Supplementary Table S3 PDF file 147K, List of primers

Supplementary Table S1 PDF file 125K, Immunohistochemical staining and FISH results of UHGPS

PDF file, 67K, Primer sequences in RT-PCR analysis.

PDF file, 93K, High expression of SPARCL1 is associated with survival of patients with colon cancers and related to prognosis in both early and late stages.

PDF file, 234K, Expression of SPARCL1 in RKO cells reduced tumor growth in xenograft mouse models.

PDF file, 89K, Standard of cytoplasm staining of SPARCL1.

PDF file - 111K, Methods: Includes Mammosphere formation assay, Cell transfection and reporter assays, RNAi studies, as well as Western blot analyses and xenograft data. Figures S1-S9 include the following: Marker expression in primary fibroblasts and BC cells (S1); Serial confocal images of representative mammospheres formed by PKH67-labeled BT474 cells in the presence or absence of CCL2 (S2); CCL2 expression in CAF3 cells upon CM treatments (S3); Sustained CCL2 induction in CAFs co-cultured with tumor cells (S4); Expression of CCR2 and CCR4 in BC cells (S5); HES1 and HEY1 reporter assays in BT474 cells (S6); In vitro secretase cleavage assay (S7); Representative IHC images of XP/CAF265922 xenograft tumors treated with PBS, IgG, or anti-CCL2 (S8); Representative IHC images of CCL2 and NOTCH1 in primary BC specimens (S9).

Supplementary Data from Antitumor Activity of Targeting Src Kinases in Endothelial and Myeloid Cell Compartments of the Tumor Microenvironment

PDF file, 65K, Determine the specificity of SPARCL1 antibody.

Supplementary Table Legend PDF file 58K, Legend for Supplementary Tables S1-S5

PDF file, 126K, Uni- and Multivariate COX analysis for SPARCL1 and overall survival of CRCs.

Supplementary Table S5 PDF file 57K, Relative gene copy number determined by qPCR analysis (ratio to adjacent normal tissue) of selected eight genes located on chromosome 12q13-15 of individual DDLS

Supplementary Data from Protein Kinase A Activation Confers Resistance to Trastuzumab in Human Breast Cancer Cell Lines

Supplementary Table S4 PDF file 59K, Relative gene copy number determined by qPCR analysis (ratio to adjacent normal tissue) of selected eight genes located on chromosome 12q13-15 of individual UHGPS

PDF file, 379K, Recombinant human SPARCL1 protein reversed malignant phenotypes colon cancer cells in vitro.

PDF file, 49K, The quality of CRC database in COH set and ZJU set.

Supplementary Figures S1-S2; Supplementary Tables S1-S3.

Abstract Circulating tumor cells (CTCs) which disseminate from the primary tumor site have been found to be a major source of metastatic tumor. However, CTCs are heterogeneous and most CTC DNA is fragmented due to apoptosis. To better understand the molecular pathogenesis of metastasis, it is imperative to obtain single, non-apoptotic CTCs and investigate the genetic concordance between primary tumor cells, CTCs and metastatic tumor cells. Due to the abundance of normal cells present in the blood and in tumor tissue, isolation of single tumor cells or pure cell populations of these cells is extremely difficult. The DEPArray platform is designated to collect single CTCs after Cell-Search enrichment, and to sort pure intact tumor cells from formalin-fixed, paraffin-embedded (FFPE) tissues. Taking advantage of this technology, we have isolated single CTCs from prostate cancer patients. Whole genome amplification was performed and genomes with high integrity were used to evaluate the hotspot mutations in PIK3CA. For comparison, we also made single cell suspensions from FFPE tissue blocks of primary tumors and metastatic tumor in lymph nodes from the same patients. Immunofluorescence staining for cytokeratin (CK) and vimentin (Vim) was performed, followed by isolation of pure tumor cells (CK+), stromal cells (Vim+) and double-positive cells (CK+/Vim+) from both primary tumor and lymph node samples. We plan to do genetic analysis on these cells and compare the results to determine the genetic concordance between primary tumor cells, CTCs and metastatic tumor cells. By using the DEPArray, we have successfully isolated single CTCs, and populations of tumor cells, stromal cells and double-positive cells from primary and metastatic tumors. We will present new data on the genetic concordance between these cells, which will provide new insight into our understanding of metastasis in prostate cancer. Citation Format: Lixin Yang, Linling Chen, Yafan Wang, Jeremy Jones, Yun Yen, Sofia Loera, Raju Pillai, Peiguo Chu, Dennis Weisenburger. Characterization of genetic concordance between primary tumor cells, circulating tumor cells, and metastatic tumor cells from patients with prostate cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4880. doi:10.1158/1538-7445.AM2015-4880

Background: Ribonucleotide reductase (RR) is an essential enzyme involved in DNA synthesis. We hypothesized that RR subunit M2 (RRM2) might be a novel prognostic and predictive biomarker for estrogen receptor (ER)-negative breast cancers.Methods: Individual and pooled survival analyses were conducted on six independent large-scale breast cancer microarray data sets; and findings were validated on a human breast tissue set (ZJU set).Results: Gene set enrichment analysis revealed that RRM2-high breast cancers were significantly enriched for expression of gene sets that increased in proliferation, invasiveness, undifferentiation, embryonic stem/progenitor-like phenotypes, and poor patient survival (p < 0.01). Independent and pooled analyses verified that increased RRM2 mRNA levels were associated with poor patient outcome in a dose-dependent manner. The prognostic power of RRM2 mRNA was comparable to multiple gene signatures, and it was superior to TNM stage. In ER-negative breast cancers, RRM2 showed more prognostic power than that in ER-positive breast cancers. Further analysis indicated that RRM2 was a more accurate prognostic biomarker for ER-negative breast cancers than the pathoclinical indicators and uPA. A new RR inhibitor, COH29, could significantly enhance the chemosensitivity to doxorubicin in ER-negative MDA-MB-231 cells, but not in ER-positive MCF-7 cells.Conclusion: RRM2 appears to be a promising prognostic biomarker and therapeutic target for ER-negative breast cancer patients.