 retardedin vitroinvasion and maintenance on collagenI exacerbated this effect. Affymetrix profiling revealed that kallikrein-related peptidase-5 (KLK5) was specifically upregulated by α2, and reduced α2 and KLK5 expression was observed in poorly differentiated PDAC cellsin situ. Accordingly, well-differentiated PDAC lines express KLK5, and KLK5 blockade increased the invasion of KLK5-positive lines. The α2-cytoplasmic domain was dispensable for these effects, demonstrating that the α2-ectodomain and KLK5 coordinately regulate a less invasive phenotype in PDAC.

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Inside-Out Regulation of L1 Conformation, Integrin Binding, Proteolysis, and Concomitant Cell Migration

Molecular Biology of the Cell ◽

10.1091/mbc.e09-10-0900 ◽

2010 ◽

Vol 21 (10) ◽

pp. 1671-1685 ◽

Cited By ~ 10

Author(s):

Maxine M. Chen ◽

Chia-Yao Lee ◽

Hyuma A. Leland ◽

Grace Y. Lin ◽

Anthony M. Montgomery ◽

...

Keyword(s):

Cell Migration ◽

Ductal Adenocarcinoma ◽

Cell Phenotype ◽

Poorly Differentiated ◽

Differential Binding ◽

A Disintegrin And Metalloproteinase ◽

Cell Adhesion Molecule L1 ◽

Inside Out

Previous reports on the expression of the cell adhesion molecule L1 in pancreatic ductal adenocarcinoma (PDAC) cells range from absent to high. Our data demonstrate that L1 is expressed in poorly differentiated PDAC cells in situ and that threonine-1172 (T1172) in the L1 cytoplasmic domain exhibits steady-state saturated phosphorylation in PDAC cells in vitro and in situ. In vitro studies support roles for casein kinase II and PKC in this modification, consistent with our prior studies using recombinant proteins. Importantly, T1172 phosphorylation drives, or is associated with, a change in the extracellular structure of L1, consistent with a potential role in regulating the shift between the closed conformation and the open, multimerized conformation of L1. We further demonstrate that these distinct conformations exhibit differential binding to integrins αvβ3 and αvβ5 and that T1172 regulates cell migration in a matrix-specific manner and is required for a disintegrin and metalloproteinase-mediated shedding of the L1 ectodomain that has been shown to regulate cell migration. These data define a specific role for T1172 of L1 in regulating aspects of pancreatic adenocarcinoma cell phenotype and suggest the need for further studies to elucidate the specific ramifications of L1 expression and T1172 phosphorylation in the pathobiology of pancreatic cancer.

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Resistance of differentiated human airway epithelium to infection by rhinovirus

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00300.2003 ◽

2004 ◽

Vol 286 (2) ◽

pp. L373-L381 ◽

Cited By ~ 75

Author(s):

N. Lopez-Souza ◽

G. Dolganov ◽

R. Dubin ◽

L. A. Sachs ◽

L. Sassina ◽

...

Keyword(s):

Viral Infection ◽

Airway Epithelium ◽

Primary Cultures ◽

Viral Rna ◽

Human Airway ◽

Human Airway Epithelium ◽

Differentiated Cells ◽

Poorly Differentiated ◽

Well Differentiated

Virtually all in vitro studies of the effects of rhinovirus on human airway epithelium have used cells grown under conditions known to produce low levels of differentiation. The relevance of the results to native epithelium is questionable. Here we grew primary cultures of human tracheal or nasal epithelium under three conditions. One condition produced pseudostratified, mucociliary cells virtually indistinguishable from native epithelium. The other two conditions produced undifferentiated squamous cells lacking cilia. Cells were infected for 6 h with rhinovirus-16. After a 24-h incubation period, we determined levels of viral RNA in the cells, numbers of infectious viral particles released in the mucosal medium, expression of a variety of epithelial cytokines and other proteins, release of IL-6 and IL-8, and transepithelial electrical resistance and voltage. After infection, levels of viral RNA in the poorly differentiated cells were 30 or 130 times those in the differentiated. Furthermore, expression of mRNA for inflammatory cytokines, release of infectious particles, and release of IL-6 and IL-8 were closely correlated with the degree of viral infection. Thus well-differentiated cells are much more resistant to viral infection and its functional consequences than are poorly differentiated cells from the same source.

