Multiple Cancer Tissues Research Articles

Pathological and Biological Significance of the Specific Glycan, TRA-1-60, on Aggressive Gastric Adenocarcinoma

The glycans form a unique complex on the surface of cancer cells and play a pivotal role in tumor progression, impacting proliferation, invasion, and metastasis. TRA-1-60 is a glycan that was identified as a critical marker for the establishment of fully reprogrammed inducible pluripotent stem cells. Its expression has been detected in multiple cancer tissues, including embryonal carcinoma, prostate cancer, and pancreatic cancer, but the biological and pathological characterization of TRA-1-60-expressing tumor cells remains unclear within various types of malignancies. Here, we report the biological characteristics of TRA-1-60-expressing gastric cancer cells, especially those with its cell surface expression, and the therapeutic significance of targeting TRA-1-60. The cells with cell membrane expression of TRA-1-60 were mainly observed in the invasive area of patient gastric cancer tissues and correlated with advanced stages of the disease based on histopathological and clinicopathological analyses. In vitro analysis using a scirrhous gastric adenocarcinoma line, HSC-58, which highly expresses TRA-1-60 on its plasma membrane, revealed increased stress-resistant mechanisms, supported by the upregulation of glutathione synthetase and NCF-1 (p47phox) via lipid–ROS regulatory pathways, as detected by RNA-seq analysis followed by oxidative stress gene profiling. Our in vivo therapeutic study using the TRA-1-60-targeting antibody–drug conjugate, namely, Bstrongomab-conjugated monomethyl auristatin E, showed robust efficacy in a mouse model of peritoneal carcinomatosis induced by intraperitoneal xenograft of HSC-58, by markedly reducing massive tumor ascites. Thus, targeting the specific cell surface glycan, TRA-1-60, shows a significant therapeutic impact in advanced-stage gastric cancers.

Cellular and clinical impact of protein phosphatase enzyme epigenetic silencing in multiple cancer tissues

BackgroundProtein Phosphatase Enzymes (PPE) and protein kinases simultaneously control phosphorylation mechanisms that tightly regulate intracellular signalling pathways and stimulate cellular responses. In human malignancies, PPE and protein kinases are frequently mutated resulting in uncontrolled kinase activity and PPE suppression, leading to cell proliferation, migration and resistance to anti-cancer therapies. Cancer associated DNA hypermethylation at PPE promoters gives rise to transcriptional silencing (epimutations) and is a hallmark of cancer. Despite recent advances in sequencing technologies, data availability and computational capabilities, only a fraction of PPE have been reported as transcriptionally inactive as a consequence of epimutations.MethodsIn this study, we examined promoter-associated DNA methylation profiles in Protein Phosphatase Enzymes and their Interacting Proteins (PPEIP) in a cohort of 705 cancer patients in five tissues (Large intestine, Oesophagus, Lung, Pancreas and Stomach) in three cell models (primary tumours, cancer cell lines and 3D embedded cancer cell cultures). As a subset of PPEIP are known tumour suppressor genes, we analysed the impact of PPEIP promoter hypermethylation marks on gene expression, cellular networks and in a clinical setting.ResultsHere, we report epimutations in PPEIP are a frequent occurrence in the cancer genome and manifest independent of transcriptional activity. We observed that different tumours have varying susceptibility to epimutations and identify specific cellular signalling networks that are primarily affected by epimutations. Additionally, RNA-seq analysis showed the negative impact of epimutations on most (not all) Protein Tyrosine Phosphatase transcription. Finally, we detected novel clinical biomarkers that inform on patient mortality and anti-cancer treatment sensitivity.ConclusionsWe propose that DNA hypermethylation marks at PPEIP frequently contribute to the pathogenesis of malignancies and within the precision medicine space, hold promise as biomarkers to inform on clinical features such as patient survival and therapeutic response.

