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Cell, Tumor, and Stem Cell Biology

Pancreatic Cancer and Precursor Pancreatic Intraepithelial Neoplasia Lesions Are Devoid of Primary Cilia

E. Scott Seeley, Catherine Carrière, Tobias Goetze, Daniel S. Longnecker and Murray Korc
E. Scott Seeley
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Catherine Carrière
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Tobias Goetze
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Daniel S. Longnecker
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Murray Korc
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DOI: 10.1158/0008-5472.CAN-08-1290 Published January 2009
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    Figure 1.

    Primary cilia are absent in human PDAC and PanIN lesions. A, PanIN-1. B, PanIN-2. C, poorly differentiated PDAC. D, PanIN-2/3. A to D, blue, nuclei. A to C, red, CK19; green, acetylated α-tubulin. D, red, Ki67; green, α-tubulin. Insets: A to D, bottom left, nonneoplastic ducts from adjacent normal region within respective specimens; top right, region of neoplastic ductal epithelium taken at 150% gain and power, revealing a mitotic figure and cytosolic tubules, with absence of ciliary projections. Bar, 20 μm.

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    Figure 2.

    Primary cilia are absent in Kras-dependent murine pancreatic neoplasia. A, left, centroacinar cells from Pdx1-Cre; middle, LSL-KrasG12D (Pdx-Kras) and normal ductule and islet from Pdx1-Cre; right, LSL-KrasG12D;Ink4a/Arflox/lox (Pdx-Kras/Ink4a–deleted) murine pancreata. B, left, mPanIN-1 from Pdx-Kras; middle, mPanIN-2/3 from Pdx-Kras/Ink4A–deleted murine pancreata; right, murine pancreatic cancer from same tissue sample as in middle image. C, centrosomes, stained with an anti-pericentrin antibody, and primary cilia in normal and neoplastic ducts from Pdx1-Cre;LSL-KrasG12D pancreata. Inset, magnified view of cilia and centrosomes from cells immediately below the inset. Staining for CK19, acetylated α-tubulin (AT), insulin, and pericentrin (PC), as indicated. Bar, 20 μm.

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    Figure 3.

    Primary cilia are absent in transitional PanIN lesions. A, intralobular mPanIN from Pdx1-Cre;LSL-KrasG12D pancreas reveals normal-appearing ductal epithelium (solid white lines) and neoplastic ductal epithelium (dashed lines). Yellow arrowheads, transition points between normal and neoplastic epithelia. Is, islets. Red, CK19; green, AT; blue, nuclei. Bar, 50 μm. B, magnified region from A, stained as in A, but with anti-Ki67 antibody in white (white arrowhead). Bar, 50 μm. C, interlobular mPanIN from a Pdx1-Kras pancreas. Staining as in A. Bar, 20 μm. Inset, magnified region from C with addition of Ki67 channel in white (white arrowhead). Bar, 50 μm. D, mPanIN from Nestin-Cre;LSL-KrasG12D pancreas stained for CK19 (red), AT (green), and Ki67 (white arrowheads), revealing the terminal cul-de-sac–like architecture (dashed lines) of a mPanIN lesion appearing to arise from a small ductule (solid lines). Inset, magnified view of ciliated normal ductule from within main image. Bar, 20 μm.

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    Figure 4.

    Preservation and reversal of ciliary assembly defect in Rink-1 pancreatic cancer cells prepared from Pdx1-Cre;LSL-KrasG12D;Ink4a/Arflox/lox tumors. A, RInk-1 cells were cultured for 48 h in the presence of FBS in the absence or presence of the indicated inhibitors (10 μmol/L each). Cells were then fixed at room temperature and stained for microscopy with antibodies to acetylated α-tubulin, detyrosinated α-tubulin (DT), and pericentrin, as indicated. Bar, 10 μm. B, proliferation indices as percentage of cells with nuclear Ki67 and PCNA nuclear positivity following a 48-h incubation with the indicated FBS concentrations. Columns, mean from three independent experiments; bars, SD. C, percent ciliated cells following a 48-h incubation in 0.1% FBS in the absence or presence of the indicated inhibitors (10 μmol/L each). Columns, mean from three independent experiments; bars, SE.

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    Figure 5.

    Primary cilia are present in ADM lesions. A, top, images of H&E-stained regions of normal pancreas and early and late ADM lesions from patients with chronic pancreatitis. Bottom, consecutive serial sections of a late ADM lesion stained with antibodies to CK19, acetylated α-tubulin, and amylase, as indicated. Inset, magnified view of amylase-positive cell from within main image. B, foci of ADM in Nestin-Kras and Pdx-1-Kras pancreata stained with antibodies to CK19, AT, and Ki67, as indicated. C, proliferation in early ADM (left), late ADM (middle), and mPanIN (left) in Pdx1-Kras pancreata stained with antibodies to amylase, AT, and Ki67 as indicated. Bar, 20 μm.

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    Figure 6.

    Quantification of primary cilia and proliferation indices in human and murine PanINs, PDAC, and ADM lesions. Data are the means 9 to 12 fields; bars, SD. N, total number of cells scored per grouping.

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Cancer Research: 69 (2)
January 2009
Volume 69, Issue 2
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Pancreatic Cancer and Precursor Pancreatic Intraepithelial Neoplasia Lesions Are Devoid of Primary Cilia
E. Scott Seeley, Catherine Carrière, Tobias Goetze, Daniel S. Longnecker and Murray Korc
Cancer Res January 15 2009 (69) (2) 422-430; DOI: 10.1158/0008-5472.CAN-08-1290

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Pancreatic Cancer and Precursor Pancreatic Intraepithelial Neoplasia Lesions Are Devoid of Primary Cilia
E. Scott Seeley, Catherine Carrière, Tobias Goetze, Daniel S. Longnecker and Murray Korc
Cancer Res January 15 2009 (69) (2) 422-430; DOI: 10.1158/0008-5472.CAN-08-1290
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