SPARCL1 Expression Research Articles

Expression and prognosis effect of methylation-regulated SLIT3 and SPARCL1 genes in smoking-related lung adenocarcinoma

Objective: To investigate the expression and prognosis effect of methylation-regulated SLIT3 and SPRCL1 genes in smoking-related lung adenocarcinoma. Methods: The expression levels of SLIT3 and SPARCL1 in cigarette smoke-induced malignant transformed cell (S30) and lung adenocarcinoma (LUAD) cell lines were measured by real-time fluorescence quantitative PCR (qPCR). Datasets of mRNA expression, DNA methylation and patient information data were obtained from The Cancer Genome Altas (TCGA) database. The mRNA expression levels of SLIT3 and SPARCL1 were validated in LUAD tissues. The 10-year survival curve of LUAD patients with different smoking history was plotted, and the correlation between mRNA expression level and DNA methylation level of LUAD patients was further analyzed. S30 cells were treated with 5-azacytidine (5-aza), an inhibitor of DNA methyltransferase, to analyze the methylation regulatory mechanism of SLIT3 and SPRCL1. Results: The qPCR results showed the significant down-regulation of SLIT3 and SPARCL1 in S30 cell and four LUAD cell lines (SLIT3: 0.493±0.134 and 0.041±0.014, 0.161±0.023, 0.277±0.055, 0.035±0.005; SPARCL1: 0.507±0.131 and 0.453±0.045, 0.420±0.040, 0.153±0.035, 0.430±0.050; all P<0.01). Bioinformatics analysis showed that SLIT3 and SPARCL1 were low expressed in LUAD tissue (8.12±1.58 vs 10.84±0.69 and 11.46±1.06 vs 13.57±0.67; both P<0.001) compared with adjacent peritumoral tissues, and expression levels of SLIT3 and SPARCL1 were significantly correlated with smoking history (both P<0.001). Non-smoker with high expression of SLIT3 and SPARCL1 was associated with better prognosis among LUAD patients. There was a significant negative correlation between promoter methylation and mRNA expression level of the two genes (r=-0.208, -0.574; both P<0.001). 5-aza treatment significantly up-regulated the expression levels of SLIT3 and SPARCL1 genes in S30 cells (2.137±0.281, 3.657±0.882; both P<0.01). Conclusion: SLIT3 and SPARCL1 can be regulated by DNA methylation and down-regulated in LUAD tissue, which has important prognostic significance on the smoking-induced LUAD patients.

Decreased expression of cell adhesion genes in cancer stem-like cells isolated from primary oral squamous cell carcinomas.

The goal of this study was to isolate cancer stem-like cells marked by high expression of CD44, a putative cancer stem cell marker, from primary oral squamous cell carcinomas and identify distinctive gene expression patterns in these cells. From 1 October 2013 to 4 September 2015, 76 stage III-IV primary oral squamous cell carcinoma of the gingivobuccal sulcus were resected. In all, 13 tumours were analysed by immunohistochemistry to visualise CD44-expressing cells. Expression of CD44 within The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma RNA-sequencing data was also assessed. Seventy resected tumours were dissociated into single cells and stained with antibodies to CD44 as well as CD45 and CD31 (together referred as Lineage/Lin). From 45 of these, CD44+Lin- and CD44-Lin- subpopulations were successfully isolated using fluorescence-activated cell sorting, and good-quality RNA was obtained from 14 such sorted pairs. Libraries from five pairs were sequenced and the results analysed using bioinformatics tools. Reverse transcription quantitative polymerase chain reaction was performed to experimentally validate the differential expression of selected candidate genes identified from the transcriptome sequencing in the same 5 and an additional 9 tumours. CD44 was expressed on the surface of poorly differentiated tumour cells, and within the The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma samples, its messenger RNA levels were higher in tumours compared to normal. Transcriptomics revealed that 102 genes were upregulated and 85 genes were downregulated in CD44+Lin- compared to CD44-Lin- cells in at least 3 of the 5 tumours sequenced. The upregulated genes included those involved in immune regulation, while the downregulated genes were enriched for genes involved in cell adhesion. Decreased expression of PCDH18, MGP, SPARCL1 and KRTDAP was confirmed by reverse transcription quantitative polymerase chain reaction. Lower expression of the cell-cell adhesion molecule PCDH18 correlated with poorer overall survival in the The Cancer Genome Atlas-Head and Neck Squamous Cell Carcinoma data highlighting it as a potential negative prognostic factor in this cancer.

Integrated analysis reveals down-regulation of SPARCL1 is correlated with cervical cancer development and progression.