Integrating single-cell and bulk expression data to identify and analyze cancer prognosis-related genes

Compared with traditional evaluation methods of cancer prognosis based on tissue samples, single-cell sequencing technology can provide information on cell type heterogeneity for predicting biomarkers related to cancer prognosis. Therefore, the bulk and single-cell expression profiles of breast cancer and normal cells were comprehensively analyzed to identify malignant and non-malignant markers and construct a reliable prognosis model. We first screened highly reliable differentially expressed genes from bulk expression profiles of multiple breast cancer tissues and normal tissues, and inferred genes related to cell malignancy from single-cell data. Then we identified eight critical genes related to breast cancer to conduct Cox regression analysis, calculate polygenic risk score (PRS), and verify the predictive ability of PRS in two data groups. The results show that PRS can divide breast cancer patients into high-risk group and low-risk group. PRS is related to the overall survival time and relapse-free interval and is a prognosis factor independent of conventional clinicopathological characteristics. Breast cancer is usually regarded as a cancer with a relatively good prognosis. In order to further explore whether this workflow can be applied to cancer with poor prognosis, we selected lung cancer for a comparative study. The results show that this workflow can also build a reasonable prognosis model for lung cancer. This study provides new insight and practical source code for further research on cancer biomarkers and drug targets. It also provides basis for survival prediction, treatment response prediction, and personalized treatment.

Cell-in-cell: a potential biomarker of prognosis and a novel mechanism of drug resistance in cancer.

The cell-in-cell (CIC) phenomenon has received increasing attention over recent years because of its wide existence in multiple cancer tissues. The mechanism of CIC formation is considerably complex as it involves interactions between two cells. Although the molecular mechanisms of CIC formation have been extensively investigated, the process of CIC formation remains ambiguous. Currently, CIC is classified into four subtypes based on different cell types and inducing factors, and the underlying mechanisms for each subtype are distinct. Here, we investigated the subtypes of CIC and their major mechanisms involved in cancer development. To determine the clinical significance of CIC, we reviewed several clinical studies on CIC and found that CIC could serve as a diagnostic and prognostic biomarker. The implications of CIC on the clinical management of cancers also remain largely unknown. To clarify this aspect, in the present review, we highlight the findings of recent investigations on the causal link between CIC and cancer treatment. We also indicate the existing issues that need to be resolved urgently to provide a potential direction for future research on CIC.

Spatially resolved whole-transcriptome analysis with simultaneous highly multiplexed immune cell epitope detection in multiple cancer tissues

Abstract The tumor microenvironment is composed of highly heterogeneous niches, often with varying degrees of immune infiltration. The spatial distribution of immune cells with respect to malignant cells can directly impact patient prognosis and overall survival outcomes. The Visium CytAssist Spatial Gene Expression assay uses a whole transcriptome probe-based approach, termed RTL, to detect and quantify mRNA expression with spatial context. Although examination of the tumor microenvironment with an RTL-based spatial assay can provide significant transcriptomic information concerning regions of interest, immune cells frequently have extremely low mRNA expression levels and can be difficult to detect. The use of antibody-conjugated probes specific to immune cell epitopes, which are highly expressed, can enhance data recovered from these tumor samples, enabling spatially accurate detection of immune populations. The Visium CytAssist Spatial Proteogenomic Solution enables identification of immune-specific epitopes via antibody-conjugated probes from the same tissue slide used for transcriptomic analysis. Using the CytAssist workflow, we showcase the ability to comprehensively resolve immune cells associated with multiple immune and tumor tissues, including an array of human breast cancer punches. Spatial expression patterns of immune markers map back to distinct morphological features within the samples, allowing identification of differentially-expressed genes associated with those areas. Overall, these data highlight the value of Visium CytAssist Spatial Proteogenomic Solution in immuno-oncology studies, through the integration of spatially resolved transcriptomic and immune cell marker data.

