Further delineation of the continuous human neoplastic enterochromaffin cell line, KRJ-I, and the inhibitory effects of lanreotide and rapamycin

Kidd, Mark; Eick, Geeta; Modlin, Irvin M.; Pfragner, Roswitha; Champaneria, Manish C.; Murren, John R.

doi:10.1677/jme.1.02037

Cited by 44 publications

(86 citation statements)

References 61 publications

(32 reference statements)

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“…Chromogranin A however was not measurable in the conditioned media from KRJ-I cells (data not shown) even after 8-h exposure to agonists; the mRNA and protein were detectable by RT-PCR (unpublished observation-J. Ham) and immunofluorescence, respectively [18]. …”

Section: Resultsmentioning

confidence: 92%

Adenosine A2A and A2B receptor expression in neuroendocrine tumours: potential targets for therapy

Kalhan

Gharibi

Vazquez

et al. 2011

Purinergic Signalling

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The clinical management of neuroendocrine tumours is complex. Such tumours are highly vascular suggesting tumour-related angiogenesis. Adenosine, released during cellular stress, damage and hypoxia, is a major regulator of angiogenesis. Herein, we describe the expression and function of adenosine receptors (A 1 , A 2A , A 2B and A 3 ) in neuroendocrine tumours. Expression of adenosine receptors was investigated in archival human neuroendocrine tumour sections and in two human tumour cell lines, BON-1 (pancreatic) and KRJ-I (intestinal). Their function, with respect to growth and chromogranin A secretion was carried out in vitro. Immunocytochemical data showed that A 2A and A 2B receptors were strongly expressed in 15/15 and 13/18 archival tumour sections. Staining for A 1 (4/18) and A 3 (6/18) receptors was either very weak or absent. In vitro data showed that adenosine stimulated a three-to fourfold increase in cAMP levels in BON-1 and KRJ-1 cells. The non-selective adenosine receptor agonist (adenosine-5′N-ethylcarboxamide, NECA) and the A 2A R agonist (CGS21680) stimulated cell proliferation by up to 20-40% which was attenuated by A 2B (PSB603 and MRS1754) and A 2A (SCH442416) receptor selective antagonists but not by the A 1 receptor antagonist (PSB36). Adenosine and NECA stimulated a twofold increase in chromogranin A secretion in BON-1 cells. Our data suggest that neuroendocrine tumours predominantly express A 2A and A 2B adenosine receptors; their activation leads to increased proliferation and secretion of chromogranin A. Targeting adenosine signal pathways, specifically inhibition of A 2 receptors, may thus be a useful addition to the therapeutic management of neuroendocrine tumours.

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Section: Resultsmentioning

confidence: 92%

Adenosine A2A and A2B receptor expression in neuroendocrine tumours: potential targets for therapy

Kalhan

Gharibi

Vazquez

et al. 2011

Purinergic Signalling

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“…31,36 Cells were incubated at 378C with 5% carbon dioxide. Population doubling level (PDL) 37 25 to 30 was used for all experiments.…”

Section: Neoplastic Ec Cells (Krj-i Cell Line)mentioning

confidence: 99%

“…31,36 TGFb1 (Santa Cruz Biotechnology, Santa Cruz, Calif [hBA-112, sc-4561]) was added at concentrations ranging from 1 pM to 100 pM (n ¼ 8 wells for each concentration) 38 and cells were incubated for 72 hours. MTT was added (final concentration 0.5 mg/mL per well) and incubated for 3 hours (378C at 5% carbon dioxide), and the reaction stopped by adding acid-isopropanol, and the formazan dye solubilized.…”

Section: Proliferation Studiesmentioning

confidence: 99%

“…Optical density was read at 595 nanometers (nm) using a microplate reader (BioRad, Hercules, Calif; 3500). 36 Real-Time Polymerase Chain Reaction RNA was extracted from 1 3 10 6 acutely isolated normal EC cells (n ¼ 3) or 5 3 10 6 KRJ-I cells in log phase growth (n ¼ 3) (TRIZOL, Invitrogen, La Jolla, Calif) and cleaned (Qiagen RNeasy kit and DNeasy Tissue kit, Qiagen, Valencia, Calif ) to minimize contaminating genomic DNA. RNA (2 lg) was converted to cDNA (High Capacity cDNA Archive Kit; Applied Biosystems [ABI], Foster City, Calif).…”

Section: Proliferation Studiesmentioning

confidence: 99%

“…Normal EC cells or KRJ-I cells (5 3 10 4 cells) were fixed in methanol and pipetted onto frosted microscope slides stained with a series of antibodies including anti-TGFbRII (1:1000, sc-220), neuroendocrine-specific antibodies (mouse chromogranin A 31,36 After washing (0.1% Tween/PBS), cells were stained with secondary antibody (FITC-antimouse/rabbit, 1:100; Promega, Madison, Wis) (1 hour at RT). Fluorescence microscopy was used to visualize and count the number of positive cells, with 1 3 10 3 cells counted for each antibody.…”