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Motility of leukemic lymphocytes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100159382 ◽

1990 ◽

Vol 48 (3) ◽

pp. 368-369

Author(s):

Manoj Raje ◽

Karvita B. Ahluwalia

Keyword(s):

Quantitative Data ◽

Laser Doppler Velocimetry ◽

Acute Lymphocytic Leukemia ◽

Lymphocytic Leukemia ◽

Cellular Space ◽

Differentiated Cells ◽

Poorly Differentiated ◽

Solid Tissues ◽

Well Differentiated ◽

Reduced Mobility

In Acute Lymphocytic Leukemia motility of lymphocytes is associated with dissemination of malignancy and establishment of metastatic foci. Normal and leukemic lymphocytes in circulation reach solid tissues where due to in adequate perfusion some cells get trapped among tissue spaces. Although normal lymphocytes reenter into circulation leukemic lymphocytes are thought to remain entrapped owing to reduced mobility and form secondary metastasis. Cell surface, transmembrane interactions, cytoskeleton and level of cell differentiation are implicated in lymphocyte mobility. An attempt has been made to correlate ultrastructural information with quantitative data obtained by Laser Doppler Velocimetry (LDV). TEM of normal & leukemic lymphocytes revealed heterogeneity in cell populations ranging from well differentiated (Fig. 1) to poorly differentiated cells (Fig. 2). Unlike other cells, surface extensions in differentiated lymphocytes appear to originate by extrusion of large vesicles in to extra cellular space (Fig. 3). This results in persistent unevenness on lymphocyte surface which occurs due to a phenomenon different from that producing surface extensions in other cells.

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Factors affecting overall survival in ductal adenocarcinoma of the pancreatic head. Experience of one center

Malignant tumours ◽

10.18027/2224-5057-2021-11-1-20-28 ◽

2021 ◽

Vol 11 (1) ◽

pp. 20-28

Author(s):

D. M. Kuchin ◽

Ya. I. Kolesnik ◽

H. G. Torgomyan ◽

V. E. Zagainov

Keyword(s):

Overall Survival ◽

Adjuvant Chemotherapy ◽

Vascular Invasion ◽

Survival Rates ◽

Pancreatic Head ◽

Ductal Adenocarcinoma ◽

Factors Affecting ◽

Poorly Differentiated ◽

Well Differentiated ◽

Major Factors

Purpose. To identify major factors affecting the overall survival (OS). To select the cohort of patients with the best prognosis.Materials and methods. A retrospective analysis included data of 268 patients, 128 men and 140 women, with median age of 59±10,53 (30 to 83) years. For multivariate analysis of survival, patients were selected who underwent pancreaticoduodenectomy (PD) for ductal adenocarcinoma of the pancreatic head.Results. Our study demonstrated that histologically verified vascular invasion (detected only in 30 % of patients who underwent PD with resection of the major vessels) statistically significantly affected the OS. The increased CA19-9 level over 500 U / L (detected in 32,3 % of cases) is the factor that significantly worsens the OS. Patients with high grade adenocarcinoma have significantly better survival rates compared with patients who have moderately or poorly differentiated adenocarcinoma (p = 0.014; median 26 months, 95 % CI 4.4–47.6 versus median 17 months, 95 % CI 15–19, an median: 13 months, 95 % CI 5–21, respectively). Also, the use of adjuvant chemotherapy has a positive effect on long-term outcomes (p = 0.0001; median 26 months, 95 % CI 21.7–30.3 versus median 13 months, 95 % CI 11.3–14.7).Conclusion. A well-differentiated tumor and the use of adjuvant chemotherapy significantly increase the OS of patients. Poorly differentiated tumor, CA19-9 level over 500 U / mL and the histologically confirmed vascular invasion significantly worsen the prognosis of these patients.

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FUS-induced circRHOBTB3 facilitates cell proliferation via miR-600/NACC1 mediated autophagy response in pancreatic ductal adenocarcinoma

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02063-w ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Taoyue Yang ◽

Peng Shen ◽

Qun Chen ◽

Pengfei Wu ◽

Hao Yuan ◽

...