Abstract 5643: High-spatial-resolution multi-omics analysis of cancer tissues

Abstract Recent technological breakthroughs in single cell sequencing have revealed that malignant cells and nonmalignant cells are both highly dynamic and in the case of tumor cells, that the intratumoral heterogeneity is quite remarkable. However, several questions remain, especially in a spatial context, and a rapidly growing need has been observed for imaging-based approaches with single-cell resolution to visualize and characterize the tumor microenvironment. In the current study, we used Miltenyi Biotec newly developed spatial gene expression technology and automated imaging platform, to acquire spatial microscopy data for multiple biological analytes and multiple cancer tissues. The approach is based on fluorescence microscopy and the MICS technology principle of cyclic staining (MACSima™ Imaging Cyclic Staining) along with validated fluorochrome-conjugated antibodies and specifically amplified oligonucleotide probes combined with a proprietary signal removal system. FFPE tissue blocks were analyzed and each gene, which display a unique emission spectrum during a single cycle, was decoded. Due to the non-destructive nature of the workflow, H&E staining, immunofluorescence and RNA data can be collected together into a single image and on a single section. Using our antibody and targeted RNA immune-oncology panel coupled with our smart segmentation and flexible gating software suite, we have generated a topographical map of the expression profile level for several cancer tissues i.e.: tonsil, pancreas, kidney and ovary. This technology allows for the analysis of hundreds of protein markers and RNA transcripts on a single tissue, the characterization of the spatial architecture and the expression topography of tumor tissue sections. A better understanding of tissue microenvironment and implementation of spatial biology approaches will help advance the identification of more suitable biomarkers and for cancer therapy. Citation Format: Emily Neil, Dongju Park, Erica Lloyd, Michael Dibuono, Seiyu Hosono, Reto Muller, Hanna Lafayette, Hsinyi Smith, Robert Pinard. High-spatial-resolution multi-omics analysis of cancer tissues. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5643.

The rise of viperin: the emerging role of viperin in cancer progression.

Viperin, an IFN-regulated gene product, is known to inhibit fatty acid β-oxidation in the mitochondria, which enhances glycolysis and lipogenesis during viral infections. Yet, its role in altering the phenotype of cancer cells has not been established. In this issue of the JCI, Choi, Kim, and co-authors report on a role of viperin in regulating metabolic alterations in cancer cells. The authors showed a correlation between clinical outcomes and viperin expression levels in multiple cancer tissues and proposed that viperin expression was upregulated in the tumor microenvironment via the JAK/STAT and PI3K/AKT/mTOR/HIF-1α pathways. Functionally, viperin increased lipogenesis and glycolysis in cancer cells by inhibiting fatty acid β-oxidation. Viperin expression also enhanced cancer stem cell properties, ultimately promoting tumor initiation in murine models. This study proposes a protumorigenic role for viperin and identifies HIF-1α as a transcription factor that increases viperin expression under serum starvation and hypoxia.

Inhibition of BRD4 enhanced the tumor suppression effect of dasatinib in gastric cancer

BRD4, a member of the bromodomain and extraterminal (BET) family, is elevated in multiple cancer tissues, including gastric cancer (GC). Targeted therapy with BRD4 may help improve the overall survival of patients with GC. Meanwhile, the approved multi-target kinase inhibitor, dasatinib, was recently reported to show varied tumor-suppressive effects in GC cells. This study investigated BRD4 expression in vivo and in vitro using immunohistochemistry and western blotting, respectively. We discussed the relationship between BRD4 expression and patient prognosis. Next, the antitumor efficacy of dasatinib was measured in BRD4-knockdown GC cells to determine the role of BRD4 blockage in dasatinib treatment. Finally, molibresib, a BET inhibitor, was used to measure the cooperative function of BRD4 inhibition and dasatinib treatment in three GC cell lines. Epithelial BRD4 expression was higher in tumoral and metastatic tissues and was strongly associated with unfavorable tumor, node, and metastasis stages and survival. BRD4 expression was heterogeneous in the three GC cell lines tested in vitro. In SGC7901, a BRD4-high GC cell line, knockdown of BRD4 using specific siRNAs suppressed cell growth individually and cooperatively with dasatinib. Moreover, molibresib and dasatinib showed a cooperative effect in suppressing the proliferation of BRD4-high GC cells. In conclusion, we confirmed that increased epithelial BRD4 expression is associated with poor disease stage and prognosis in GC and BRD4 blockage might be a valuable strategy to improve the sensitivity of dasatinib and other drugs in the chemotherapy of advanced GC.