Section: Immunostainingmentioning

confidence: 99%

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Small bowel carcinoid (enterochromaffin cell) neoplasia exhibits transforming growth factor–β1‐mediated regulatory abnormalities including up‐regulation of C‐Myc and MTA1

Kidd

Modlin

Pfragner

et al. 2007

Cancer

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Objective To examine whether and to what extent the intracellular trafficking features of HLA–B*2705, which is associated with the development of spondylarthritis (SpA), differ from those of HLA–B*2709 and HLA–B*0702, which are not associated with SpA. Methods HeLa cells were transfected with complementary DNA encoding for HLA–B proteins fused to Renilla luciferase or yellow fluorescent protein. The subcellular distribution of properly folded and unfolded/misfolded HLA–B proteins was examined by flow cytometry and confocal microscopy of cells labeled with ME1 and HC‐10 antibodies, respectively. HLA–B/HLA–B interactions were monitored in endoplasmic reticulum (ER)– and plasma membrane–enriched subcellular fractions, by bioluminescence resonance energy transfer (BRET). Results All 3 HLA–B alleles displayed a similar distribution pattern (properly folded heavy chain at the cell surface, unfolded/misfolded proteins only in the cytoplasm). By means of BRET, we provided evidence that both HLA–B*2705 and HLA–B*2709 formed more oligomers in the ER and the plasma membrane than did HLA–B*0702. The propensity of HLA–B*2705 to form oligomers in the ER was partly attributable to residue Cys67 of the molecule. For all 3 alleles, increased expression of HLA–B proteins was associated with intracytoplasmic accumulation of unfolded/misfolded proteins and intracellular vesicles, probably corresponding to expanded ER–Golgi intermediate compartments, in which these proteins accumulated together with the stress sensor BiP. Conclusion Our results suggest that the difference in disease susceptibility conferred by HLA–B*2705 and HLA–B*2709 cannot be explained by their different propensity to form dimers or misfolded proteins, thus presumably implicating other, still unknown factors.

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Inhibition of proliferation of small intestinal and bronchopulmonary neuroendocrine cell lines by using peptide analogs targeting receptors

Kidd

Schally

Pfragner

et al. 2008

Cancer

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BACKGROUND. Currently, no consistently effective therapy is available to inhibit cell proliferation or metastasis of neuroendocrine tumor (NET) disease. The effects of 4 novel peptides were analyzed: a targeted cytotoxic analog of luteinizing hormone‐releasing hormone (LH‐RH) analog (AN‐152), a targeted cytotoxic analog of somatostatin (AN‐238), and 2 antagonists of growth hormone‐releasing hormone (GH‐RH) on 3 NET (carcinoid) cell lines that expressed respective peptide receptors. METHODS. The effects of the compounds were evaluated on cell proliferation in vitro using MTT uptake and Ki67 expression, apoptosis (caspase 3 expression and activity), and cell cycle parameters (DNA distribution). RESULTS. Proliferation of the LH‐RH receptor‐expressing lung NET, NCI‐H720 line, was inhibited 2‐fold by AN‐152 containing doxorubicin compared with the chemotherapy alone (IC50 of 9.1 nM vs 24 nM). This was associated with a reduction in Ki67 transcript and an increase in both caspase 3 mRNA levels and activity. Proliferation of the GH‐RH receptor expressing lung NET, NCI‐H727 line, was inhibited by both GH‐RH antagonists, the effects being mediated through changes in Ki67 expression, but not in caspase 3‐mediated apoptosis. The small intestinal NET, KRJ‐I line, was 8× more sensitive to inhibition by AN‐238 than to 2‐pyrolino‐doxorubicin, reflected by increased caspase 3 transcript as well as activity. AN‐238‐mediated growth inhibition culminated in complete G1 arrest. CONCLUSIONS. The data demonstrate GH‐RH antagonists or peptide‐linked antineoplastic agents such as AN‐152 and AN‐238 are effective inhibitors of NET proliferation in vitro. Because peptide receptors such as those for GH‐RH, LH‐RH, and SST subtypes are commonly expressed by NETs, the development of antineoplastic agents targeted to specific tumor receptors may provide a more efficacious strategy than systemic chemotherapeutic agents currently in use. Cancer 2008. © 2008 American Cancer Society.

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Further delineation of the continuous human neoplastic enterochromaffin cell line, KRJ-I, and the inhibitory effects of lanreotide and rapamycin

Cited by 44 publications

References 61 publications

Adenosine A2A and A2B receptor expression in neuroendocrine tumours: potential targets for therapy

Adenosine A2A and A2B receptor expression in neuroendocrine tumours: potential targets for therapy

Small bowel carcinoid (enterochromaffin cell) neoplasia exhibits transforming growth factor–β1‐mediated regulatory abnormalities including up‐regulation of C‐Myc and MTA1

Inhibition of proliferation of small intestinal and bronchopulmonary neuroendocrine cell lines by using peptide analogs targeting receptors

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