Dataset Information

Functional proteomics characterization of the role of SPRYD7 in colorectal cancer progression and metastasis

ABSTRACT: SPRY domain-containing protein 7 (SPRYD7) is a barely known protein identified by spatial proteomics as upregulated in highly-metastatic to liver KM12SM colorectal cancer (CRC) cells in comparison to its isogenic poorly-metastatic KM12C CRC cells. Here, we aimed at analyzing SPRYD7 role in CRC by functional proteomics. By immunohistochemistry, the overexpression of SPRYD7 was observed to be associated to poor survival of CRC patients, and to an aggressive and metastatic phenotype. SPRYD7 stably overexpression was performed in KM12C and SW480 poorly-metastatic CRC cells, and in their isogenic highly-metastatic to liver KM12SM and to lymph nodes SW620 CRC cells, respectively. After confirming the upregulation of SPRYD7, in vitro and in vivo functional assays confirmed a key role of SPRYD7 in proliferation and migration of CRC cells. Furthermore, SPRYD7 was observed as an inductor of angiogenesis. In addition, the dysregulated SPRYD7-associated proteome and SPRYD7 interactors were elucidated by 10-plex TMT quantitative proteins, immunoproteomics, and bioinformatics. After WB validation, the biological pathways associated to the stably overexpression of SPRYD7 were visualized. In conclusion, it was demonstrated here that SPRYD7 is a novel protein associated to CRC progression and metastasis. Thus, SPRYD7 and its interactors might be of relevance to identify novel therapeutic targets for advanced CRC.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Colon Cancer

SUBMITTER: Ana Montero Calle

LAB HEAD: Rodrigo Barderas

PROVIDER: PXD042874 | Pride | 2024-01-26

REPOSITORIES: Pride

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Functional Proteomics Characterization of the Role of SPRYD7 in Colorectal Cancer Progression and Metastasis.

Montero-Calle Ana A Jiménez de Ocaña Sofía S Benavente-Naranjo Ruth R Rejas-González Raquel R Bartolomé Rubén A RA Martínez-Useros Javier J Sanz Rodrigo R Dziaková Jana J Fernández-Aceñero María Jesús MJ Mendiola Marta M Casal José Ignacio JI Peláez-García Alberto A Barderas Rodrigo R

Cells 20231031 21

SPRY domain-containing protein 7 (SPRYD7) is a barely known protein identified via spatial proteomics as being upregulated in highly metastatic-to-liver KM12SM colorectal cancer (CRC) cells in comparison to its isogenic poorly metastatic KM12C CRC cells. Here, we aimed to analyze SPRYD7's role in CRC via functional proteomics. Through immunohistochemistry, the overexpression of SPRYD7 was observed to be associated with the poor survival of CRC patients and with an aggressive and metastatic pheno ...[more]

PMID: 37947626

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Project description:Colorectal cancer (CRC) is a genetically heterogeneous disease with several distinct morphological growth patterns. This study was aimed to investigate genes differentially expressed between ulcerative and polypoid colorectal CRC. cDNA microarray was first employed to compare the gene expression profiling of ulcerative and polypoid CRC with paired normal mucosa. Potential candidates identified by data filtering were further validated using quantitative real-time polymerase chain reaction, western blot and immunohistochemistry. Epigenetic regulation of gene expression was investigated using methylation-specific PCR (MSP). cDNA microarray identified 11 up-regulated and 14 down-regulated genes differentially expressed in both types of tumor compared to matched normal mucosa. Among them, S100P was the only upregulated gene preferentially associated with polypoid CRC (p = 0.032), whereas no genes demonstrated significantly differential association with ulcerative CRC. S100P protein and mRNA expression level of polypoid CRC was significantly higher than that of ulcerative CRC (p < 0.05, respectively). Immunoreactivity of S100P protein was localized predominantly in the nuclei and to a less extent in the cytoplasm. Overexpression of S100P occurred early in the adenoma stage. 80% (24/30) and 20% (6/30) polypoid CRC showed diffusely strong and moderate overexpression, respectively. In contrast, S100P was diffusely and strongly expressed in 15% (6/40) ulcerative CRC, with 52.5% (21/40) and 32.5% (13/40) tumors having moderate and weak overexpression, respectively. S100P overexpression was preferentially associated with polypoid CRC (p < 0.001). The relative methylation level determined by MSP was not statistically different between polypoid and ulcerative CRC (43.36% vs. 49.10%, p = 0.168), indicating that promoter hypomethylation was directly related to upregulation of S100P mRNA. The gene expression profiling of ulcerative and polypoid CRC with paired normal mucosa were compared. Dye swapping experiments with ulcerative CRC vs normal mucosa or polypoid CRC vs normal mucosa were performed and we averaged the log ratios of the duplicated spots on each slide.

			Action	DRS
	IP_SPRYD7_KM12C_RAW.zip	Other
	IP_SPRYD7_KM12C_SEARCH.zip	Other
	IP_SPRYD7_KM12SM_RAW.zip	Other
	IP_SPRYD7_KM12SM_SEARCH.zip	Other
	checksum.txt	Txt