Silencing of Circ_0135889 Restrains Proliferation and Tumorigenicity of Human Neuroblastoma Cells

IntroductionNeuroblastoma (NB) is the most common extracranial solid tumor in infants and young children. Circular ribonucleic acid (RNA) hsa_circ_0135889 (circ_0135889; hsa_circ argonaute 2 _001) is highly expressed in multiple cancer tissues, including NB. However, its role in tumor progression of NB was unclear. MethodsReal-time quantitative polymerace chain reaction was used to detect RNA expression, and western blotting, or immunohistochemistry was used to measure protein expression. Functional experiments were performed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, 5-Ethynyl-2’- deoxyuridine, Annexin V-fluorescein isothiocyanate/propidium iodide, and transwell assays, as well as xenograft tumor model. The intermolecular interaction was predicted by online databases and confirmed by dual-luciferase reporter assay and RNA pull-down assay. ResultsCirc_0135889 and neuronal differentiation 1 (NEUROD1) were upregulated whilst microRNA (miR)-127-5p, was downregulated in NB tumors and immortalized NB cells. Silencing of circ_0135889 could suppress cell proliferation, migration and invasion, but enhance apoptosis rate of NB cells in vitro. More importantly, circ_0135889 depletion inhibited xenograft tumor growth of NB cells. Circ_0135889 was a sponge for miR-127-5p, and inhibition of miR-127-5p counteracted the inhibitory impact of circ_0135889 knockdown on the malignant behaviors of NB cells. Moreover, NEUROD1 was a direct target of miR-127-5p, and miR-127-5p exerted the anti-tumor role in NB cells by targeting NEUROD1. Furthermore, circ_0135889 regulated NEUROD1expression by sponging miR-127-5p. ConclusionsCirc_0135889 promoted the tumorigenicity of NB by regulating miR-127-5p/NEUROD1 axis, which might provide a promising therapeutic target for NB.

Expression profiles of three proteins in various cancerous tissues of digestive system

A variety of gastrointestinal tumors are the most malignancies with high morbidity and mortality globally. The previous studies show that there are abnormal expressions of TATA-box binding protein associated factor 1 like (<italic>TAF</italic>1<italic>L</italic>), stomatin-like protein 1 (<italic>STOML</italic>1) and stomatin-like protein 2 (<italic>STOML</italic>2) genes in oral, esophageal and colorectal cancer lesions. To explore the potential roles of above three genes in the carcinogenesis of digestive system, the differential expressions of TAF1L, STOML1 and STOML2 in cancerous and paracancerous tissues are compared and analyzed by multiple immunohistochemical staining using microarray of 6 cancer tissues of esophagus, stomach, liver, pancreas, colon and rectum with matched paracancerous tissues. The results show that STOML2 is significantly overexpressed in all of 6 gastrointestinal cancers <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M4"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.001</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>, and STOML1, one of the same family member of STOML2, is highly expressed only in esophageal and liver cancers <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.005</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>. In addition, except for pancreatic cancer, TAF1L is also abnormal highly expressed in other 5 cancer tissues <inline-formula><mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"><mml:mo stretchy="false">(</mml:mo><mml:mi>P</mml:mi><mml:mo>&lt;</mml:mo><mml:mn mathvariant="normal">0.005</mml:mn><mml:mo stretchy="false">)</mml:mo></mml:math></inline-formula>. At the same lesion tissue section, STOML1, STOML2 and TAF1L proteins are colocated with the key protein MMP9 of epithelial-mesenchymal transformation in positive cells. This study indicates that the three proteins of TAF1L, STOML1 and STOML2 might have a wide range of cancer promoting effects in multiple cancer tissues in the digestive system.In addition, due to their different expression levels in a variety of cancer tissues, they have tissue specificity and different functional missions.Further exploration of the molecular pathological potential of TAF1L, STOML1 and STOML2 in the occurrence and development of malignant tumors in digestive system can make them serve as new molecular markers for early diagnosis, differential diagnosis, targeted therapy and prognosis prediction in multiple digestive tumors. <fig fig-type="abstract-image" id="F1" orientation="portrait" position="float"><graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="1000-2618-2022-39-3-253/E85B5A24-8ED1-441e-8E1E-AB53DA3D3AE9-F001.jpg"/></fig>

