ABSTRACT
Signaling via epidermal growth factor receptor (EGFR) and Src kinase pathways promote triple-negative breast cancer (TNBC) cell invasion and tumor metastasis. Here, we address the role of Cdc42-interacting protein-4 (CIP4) in TNBC metastasis in vivo, and profile CIP4 expression in human breast cancer patients. In human TNBC cells, CIP4 knock-down (KD) led to less sustained activation of Erk kinase and impaired cell motility compared to control cells. This correlated with significant defects in 3D invasion of surrounding extracellular matrix by CIP4 KD TNBC cells when grown as spheroid colonies. In mammary orthotopic xenograft assays using both human TNBC cells (MDA-MB-231, HCC 1806) and rat MTLn3 cells, CIP4 silencing had no overt effect on tumor growth, but significantly reduced the incidence of lung metastases in each tumor model. In human invasive breast cancers, high CIP4 levels was significantly associated with high tumor stage, TNBC and HER2 subtypes, and risk of progression to metastatic disease. Together, these results implicate CIP4 in promoting metastasis in TNBCs.
Subject(s)
Microtubule-Associated Proteins/physiology , Triple Negative Breast Neoplasms/mortality , Triple Negative Breast Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Animals , Carcinoma, Ductal, Breast/diagnosis , Carcinoma, Ductal, Breast/genetics , Carcinoma, Ductal, Breast/mortality , Carcinoma, Ductal, Breast/pathology , Cell Line, Tumor , Female , Gene Expression Regulation, Neoplastic , Humans , Mice , Mice, Transgenic , Middle Aged , Minor Histocompatibility Antigens , Neoplasm Invasiveness , Neoplasm Metastasis , Prognosis , Rats , Triple Negative Breast Neoplasms/diagnosis , Triple Negative Breast Neoplasms/genetics , Xenograft Model Antitumor Assays , Young AdultABSTRACT
BACKGROUND: Intronic and intergenic long noncoding RNAs (lncRNAs) are emerging gene expression regulators. The molecular pathogenesis of renal cell carcinoma (RCC) is still poorly understood, and in particular, limited studies are available for intronic lncRNAs expressed in RCC. METHODS: Microarray experiments were performed with custom-designed arrays enriched with probes for lncRNAs mapping to intronic genomic regions. Samples from 18 primary RCC tumors and 11 nontumor adjacent matched tissues were analyzed. Meta-analyses were performed with microarray expression data from three additional human tissues (normal liver, prostate tumor and kidney nontumor samples), and with large-scale public data for epigenetic regulatory marks and for evolutionarily conserved sequences. RESULTS: A signature of 29 intronic lncRNAs differentially expressed between RCC and nontumor samples was obtained (false discovery rate (FDR) < 5%). A signature of 26 intronic lncRNAs significantly correlated with the RCC five-year patient survival outcome was identified (FDR < 5%, p-value ≤ 0.01). We identified 4303 intronic antisense lncRNAs expressed in RCC, of which 22% were significantly (p < 0.05) cis correlated with the expression of the mRNA in the same locus across RCC and three other human tissues. Gene Ontology (GO) analysis of those loci pointed to 'regulation of biological processes' as the main enriched category. A module map analysis of the protein-coding genes significantly (p < 0.05) trans correlated with the 20% most abundant lncRNAs, identified 51 enriched GO terms (p < 0.05). We determined that 60% of the expressed lncRNAs are evolutionarily conserved. At the genomic loci containing the intronic RCC-expressed lncRNAs, a strong association (p < 0.001) was found between their transcription start sites and genomic marks such as CpG islands, RNA Pol II binding and histones methylation and acetylation. CONCLUSION: Intronic antisense lncRNAs are widely expressed in RCC tumors. Some of them are significantly altered in RCC in comparison with nontumor samples. The majority of these lncRNAs is evolutionarily conserved and possibly modulated by epigenetic modifications. Our data suggest that these RCC lncRNAs may contribute to the complex network of regulatory RNAs playing a role in renal cell malignant transformation.
Subject(s)
Carcinoma, Renal Cell/metabolism , Kidney Neoplasms/metabolism , RNA, Long Noncoding/metabolism , Transcriptome , Base Sequence , Carcinoma, Renal Cell/genetics , Carcinoma, Renal Cell/mortality , Computer Simulation , Humans , Introns , Kidney Neoplasms/genetics , Kidney Neoplasms/mortality , Molecular Sequence Annotation , Molecular Sequence Data , Open Reading Frames , Organ Specificity , RNA, Long Noncoding/geneticsABSTRACT
OBJECTIVE: To investigate the expression of CASP7 protein in renal cell carcinoma clear cell subtype (ccRCC) and its value to predict cancer-specific survival (CSS). METHODS: A tissue microarray containing 120 samples of ccRCC, 45 non-ccRCC, and 66 nontumor paired samples from patients who underwent partial or radical nephrectomy was hybridized with anti-CASP7 antibody. Tissue sections were scored according to intensity and the percentage of stained cells. CASP7 immunostaining scores were used to estimate the association with clinicopathologic parameters and calculate Kaplan-Meier survival curves. RESULTS: Reduced CASP7 expression was observed in ccRCC and non-ccRCC subtypes in comparison with nontumor renal tissues (P <.0001). CASP7 immunostaining was associated (P <.05) with clinicopathologic parameters (size, incidental tumor, clinical stage, renal vein invasion, and tumor necrosis) and correlated with CSS (P = .032) and global survival (P = .046) of patients with ccRCC. In addition, CASP7 expression was able to substratify patients with ccRCC with favorable prognosis according to low clinical stage, in which negative CASP7 staining was associated with patients with lower CSS (P = .045). Finally, CASP7 staining was able to provide significant stratification according to CSS (P = .018) among patients with ccRCC with disease relapse. CONCLUSION: Our results implicate the loss of CASP7 expression in the aggressiveness of ccRCC and indicate its potential use as a clinical prognostic marker of the disease.