Cervical cancer is the fourth most common malignancy among women worldwide, and continued research to discover biomarkers or therapeutic targets will aid early diagnosis and treatment of this cancer. Here, we investigated novel cervical cancer biomarkers using integrated analysis of high-throughput sequencing data from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. We have identified nine genes of interest that appear to be involved in cervical cancer development: SPARCL1, SYCP2, KIF4A, PRC1, TOP2A, LAMP3, KIF20A, MCM2, and APOBEC3B. Furthermore, gene ontology (GO) and co-expression analysis of these differentially expressed genes indicated that SPARCL1 may play a core role in cervical cancer development. Further, we analyzed the expression of these nine genes during the progression of cervical cancer, and found that SPARCL1 is also related to precancerous lesions and migration processes during cervical cancer pathogenesis. Finally, we validated these observations by investigating SPARCL1 expression in cervical cancer tissue and serum samples. The diagnostic specificity of serum SPARCL1 in cervical cancer occurrence was also compared with other high incidence diseases. All of these data indicate that SPARCL1 may be a novel cancer predictive marker and a potential therapeutic target for tumor development and progression in cervical cancer.

SPARCL1 ACCELERATES SYMPTOM ONSET IN ALZHEIMER’S DISEASE AND INFLUENCES BRAIN STRUCTURE AND FUNCTION DURING AGING

Recent evidence indicates that the expression of SPARCL1, an astrocyte-secreted synaptogenic protein, is dysregulated in Alzheimer's disease (AD). However, the precise relationship between altered SPARCL1 expression and AD pathogenesis remains unclear. Here, we examined associations between single nucleotide polymorphisms (SNPs) in the SPARCL1 gene and several AD-related endophenotypes, including rates of brain atrophy, trajectories of cognitive performance, and longitudinal changes in resting-state regional cerebral blood flow (rCBF), in the Baltimore Longitudinal Study of Aging (BLSA). We first identified SPARCL1 SNPs associated with altered gene expression of SPARCL1 in the brain (GTEx: http://www.gtexportal.org/home/). We selected two variants, rs9998212 and rs7695558, that are associated with reduced gene expression in the hippocampus and frontal cortex, respectively (Broad Institute SNP Annotation and Proxy Search: http://archive.broadinstitute.org/mpg/snap/ldsearchpw.php). In the BLSA neuroimaging substudy (BLSA-NI), we compared longitudinal changes in brain volumes (MRI; n=120 participants; 56–85 years old; number of scans= 826) and rCBF (measured by 15O - water PET; n=81 participants; 56–85 years old; number of scans= 664) in cognitively normal participants between carriers and non-carriers of the SPARCL1 minor alleles. We then analyzed longitudinal changes in cognitive performance of 591 participants who remained cognitively normal (mean age at first assessment: 61 years; range: 45–93 years) and 129 subjects who eventually developed MCI or AD (mean age at first assessment: 73 years; range: 59–89 years), comparing minor allele carriers versus non-carriers. Both SPARCL1 SNPs were associated with longitudinal changes in rCBF in several brain regions, including those vulnerable to AD pathology (e.g., inferior temporal cortex and precuneus). Furthermore, SPARCL1 SNP rs9998212 was associated with widespread and significantly faster rates of brain atrophy in cognitively normal individuals. Lastly, in participants who developed incident MCI or AD, SPARCL1 SNP rs7695558 was associated with faster memory decline prior to disease onset. These findings suggest that SPARCL1 may influence the early stages of AD pathogenesis through widespread effects on brain structure and function. These biological actions may be mediated through its role in synaptic repair during neuronal injury in AD.

Brain endothelial cell expression of SPARCL-1 is specific to chronic multiple sclerosis lesions and is regulated by inflammatory mediators in vitro.

Cell matrix modulating protein SPARCL-1 is highly expressed by astrocytes during CNS development and following acute CNS damage. Applying NanoLC-MS/MS to CSF of RRMS and SPMS patients, we identified SPARCL-1 as differentially expressed between these two stages of MS, suggesting a potential as CSF biomarker to differentiate RRMS from SPMS and a role in MS pathogenesis. This study examines the potential of SPARCL-1 as CSF biomarker discriminating RRMS from SPMS in three independent cohorts (n = 249), analyses its expression pattern in MS lesions (n = 26), and studies its regulation in cultured human brain microvasculature endothelial cells (BEC) after exposure to MS-relevant inflammatory mediators. SPARCL-1 expression in CSF was significantly higher in SPMS compared to RRMS in a Dutch cohort of 76 patients. This finding was not replicated in 2 additional cohorts of MS patients from Sweden (n = 81) and Switzerland (n = 92). In chronic MS lesions, but not active lesions or NAWM, a vessel expression pattern of SPARCL-1 was observed in addition to the expression by astrocytes. EC were found to express SPARCL-1 in chronic MS lesions, and SPARCL-1 expression was regulated by MS-relevant inflammatory mediators in cultured human BEC. Conflicting results of SPARCL-1's differential expression in CSF of three independent cohorts of RRMS and SPMS patients precludes its use as biomarker for disease progression. The expression of SPARCL-1 by BEC in chronic MS lesions together with its regulation by inflammatory mediators in vitro suggest a role for SPARCL-1 in MS neuropathology, possibly at the brain vascular level.