RPS13, a potential universal reference gene for normalisation of gene expression in multiple human normal and cancer tissue samples.

Reference genes are considered stable genes and are used for normalizing the gene expression profile across different cell types; as well as, in normal and diseased samples. However, these gene associates with different biological processes, and hence expression vary in different pathological conditions. Therefore, in the present study, eight different reference genes were used and compared to identify common reference gene usable for an array of different cell types and human cancers. The expression stability of the eight reference genes across eleven normal and cancerous tissues was confirmed through real time-qPCR. Ribosomal protein S13 (RPS13) was found to be a common and stable reference gene across intra- and inter-comparison between various normal and tumor tissue types. Further, TCGA data analysis across and between normal and tumor tissue types also showed minimum deviation in expression of RPS13 gene out of eight routinely used reference genes. RPS13 is the common stable reference gene in normalization for gene expression based analysis in cancer research.

243 PROGNOSTIC IMPACT OF CXCR7 AND CXCL12 EXPRESSION IN PATIENTS WITH ESOPHAGEAL ADENOCARCINOMA

Abstract Chemokines are major regulators of cell trafficking and adhesion. The chemokine CXCL12 and its receptors, CXCR4 and CXCR7, have recently been reported as biomarkers in various cancers, including esophageal squamous cell carcinoma; however, there are few studies of these chemokines in esophageal adenocarcinoma (EAC). In this study, we investigated the relationship between expression of CXCL12, CXCR4 and CXCR7, and prognosis in patients with EAC. Methods This study examined 55 patients with EAC who were treated in Toronto General Hospital from 2001 to 2010. Tissue microarray immunohistochemistry was used to evaluate the expression of CXCL12, CXCR4 and CXCR7. Evaluation of immunohistochemistry was performed by a pathologist without knowledge of patients’ information and scored based on a semiquantitative scoring system. The average score from multiple cancer tissues on the microarray was utilized and patients were divided into high or low expression groups using the median score as a cutoff point. These results were compared with the patients’ clinicopathological features and survival. Results The score of CXCR7 was positively correlated with that of CXCL12 (r = 0.3154). High CXCR7 expression was significantly associated with lymphatic invasion (present vs absent, P = 0.005), higher number of lymph node metastases (pN0–1 vs pN2–3, P = 0.0014) and TNM stage (Stage I-II vs III-IV, P = 0.0168). Patients with high CXCR7 (n = 23) expression was associated with worse overall (OS) and disease-free survival (DFS) (P = 0.0221, 0.0090, respectively), and patients with high CXCL12 (n = 24) tended to have worse OS and DFS (P = 0.1091, 0.1477, respectively). High expression of both CXCR7 and CXCL12 was an independent prognostic factor for DFS on multivariable analysis (HR0.3, 95%CI: 0.1–0.8, P = 0.0115). Conclusion High CXCR7 expression was associated with poor prognosis in patients with EAC, and high expression of CXCR7 and its ligand CXCL12, had a stronger association on prognosis. Further study of this potential biomarker using whole tissue samples and larger sample size is warranted.

CD8+ T-cell Immune Surveillance against a Tumor Antigen Encoded by the Oncogenic Long Noncoding RNA PVT1.

