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Amyloid precursor-like protein 2 (APLP2) affects the actin cytoskeleton and increases pancreatic cancer growth and metastasis.

Pandey P, Rachagani S, Das S, Seshacharyulu P, Sheinin Y, Naslavsky N, Pan Z, Smith BL, Peters HL, Radhakrishnan P, McKenna NR, Giridharan SS, Haridas D, Kaur S, Hollingsworth MA, MacDonald RG, Meza JL, Caplan S, Batra SK, Solheim JC - Oncotarget (2015)

Bottom Line: Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine.Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade.Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA.

ABSTRACT
Amyloid precursor-like protein 2 (APLP2) is aberrantly expressed in pancreatic cancer. Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine. Examination of matched human primary tumor-liver metastasis pairs showed that 38.1% of the patients had positive APLP2 expression in both the primary tumor and the corresponding liver metastasis. Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade. Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells. Down-regulation of APLP2 decreased the weight and metastasis of orthotopically transplanted pancreatic tumors in nude mice.

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APLP2 down-regulation alters the morphology of the actin cytoskeletonS2-013-APLP2-shRNA cells (without or with APLP2 down-regulation by Dox) were grown on glass coverslips, fixed, permeabilized, and stained with rhodamine phalloidin (for F-actin filaments) (Thermo Fisher Scientific), and visualized by confocal microscopy. The images were taken with a Zeiss LSM 5 Pascal confocal microscope (Thornwood, NY, USA), using a ×63 1.4 numerical aperture lens and appropriate filters.
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Figure 4: APLP2 down-regulation alters the morphology of the actin cytoskeletonS2-013-APLP2-shRNA cells (without or with APLP2 down-regulation by Dox) were grown on glass coverslips, fixed, permeabilized, and stained with rhodamine phalloidin (for F-actin filaments) (Thermo Fisher Scientific), and visualized by confocal microscopy. The images were taken with a Zeiss LSM 5 Pascal confocal microscope (Thornwood, NY, USA), using a ×63 1.4 numerical aperture lens and appropriate filters.

Mentions: Actin structures are vital to the formation of invadopodia and lamellopodia, which permit tumor cell motility and metastasis [11-14]. In previous studies, APLP2 was found to associate with Fe65 adaptors that interact with the cytoskeleton through the mammalian homolog of the Drosophila Enabled gene (Mena or Enah) or the enabled/vasodilator-stimulated phosphoprotein-like protein (Evl) [15-16]. Both Mena and Evl direct actin arrangement and promote extension of actin filaments [17]. Since APLP2 influences pancreatic cancer cell invasion and migration (Figures 3B,C, Supplementary Figures 1, 2A, and 2B), we investigated whether down-regulation of APLP2 expression influenced the structure of the actin cytoskeleton in pancreatic cancer cells. We observed that Dox-treated S2-013-APLP2-shRNA cells have altered cytoskeletal morphology with substantial rearrangement of the actin cytoskeleton, as shown by staining with rhodamine phalloidin for visualization of actin filaments (Figure 4). Indeed, the APLP2-positive cells tend to display high levels of cortical actin, whereas the cells with APLP2 expression knocked down have less cortical actin and more intracellular microfilaments (Figure 4).


Amyloid precursor-like protein 2 (APLP2) affects the actin cytoskeleton and increases pancreatic cancer growth and metastasis.

Pandey P, Rachagani S, Das S, Seshacharyulu P, Sheinin Y, Naslavsky N, Pan Z, Smith BL, Peters HL, Radhakrishnan P, McKenna NR, Giridharan SS, Haridas D, Kaur S, Hollingsworth MA, MacDonald RG, Meza JL, Caplan S, Batra SK, Solheim JC - Oncotarget (2015)

APLP2 down-regulation alters the morphology of the actin cytoskeletonS2-013-APLP2-shRNA cells (without or with APLP2 down-regulation by Dox) were grown on glass coverslips, fixed, permeabilized, and stained with rhodamine phalloidin (for F-actin filaments) (Thermo Fisher Scientific), and visualized by confocal microscopy. The images were taken with a Zeiss LSM 5 Pascal confocal microscope (Thornwood, NY, USA), using a ×63 1.4 numerical aperture lens and appropriate filters.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4385836&req=5

Figure 4: APLP2 down-regulation alters the morphology of the actin cytoskeletonS2-013-APLP2-shRNA cells (without or with APLP2 down-regulation by Dox) were grown on glass coverslips, fixed, permeabilized, and stained with rhodamine phalloidin (for F-actin filaments) (Thermo Fisher Scientific), and visualized by confocal microscopy. The images were taken with a Zeiss LSM 5 Pascal confocal microscope (Thornwood, NY, USA), using a ×63 1.4 numerical aperture lens and appropriate filters.
Mentions: Actin structures are vital to the formation of invadopodia and lamellopodia, which permit tumor cell motility and metastasis [11-14]. In previous studies, APLP2 was found to associate with Fe65 adaptors that interact with the cytoskeleton through the mammalian homolog of the Drosophila Enabled gene (Mena or Enah) or the enabled/vasodilator-stimulated phosphoprotein-like protein (Evl) [15-16]. Both Mena and Evl direct actin arrangement and promote extension of actin filaments [17]. Since APLP2 influences pancreatic cancer cell invasion and migration (Figures 3B,C, Supplementary Figures 1, 2A, and 2B), we investigated whether down-regulation of APLP2 expression influenced the structure of the actin cytoskeleton in pancreatic cancer cells. We observed that Dox-treated S2-013-APLP2-shRNA cells have altered cytoskeletal morphology with substantial rearrangement of the actin cytoskeleton, as shown by staining with rhodamine phalloidin for visualization of actin filaments (Figure 4). Indeed, the APLP2-positive cells tend to display high levels of cortical actin, whereas the cells with APLP2 expression knocked down have less cortical actin and more intracellular microfilaments (Figure 4).

Bottom Line: Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine.Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade.Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA.

ABSTRACT
Amyloid precursor-like protein 2 (APLP2) is aberrantly expressed in pancreatic cancer. Here we showed that APLP2 is increased in pancreatic cancer metastases, particularly in metastatic lesions found in the diaphragm and intestine. Examination of matched human primary tumor-liver metastasis pairs showed that 38.1% of the patients had positive APLP2 expression in both the primary tumor and the corresponding liver metastasis. Stable knock-down of APLP2 expression (with inducible shRNA) in pancreatic cancer cells reduced the ability of these cells to migrate and invade. Loss of APLP2 decreased cortical actin and increased intracellular actin filaments in pancreatic cancer cells. Down-regulation of APLP2 decreased the weight and metastasis of orthotopically transplanted pancreatic tumors in nude mice.

Show MeSH
Related in: MedlinePlus