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Tumor Biology

, Volume 37, Issue 1, pp 591–599 | Cite as

PDGF-D promotes dermal fibroblast invasion in 3-dimensional extracellular matrix via Snail-mediated MT1-MMP upregulation

  • Zhuo Qin
  • Jinfa Feng
  • Yusi Liu
  • Li-Li Deng
  • Changlian Lu
Original Article

Abstract

Increasing attention has been focused on the malignant tumor microenvironment, which plays important roles in tumor occurrence, progression and metastasis. Fibroblasts are recruited by platelet-derived growth factor (PDGFs) and invade the tumor microenvironment. In the PDGF family, PDGF-B has been reported to play an important role in the recruitment and invasion programs. However, whether PDGF-D plays a role in these programs remains unclear. We generated a recombinant plasmid expressing human PDGF-D and transfected the plasmid to dermal fibroblasts to examine the effects on cell invasive activities in 3D type I collagen gels. PDGF-D plasmid transfection enhanced fibroblast invasive activities both in invasive cell numbers and invasion depth in 3D collagen gels. These effects were blocked by Snail-specific siRNA transfection. PDGF-D transfection significantly induced Snail expression at both mRNA and protein levels. PDGF-D further upregulated MT1-MMP mRNA and protein expressions and this was inhibited when Snail was knocked down by siRNA. Both Snail and MT1-MMP expressions in fibroblasts and cellular invasive activities in 3D collagen induced by PDGF-D were inhibited by LY294002, SP600125, and U1026, the inhibitors of PI3K, JNK, and ERK1/2 signaling pathways, respectively. However, no effects were observed in response to the P38MAPK signaling pathway inhibitor SB203580. These effects of PDGF-D were confirmed by using the culture supernatants of the transfectants. Taken together, these data demonstrate that PDGF-D plays important roles in the recruitment and invasion programs of fibroblasts via the activation of PI3K, JNK and ERK1/2 signaling pathways, and upregulation of Snail and downstream effecter MT1-MMP. These findings indicate that PDGF-D is an important player in the tumor microenvironment for fibroblast recruitment.

Keywords

PDGF-D Fibroblast Snail 3-dimensional extracellular matrix MT1-MMP Invasion 

Abbreviations

PDGF-D

Platelet-derived growth factor-D

3D

3-dimensional

ECM

Extracellular matrix

MT1-MMP

Membrane-type 1 matrix metalloproteinase

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Fetal bovine serum

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (81272376) and the Scientific Research Foundation for the Returned Overseas Chinese Scholars Heilongjiang Province (LC201010), and the Graduate Innovation Foundation (Harbin Medical University).

Supplementary material

13277_2015_3828_Fig5_ESM.gif (222 kb)
Supplement 1

PDGF-D enhanced human dermal fibroblast Snail and MT1-MMP expression. (A, D) Human dermal fibroblasts were transfected with PDGF-D plasmid (PDGF-D) or empty vector (Vect) for 24 h, RT-PCR analyses of Snail and MT1-MMP mRNA levels. (B, E) Western blot analysis of Snail and MT1-MMP protein after transfection as indicated. (C, F) Fibroblasts were incubated with the culture supernatant collected from the PDGF-D transfectants, Western blot analyses of Snail and MT1-MMP protein expression after 48 h or 72 h as indicated. (G) Fibroblasts were treated with inhibitors, JNK inhibitor SP600125 (SP), PI3K inhibitor LY294002 (LY), ERK1/2 inhibitor U1026 (U) or P38 inhibitor, SB203580 (SB), in the presence or absence of culture supernatant collected from the PDGF-D transfectants, Western blot analyses of Snail and MT1-MMP protein expression. * P < 0.05. (GIF 222 kb)

13277_2015_3828_MOESM1_ESM.tif (8.2 mb)
High resolution image (TIFF 8393 kb)
13277_2015_3828_Fig6_ESM.gif (160 kb)
Supplement 2

PDGF-D promotes rat dermal fibroblasts angiogenesis. (A) RT-PCR analyses of VEGF mRNA expression in rat dermal fibroblasts in the presence of PDGF-D.(B) Rat dermal fibroblasts or transfected PDGF-D plasmid cells were seeded on chick chorioallantoic membrane (CAM) for 3 days. After dehydration and immobilization, dark lines mark the CAM. White lines mark the cells, and Blue arrows mark the new blood vessel. Scale bar = 100 μm. *P < 0.05. (C) Rat dermal fibroblasts were transfected with scrambled siRNA or rat snail-specific siRNA in the presence of PDGFD, 24 h later, RT-PCR analyses of VEGF mRNA expression. *P < 0.05. (GIF 159 kb)

13277_2015_3828_MOESM2_ESM.tif (8.2 mb)
High resolution image (TIFF 4775 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Zhuo Qin
    • 1
  • Jinfa Feng
    • 2
  • Yusi Liu
    • 1
  • Li-Li Deng
    • 3
  • Changlian Lu
    • 1
  1. 1.Department of Biopharmaceutical Sciences, College of PharmacyHarbin Medical University HarbinHarbinPeople’s Republic of China
  2. 2.Department of General SurgeryHeilongjiang Province HospitalHarbinPeople’s Republic of China
  3. 3.Department of OncologyThe Second Affiliated Hospital of Harbin Medical UniversityHarbinPeople’s Republic of China

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