CD8+ T cells recognize peptides displayed by HLA class I molecules on cell surfaces, monitoring pathologic conditions such as cancer. Advances in proteogenomic analysis of HLA ligandomes have demonstrated that cells present a subset of cryptic peptides derived from noncoding regions of the genome; however, the roles of cryptic HLA ligands in tumor immunity remain unknown. In the current study, we comprehensively and quantitatively investigated the HLA class I ligandome of a set of human colorectal cancer and matched normal tissues, showing that cryptic translation products accounted for approximately 5% of the HLA class I ligandome. We also found that a peptide encoded by the long noncoding RNA (lncRNA) PVT1 was predominantly enriched in multiple colorectal cancer tissues. The PVT1 gene is located downstream of the MYC gene in the genome and is aberrantly overexpressed across a variety of cancers, reflecting its oncogenic property. The PVT1 peptide was recognized by patient CD8+ tumor-infiltrating lymphocytes, as well as peripheral blood mononuclear cells, suggesting the presence of patient immune surveillance. Our findings show that peptides can be translated from lncRNAs and presented by HLA class I and that cancer patient T cells are capable of sensing aberrations in noncoding regions of the genome.

SQSTM1/p62 regulate breast cancer progression and metastasis by inducing cell cycle arrest and regulating immune cell infiltration

The autophagy adaptor protein SQSTM1/p62 is overexpressed in breast cancer and has been identified as a metastasis-related protein. However, the mechanism by which SQSTM1/p62 contributes to breast cancer progression and tumor microenvironment remains unclear. This study revealed that silencing SQSTM1/p62 expression suppressed breast cancer progression via regulating cell proliferation and reshaping the tumor microenvironment (TME). Here, we found that SQSTM1/p62 was overexpressed in multiple human cancer tissue types and that was correlated with poor patient overall survival (OS) and disease-free survival (DFS). Moreover, we found that short-hairpin RNA (shRNA)-mediated knockdown of p62 expression significantly inhibited cell proliferation, migration, and invasion, and promoted cell death in vitro, as well as suppressed breast cancer growth and lung metastasis in vivo. In addition, flow cytometry analysis of splenocytes and tumor infiltrating lymphocytes (TILs) indicated that the numbers of CD8α+ interferon (IFN)-γ+ cells (CTLs) and CD4+IFN-γ+ (Th1) cells were increased while those of CD4+IL-4+ (Th2) cells, tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) were decreased. RT-PCR analyses showed that the gene expression of Th1/Th2 cytokines changed in the tumor microenvironment. Silencing SQSTM1/p62 suppressed tumor cell lung metastasis. Together, our results provide strong evidence that silencing tumor cell SQSTM1/p62 inhibited tumor growth and metastasis through cell cycle arrest and TME regulation. This finding provides a novel molecular therapeutic strategy for breast cancer progression and metastasis treatment.

Identification of Cysteine Protease Inhibitor CST2 as a Potential Biomarker for Colorectal Cancer.

Additional biomarkers for the development and progression of colorectal cancer (CRC) remain to be identified. Hence, the current study aimed to identify potential diagnostic markers for CRC. Analyses of cysteine protease inhibitor [cystatins (CSTs)] expression in CRC samples and its correlation with cancer stage or survival in patients with CRC demonstrated that CRC tissues had greater CST1 and CST2 mRNA expression compared to noncancerous adjacent tissues, while higher CST2 mRNA expression in CRC tissues was correlated with advanced stages and disease-free survival in patients with CRC, encouraging further exploration on the role of CST2 in CRC. Through an online database search and tissue microarray (TMA), we confirmed that CRC samples had higher CST2 expression compared to noncancerous adjacent tissue or normal colorectal tissues at both the mRNA and protein levels. TMA also revealed that colorectal adenoma, CRC, and metastatic CRC tissues exhibited a significantly increased CST2 protein expression. Accordingly, survival analysis demonstrated that the increase in CST2 protein expression was correlated with shorter overall survival of patients with CRC. Moreover, our results found a significant upregulation of CST2 in multiple cancer tissues. Taken together, these findings suggest the potential role of CST2 as a diagnostic and prognostic biomarker for CRC.

