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Heparan Sulfate Proteoglycans Mediate Renal Carcinoma Metastasis
Author(s) -
Qazi Henry,
Shi ZhongDong,
Song Jonathan W,
Cancel Limary M,
Munn Lance L,
Tarbell John M
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.808.6
Subject(s) - gene knockdown , metastasis , cancer research , clone (java method) , glycocalyx , heparan sulfate , gentamicin protection assay , biology , cell culture , chemistry , cancer , cell , pathology , microbiology and biotechnology , medicine , biochemistry , dna , genetics
The surface proteoglycan/glycoprotein layer (glycocalyx) on tumor cells has been associated with cellular functions that can potentially enable invasion and metastasis. In addition, aggressive tumor cells with high metastatic potential have enhanced invasion rates in response to interstitial flow stimuli in vitro . Our previous studies suggest that heparan sulfate (HS) in the glycocalyx plays an important role in this flow mediated mechanostransduction and upregulation of invasive and metastatic potential. In this study, highly metastatic renal cell carcinoma cells were genetically modified to suppress HS production by knocking down its synthetic enzyme NDST1. Using modified Boyden chamber and microfluidic assays, we show that flow‐enhanced invasion is suppressed in HS deficient cells. To assess the ability of these cells to metastasize in vivo , parental or knockdown cells expressing fluorescence reporters were injected into kidney capsules in SCID mice. Histological analysis confirmed that there was a large reduction (95%) in metastasis to distant organs by tumors formed from the NDST1 knockdown cells compared to control cells with intact HS. The ability of these cells to invade surrounding tissue was also impaired. The substantial inhibition of metastasis and invasion upon reduction of HS suggests an active role for the tumor cell glycocalyx in tumor progression. 1 Heparan sulfate knockdown induced reduction in invasive tumor colonies per tissue sample for both the single clone and combined clones models *Single Clone Model Combined Clones ModelControl Knockdown Δ% Control Knockdown Δ%Kidney 1.8 0.3 −86 0.8 † 0.0 −100 † Liver 1.7 0.1 −97 2.1 0.1 −94 Lungs 14.6 0.5 −97 7.2 0.1 −98 Abdomen 3.3 0.3 −92 2.6 0.0 −100 Spleen 1.8 0.0 −100 0.4 † 0.0 −100 †* p < 0.005 for each comparison between the number of invasive tumor colonies formed by either the control (CtGFP) or knockdown (shORG‐8) cell lines within each tissue type **† 0.005 < p < 0.05 for the samples in the combined clones models where there were fewer observed cases of invasive colonies formed by the control (CtGFP) cell line ** n=165 tissue samples for the single clone control case. n=66 tissue samples for the single clone knockdown case, and n=50 for the paired cases in the combined clones model