Keyword(s):

Cell Proliferation ◽

Pancreatic Ductal Adenocarcinoma ◽

Cell Lines ◽

Rna Binding ◽

Ductal Adenocarcinoma ◽

Luciferase Reporter ◽

Circular Rnas ◽

Pdac Cell

Abstract Background Circular RNAs (circRNAs) are becoming a unique member of non-coding RNAs (ncRNAs) with emerging evidence of their regulatory roles in various cancers. However, with regards to pancreatic ductal adenocarcinoma (PDAC), circRNAs biological functions remain largely unknown and worth investigation for potential therapeutic innovation. Methods In our previous study, next-generation sequencing was used to identify differentially expressed circRNAs in 3 pairs of PDAC and adjacent normal tissues. Further validation of circRHOBTB3 expression in PDAC tissues and cell lines and gain-and-loss function experiments verified the oncogenic role of circRHOBTB3. The mechanism of circRHOBTB3 regulatory role was validated by pull-down assays, RIP, luciferase reporter assays. The autophagy response of PANC-1 and MiaPaca-2 cells were detected by mCherry-GFP-LC3B labeling and confocal microscopy, transmission electron microscopy and protein levels of LC3B or p62 via Western blot. Results circRHOBTB3 is highly expressed in PDAC cell lines and tissues, which also promotes PDAC autophagy and then progression in vitro and in vivo. Mechanistically, circRHOBTB3 directly binds to miR-600 and subsequently acts as a miRNA-sponge to maintain the expression level of miR-600-targeted gene NACC1, which facilitates the autophagy response of PDAC cells for adaptation of proliferation via Akt/mTOR pathway. Moreover, the RNA-binding protein FUS (FUS) directly binds to pre-RHOBTB3 mRNA to mediate the biogenesis of circRHOBTB3. Clinically, circRHOBTB3, miR-600 and NACC1 expression levels are correlated with the prognosis of PDAC patients and serve as independent risk factors for PDAC patients. Conclusions FUS-mediated circRHOBTB3 functions as a tumor activator to promote PDAC cell proliferation by modulating miR-600/NACC1/Akt/mTOR axis regulated autophagy.

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DPre: computational identification of differentiation bias and genes underlying cell type conversions

Bioinformatics ◽

10.1093/bioinformatics/btz789 ◽

2019 ◽

Author(s):

Simon Steffens ◽

Xiuling Fu ◽

Fangfang He ◽

Yuhao Li ◽

Isaac A Babarinde ◽

...

Keyword(s):

Ectopic Expression ◽

Large Cell ◽

Cell Types ◽

Supplementary Information ◽

Computational Techniques ◽

Cell Type ◽

Mixed Cell ◽

Directional Bias ◽

Differentiated Cells

Abstract Summary Cells are generally resistant to cell type conversions, but can be converted by the application of growth factors, chemical inhibitors and ectopic expression of genes. However, it remains difficult to accurately identify the destination cell type or differentiation bias when these techniques are used to alter cell type. Consequently, there is demand for computational techniques that can help researchers understand both the cell type and differentiation bias. While advanced tools identifying cell types exist for single cell data and the deconvolution of mixed cell populations, the problem of exploring partially differentiated cells of indeterminate transcriptional identity has not been addressed. To fill this gap, we developed driver-predictor, which relies on scoring per gene transcriptional similarity between RNA-Seq datasets to reveal directional bias of differentiation. By comparing against large cell type transcriptome libraries or a desired target expression profile, the tool enables the user to visualize both the changes in transcriptional identity as well as the genes accounting for the cell type changes. This software will be a powerful tool for researchers to explore in vitro experiments that involve cell type conversions. Availability and implementation Source code is open source under the MIT license and is freely available on https://github.com/LoaloaF/DPre. Supplementary information Supplementary data are available at Bioinformatics online.