The Biogenesis, Functions, and Roles of circRNAs in Bladder Cancer.

Bladder cancer (BCa) is the 10th most prevalent malignancy worldwide and remains a crucial cause of cancer-related morbidity and mortality. Circular RNAs (circRNAs), a large class of endogenous non-coding RNAs, contain unique covalent closed structures and their biogenesis and turnover are regulated by multiple factors. Recently, multiple circRNAs have been found to serve as important factors in several biological processes such as tumorigenesis. An increasing amount of research discovered that circRNAs are dysregulated in multiple cancer tissues compared with matched normal tissues, especially in BCa, indicating that circRNAs can act as biomarkers for the diagnosis and prognosis of BCa. In this review, we focus on the biogenesis, properties, turnover, and functions of circRNAs, summarizing their potential functions and clinical implications in BCa.

Long Non-coding RNA Gas5 Is Associated With Preeclampsia and Regulates Biological Behaviors of Trophoblast via MicroRNA-21.

Preeclampsia is a lethal pregnancy specific hypertensive disorder involving multisystem. Despite extensive studies to investigate the causes of preeclampsia, the pathogenesis still remains largely unknown. Long non-coding RNAs (lncRNAs) are a diverse class of non-translated RNAs which play a crucial part in various biological phenomena. Although lncRNA Growth Arrest-Specific 5 (GAS5) aberrantly expressed in multiple cancer tissues and is implicated in multiple biological processes of tumor cells, little is known about its role in preeclampsia. In this study, 40 patients with preeclampsia and 32 gestational age matched normotension pregnant women were recruited. Using quantitative real-time polymerase chain reaction (qRT-PCR), we found higher expression of GAS5 in placenta of preclamsia affected women. The level of GAS5 existed strongly in correlation with Thrombin Time indicating coagulation function and other clinical parameters by Pearson correlation analysis. Then we constructed the GAS5 lentivirus expression vectors and transfected into human trophoblast cell lines HTR-8/SVneo and JEG-3. Using in vitro cell culture studies, we found an inhibited effect of GAS5 on proliferative ability, migratory ability and invasive ability however; no effect on apoptosis was detected. Further mechanistic analysis found that GAS5 modulated microRNA-21 (miR-21) in an opposite variation tendency by qRT-PCR and rescue experiment. In addition, inhibition of GAS5 promoted the activation of PI3K/AKT signaling pathway and its downstream proteins covering MMP-9 and TP53 as evident from our qRT-PCR and western blot analyses. Thus, we suggested that GAS5 might involve in pregnancy with preeclampsia by influencing the biological functions of trophoblast cells through the regulation of miR-21 and activation of PI3K/AKT signaling pathway and its downstream targets, which may contribute to reveal the nature of preeclampsia.

Expression of the NEK family in normal and cancer tissue: an immunohistochemical study