Matricellular protein SPARCL1 regulates tumor microenvironment-dependent endothelial cell heterogeneity in colorectal carcinoma.

Different tumor microenvironments (TMEs) induce stromal cell plasticity that affects tumorigenesis. The impact of TME-dependent heterogeneity of tumor endothelial cells (TECs) on tumorigenesis is unclear. Here, we isolated pure TECs from human colorectal carcinomas (CRCs) that exhibited TMEs with either improved (Th1-TME CRCs) or worse clinical prognosis (control-TME CRCs). Transcriptome analyses identified markedly different gene clusters that reflected the tumorigenic and angiogenic activities of the respective TMEs. The gene encoding the matricellular protein SPARCL1 was most strongly upregulated in Th1-TME TECs. It was also highly expressed in ECs in healthy colon tissues and Th1-TME CRCs but low in control-TME CRCs. In vitro, SPARCL1 expression was induced in confluent, quiescent ECs and functionally contributed to EC quiescence by inhibiting proliferation, migration, and sprouting, whereas siRNA-mediated knockdown increased sprouting. In human CRC tissues and mouse models, vessels with SPARCL1 expression were larger and more densely covered by mural cells. SPARCL1 secretion from quiescent ECs inhibited mural cell migration, which likely led to stabilized mural cell coverage of mature vessels. Together, these findings demonstrate TME-dependent intertumoral TEC heterogeneity in CRC. They further indicate that TEC heterogeneity is regulated by SPARCL1, which promotes the cell quiescence and vessel homeostasis contributing to the favorable prognoses associated with Th1-TME CRCs.

Expression and clinical significance of secreted protein acidic and rich in cysteine like 1 in gastric cancer

Objective To investigate the expression and clinical significance of secreted protein a- cidic and rich in cysteine like 1 （SPARCL1） in gastric cancer. Methods Immunohistochemistry （IHC） was used to detect SPARCL1 protein expression in 106 cases of gastric cancer tissues, normal mucosae and lymph nodes. Results The expression of SPARCL1 in normal mueosae was obviously higher than that in gastric cancer tissues and metastatic lymph nodes （P 〈 0. 01 ）, and the down-regnlated SPARCL1 was re- lated to serosa invasion, lymph node metastasis, lymph node ratio （LNR）, tumor location, remote metas- tasis, clinical stage and survival time （ P 〈 0.05 ）. Furthermore, SPARCL1 was an important independent prognosis factor （ P 〈 O. 05 ）. Conclusion Down-regulated SPARCL1 might paly an important role in oc- currence, progression, metastasis and prognosis of gastric cancer. Key words: Secreted protein acidic and rich in cysteine like 1 ; Gastric carcinoma; Prognosis

Bioinformatic analysis of chemokine (C-C motif) ligand 21 and SPARC-like protein 1 revealing their associations with drug resistance in ovarian cancer

Chemokine (C-C motif) ligand 21 (CCL21) and SPARC-like protein 1 (SPARCL1/MAST9/hevin/SC-1) are associated with various biological behavior in the development of cancers. Although the expression of CCL21 and SPARCL1 is downregulated in many solid tumors, their roles in ovarian cancer and their associations with drug resistance have rarely been studied. We performed a comprehensive bioinformatic analysis consisting of motif analysis, literature co-occurrence, gene/protein-gene/protein interaction network, protein-small molecule interaction network, and microRNAs enrichments which revealed that CCL21 and SPARCL1 directly or indirectly interact with a number of genes, proteins, small molecules and pathways associated with drug resistance in ovarian and other cancers. These results suggested that CCL21 and SPARCL1 may contribute to drug resistance in ovarian cancer. This study provided important information for further investigation of drug resistance-related functions of CCL21 and SPARCL1 in ovarian cancer.