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Glutathione S-Transferase pi-1 Knockdown Reduces Pancreatic Ductal Adenocarcinoma Growth by Activating Oxidative Stress Response Pathways

Cancers ◽

10.3390/cancers12061501 ◽

2020 ◽

Vol 12 (6) ◽

pp. 1501

Author(s):

Rahul R. Singh ◽

Jiyan Mohammad ◽

Megan Orr ◽

Katie M. Reindl

Keyword(s):

Oxidative Stress ◽

Cell Growth ◽

Pancreatic Ductal Adenocarcinoma ◽

Cell Cycle Progression ◽

Map Kinases ◽

Ductal Adenocarcinoma ◽

Glutathione S Transferase ◽

Pdac Cell ◽

Ki 67

Glutathione S-transferase pi-1 (GSTP1) plays an important role in regulating oxidative stress by conjugating glutathione to electrophiles. GSTP1 is overexpressed in breast, colon, lung, and prostate tumors, where it contributes to tumor progression and drug resistance; however, the role of GSTP1 in pancreatic ductal adenocarcinoma (PDAC) is not well understood. Using shRNA, we knocked down GSTP1 expression in three different PDAC cell lines and determined the effect on cell proliferation, cell cycle progression, and reactive oxygen species (ROS) levels. Our results show GSTP1 knockdown reduces PDAC cell growth, prolongs the G0/G1 phase, and elevates ROS in PDAC cells. Furthermore, GSTP1 knockdown results in the increased phosphorylation of c-Jun N-terminal kinase (JNK) and c-Jun and the decreased phosphorylation of extracellular signal-regulated kinase (ERK), p65, the reduced expression of specificity protein 1 (Sp1), and the increased expression of apoptosis-promoting genes. The addition of the antioxidant glutathione restored cell viability and returned protein expression levels to those found in control cells. Collectively, these data support the working hypothesis that the loss of GSTP1 elevates oxidative stress, which alters mitogen-activated protein (MAP) kinases and NF-κB signaling, and induces apoptosis. In support of these in vitro data, nude mice bearing orthotopically implanted GSTP1-knockdown PDAC cells showed an impressive reduction in the size and weight of tumors compared to the controls. Additionally, we observed reduced levels of Ki-67 and increased expression of cleaved caspase-3 in GSTP1-knockdown tumors, suggesting GSTP1 knockdown impedes proliferation and upregulates apoptosis in PDAC cells. Together, these results indicate that GSTP1 plays a significant role in PDAC cell growth and provides support for the pursuit of GSTP1 inhibitors as therapeutic agents for PDAC.

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Radial endobronchial ultrasound images for ground-glass opacity pulmonary lesions

European Respiratory Journal ◽

10.1183/09031936.00167914 ◽

2015 ◽

Vol 45 (6) ◽

pp. 1661-1668 ◽

Cited By ~ 26

Author(s):

Takehiro Izumo ◽

Shinji Sasada ◽

Christine Chavez ◽

Yuji Matsumoto ◽

Takaaki Tsuchida

Keyword(s):

Ground Glass Opacity ◽

Endobronchial Ultrasound ◽

Ground Glass ◽

Ultrasound Images ◽

Poorly Differentiated Adenocarcinoma ◽

Pulmonary Lesions ◽

Poorly Differentiated ◽

Peripheral Pulmonary Lesions ◽

Well Differentiated

Radial endobronchial ultrasound (R-EBUS) is a useful tool for precise localisation of peripheral pulmonary lesions, but there have been no detailed reports about the use of R-EBUS images for ground-glass opacity (GGO).The R-EBUS images of 116 patients with GGO, who were diagnosed as having adenocarcinoma by R-EBUS with a guide sheath (EBUS-GS), were compared with the respective chest computed tomography findings. In 103 patients, R-EBUS images were correlated with the histological surgical specimens.R-EBUS images of GGO were identified based on the internal structure of the lesion and classified into two groups. Blizzard showed an enlarged, diffuse hyperintense acoustic shadow. Mixed blizzard showed a combination of blizzard and some diffuse heterogeneity with several hyperechoic dots and vessels. All pure GGO lesions (nine out of nine) were blizzard on R-EBUS. For part-solid GGOs, the percentage of mixed blizzard was inversely related to the amount of the GGO component. Histological findings from surgery revealed that all blizzard lesions were on the spectrum of adenocarcinoma in situ to well differentiated adenocarcinoma while majority (33 out of 64) of mixed blizzard lesions were moderately to poorly differentiated adenocarcinoma.R-EBUS types are important to locate GGOs prior to transbronchial sampling with EBUS-GS.

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Dynamics of morphological changes in neural cell culture with a model of neurotrauma under the influence of conditioned media of the rat fetal brain neurogenic cells

Cell and Organ Transplantology ◽

10.22494/cot.v8i2.114 ◽

2020 ◽

Vol 8 (2) ◽

Author(s):

E. Pedachenko ◽

◽

L. Liubich ◽

L. Staino ◽

D. Egorova ◽

...

Keyword(s):

Cell Culture ◽

Mitotic Activity ◽

Fetal Brain ◽

Conditioned Media ◽

Differentiated Cells ◽

Cell Processes ◽

Poorly Differentiated ◽

Single Addition ◽

Vitro Model

A potential strategy for recovery and regeneration of brain damage due to traumatic brain injury is considered to be the transplantation of neurogenic stem and/or progenitor cells (NSCs/NPCs). The key factors of the regenerative non-targeted effects of NSCs/NPCs (so-called bystander effects) include the signal molecules produced by them into the extracellular environment (secretome). The purpose is to study the regenerative bystander effects of rat fetal brain neurogenic cells (FBNCs) in the in vitro model of neurotrauma. Materials and methods. In cell culture of FBNCs from rat fetuses (E14-16), neurotrauma was modeled in vitro by mechanical scratching of monolayer and conditioned medium obtained from 24-h cultures of rat FBNCs was added. Cell phenotype was evaluated by morphological features and by immunocytochemical staining for Nestin and GFAP. The density and length of processes, migration capacity, the cell growth rate and monolayer density in the scratched area were compared. Morphometric study included analysis of the width of the scratched area, the number of migrating cells, the distance of migration and mitotic activity in the intact monolayer. Results. Under the conditions of the nutrient medium of standard composition in the scratched area the signs of endogenous regeneration are shown during 24-48 h of cultivation. The overgrowth of cell processes from monolayer and short distance migration of single undifferentiated or poorly differentiated cells were shown. In the next 72-96 h of observation, the degeneration of migrated cells and processes in the scratched area was detected. Under the influence of conditioned media from 24-h cultures of FBNCs by single addition immediately after scratching at dose of 0.1 mg/ml for protein content the stimulation of regeneration were detected up to 96 hours of cultivation. The migration of cell processes from the monolayer simultaneously with undifferentiated or poorly differentiated cells at 24 hours was shown. The formation of cell clusters and their differentiation (at 48 h), as well as migration of differentiated cells with partial or complete overgrowth of scratched area (72-96 h) were observed. The morphological signs of degeneration of migrated cells in the scratched area appeared only on the 8th day of cultivation. Conditioned media does not affect qualitative and quantitative properties of the culture of rat FBNCs in the intact area where mitotic activity was average. Conclusions. Conditioned medium from 24-h cultures of rat FBNC can stimulate reparation in the in vitro model of neurotrauma in neural cell culture for at least 7 days at a single addition, without affecting the cellular composition and mitotic activity of the intact monolayer.

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Differentiation potentiality of rat visceral yolk sac in organ culture

Development ◽

10.1242/dev.80.1.127 ◽

1984 ◽

Vol 80 (1) ◽

pp. 127-136

Author(s):

Y. L. Lu ◽

H. Sobis ◽

L. Van Hove ◽

M. Vandeputte

Keyword(s):

Organ Culture ◽

In Vitro Culture ◽

Yolk Sac ◽

Trophoblast Cells ◽

Differentiated Cells ◽

Visceral Yolk Sac ◽

Poorly Differentiated

Visceral yolk sacs removed at day 12 of pregnancy in the rat were kept in organ culture for as long as 28 days. During this in vitro culture, proliferation of the endoderm and the mesoderm as well as of poorly differentiated cells was observed. The latter displayed neither the characteristics of endodermal nor mesodermal cells and their presence was frequently associated with the development of giant trophoblast cells. The hypothesis is proposed that these trophoblast cells originate from these poorly differentiated cells that acquire in vivo and in vitro the potentiality to differentiate.

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