SPARCL1: a potential molecule associated with tumor diagnosis, progression and prognosis of colorectal cancer

We investigated whether SPARCL1 played an essential role in tumor initiation, formation and progression of colorectal carcinomas. In this study, we examined expression of SPARCL1 protein in the normal colorectal mucosa, adjacent normal mucosa and primary and lymph node metastases from colorectal cancer patients. In matched patients, we found that SPARCL1 was negative in the distant normal colorectal mucosa, weakly expressed in the adjacent normal mucosa, strongly expressed in primary colorectal adenocarcinomas and slightly expressed in their lymph node metastases. A similar pattern was observed in the SPARCL1 expression from our series of non-matched colorectal cancer patients. The strongest expression and highest frequency of the SPARCL1 protein were found in the primary cancers. Interestingly, in the primary tumors, the frequency of SPARCL1 expression was significantly increased from the Dukes' A to Dukes' B tumors and then decreased gradually from the Dukes' B to C and D tumors. There was no difference in the intensity of SPARCL1 expression between the central areas and invasion margins of the primary tumors. Moreover, the SPARCL1 protein was more strongly expressed in the highly differentiated tumors than the lower differentiated ones. The patients with positive expression of SPARCL1 in their tumors had worse prognosis than the patients with SPARCL1-negative ones, even after the analyses by Multivariate and Interaction method. Expression of SPARCL1 protein might be a valuable biomarker for early diagnosis in colorectal cancers and further predicting patients' prognosis.

SPARCL1, Shp2, MSH2, E-cadherin, p53, ADCY-2 and MAPK are prognosis-related in colorectal cancer

To investigate the expression of markers that are correlated with the prognosis of colorectal cancer (CRC) patients. One hundred and fifty-six CRC patients were followed up for more than 3 years after radical surgery. Immunohistochemical (IHC) analysis was performed to detect the expression of 14 pathway-related markers (p53, APC, p21ras, E-cadherin, endothelin-B receptor, Shp2, ADCY-2, SPARCL1, neuroligin1, hsp27, mmp-9, MAPK, MSH2 and rho) in specimens from these patients. Bioinformatics analysis involving a Support Vector Machine (SVM) was used to determine the best prognostic model from combinations of these markers. Seven markers (SPARCL1, Shp2, MSH2, E-cadherin, p53, ADCY-2 and MAPK) were significantly related to the prognosis and clinical pathological features of the CRC patients (P < 0.05). Prognostic models were established through SVM from combinations of these 7 markers and proved able to differentiate patients with dissimilar survival, especially in stage II/III patients. According to the best prognostic model, the p53/SPARCL1 model, patients having high p53 and low SPARCL1 expression had about 50% lower 3-year survival than others (P < 0.001). SPARCL1, Shp2, MSH2, E-cadherin, p53, ADCY-2 and MAPK are potential prognostic markers in CRC. A p53/SPARCL1 bioinformatics model may be used as a supplement to tumor-nodes-metastasis staging.

Abstract 1741: Quantitative Real-Time PCR analysis of SPARCL1 and SPARC expression in colorectal cancer tissues

Abstract The SPARC family of proteins comprises 10 members, which share structural similarities in one or more protein domains. SPARC-like protein 1 (SPARCL1; also known as hevin) and SPARC are two important family members, which have been found to be involved in various tumors. The relationship between these two molecules and colorectal cancer and its liver metastasis has not yet been fully studied and understood. cDNA microarray was used to analyze the expression profiles of 22523 genes in CC531 rat colon adenocarcinoma cells. Briefly, 2 × 106 CC531 were injected into the portal vein of male Wag/Rij rats to grow in the liver. The tumor cells were then re-isolated from rat livers at different time points (after 3, 6, 9, 14 and 21 days). SPARCL 1 and SPARC were among the genes, which were up-regulated in the re-isolated metastatic cells as compared to control CC531 cells grown in vitro, especially on days 3 (two- and nine-fold for SPARCL1 and SPARC) and 6 (four- and eleven-fold for SPARCL1 and SPARC) following tumor implantation. These results motivated us to analyze the expression of these two genes in 65 colorectal cancer tissues taken from patients in different UICC tumor stages (I, n=11; II, n=23; III, n=17; IV, n=14). A mixture of 10 mucosa samples was taken for comparison as normal control. Quantitative real time-PCR results showed that SPARCL1 mRNA was significantly down-regulated in 44 samples (68%) with these levels being on average fourfold lower (0.24 ± 0.21) than those in normal mucosa. Normal levels were found in 18 samples (28%), and up-regulated levels were detected in only 3 samples (4%) with these levels being on average threefold higher than those in normal mucosa (3.0 ± 1.1). No correlation between this down- or up-regulation and tumor stage could be found. In contrast to the results for SPARCL1, SPARC showed an almost consistent up-regulation in 60 of the tumor samples (92%). The average level of SPARC up-regulation was higher than that of SPARCL1 (6.5 fold higher than in normal mucosa). It is worth noting here that in those 21 samples, in which SPARCL1 mRNA was not down-regulated, SPARC expression was on average twofold higher (13fold higher than in normal mucosa) than the average expression level of all samples. These results suggest a relationship between the two family members, but it is still not clear, whether they have opposing or complementary functions in the context of tumor initiation and progression. More studies should be performed to uncover the possible role of these proteins in the course of colorectal cancer and its liver metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1741.