BackgroundThe NEK serine/threonine protein kinases are involved in cell cycle checkpoints, DNA damage repair, and apoptosis. Alterations in these pathways are frequently associated with cell malignant cellular transformations. Thyroid cancer is the most common malignant tumour in the endocrine system. Despite good treatment methods, the number of cases has increased significantly in recent years. Here, we studied the expression of NEK1, NEK2, NEK3, and NEK5 in different types of normal and malignant tissues, using tissue microarray analysis, and identified NEKs as potential markers in thyroid malignancy.MethodsThe studied cases comprised multiple cancer tissue microarrays, including breast, colon, esophagus, kidney, lung, pancreas, prostate, stomach, thyroid and uterine cervix, as well as 281 patients who underwent thyroid resection for thyroid cancer or thyroid nodules. The expression of NEK1, NEK2, NEK3, and NEK5 was analyzed by immunohistochemistry. The expression pattern was evaluated in terms of intensity by two methods, semiquantitative and quantitative, and was compared between normal and cancer tissue.ResultsWe analysed the expression of each member of the NEK family in a tissue-dependent manner. Compared to normal tissue, most of the evaluated proteins showed lower expression in lung tumour. However, in the thyroid, the expression was higher in malignant tissue, especially for NEK 1, NEK3 and NEK5. Concerning characteristics of the thyroid tumour, such as aggressiveness, NEK1 expression was higher in tumours with multifocality and in patients with lymph node metastasis. NEK3 expression was stronger in patients with stage II, that involved metastasis. NEK5, on the other hand, showed high expression in patients with invasion and metastasis and in patients with tumour size > 4 cm. Furthermore, this work, demonstrated for the first time a high specificity and sensitivity of over-expression of NEK1 in classical and follicular variants of papillary thyroid cancer and NEK3 in tall-cell papillary thyroid cancer.ConclusionTaken together, the NEK protein kinases emerge as important proteins in thyroid cancer development and may help to identify malignancy and aggressiveness features during diagnosis.Trial registrationThis study was retrospectively registered. www.accamargo.org.br/cientistas-pesquisadores/comite-de-etica-em-pequisa-cep.

Screening and surveillance of multiple solid tumours using plasma placental-like chondroitin sulfate A (pl-CSA)

Rationale: Placental-like chondroitin sulfate A (pl-CSA) is known to be exclusively synthesized in multiple cancer tissues and associated with disease severity. Here, we aimed to assess whether pl-CSA is released into bio-fluids and can serve as a cancer biomarker.Methods: A novel ELISA was developed to analyse pl-CSA content in bio-fluids using pl-CSA binding protein and an anti-pl-CSA antibody. Immunohistochemical staining of tissue chips was used as the gold standard control.Results: The developed ELISA method was specific and sensitive (1.22 μg/ml). The pl-CSA content was significantly higher in lysates and supernatants of cancer cell lines than in those of normal cell lines, in plasma from mouse cancer models than in that from control mice, and in plasma from patients with oesophageal, cervical, ovarian, or lung cancer than in that from healthy controls. Similar to the tissue chip analysis, which showed a significant difference in pl-CSA positivity between cancer tissues and normal adjacent tissues, the plasma pl-CSA analysis had 100% sensitivity and specificity for differentiating oesophageal and lung cancer patients from healthy controls. Importantly, in oesophageal and lung cancer patients, the pl-CSA content was significantly higher in late-stage disease than in early-stage disease, and it dramatically decreased after surgical resection of the tumour.Conclusion: These data indicate a direct link between plasma pl-CSA content and tumour presence, indicating that plasma pl-CSA may be a non-invasive biomarker with clinical applicability for the screening and surveillance of patients with multiple types of solid tumours.

ZMYND10, an epigenetically regulated tumor suppressor, exerts tumor-suppressive functions via miR145-5p/NEDD9 axis in breast cancer

BackgroundRecent studies suggested that ZMYND10 is a potential tumor suppressor gene in multiple tumor types. However, the mechanism by which ZMYND10 inhibits breast cancer remains unclear. Here, we investigated the role and mechanism of ZMYND10 in breast cancer inhibition.ResultsZMYND10 was dramatically reduced in multiple breast cancer cell lines and tissues, which was associated with promoter hypermethylation. Ectopic expression of ZMYND10 in silenced breast cancer cells induced cell apoptosis while suppressed cell growth, cell migration and invasion in vitro, and xenograft tumor growth in vivo. Furthermore, molecular mechanism studies indicated that ZMYND10 enhances expression of miR145-5p, which suppresses the expression of NEDD9 protein through directly targeting the 3'-untranslated region of NEDD9 mRNA.ConclusionsResults from this study show that ZMYND10 suppresses breast cancer tumorigenicity by inhibiting the miR145-5p/NEDD9 signaling pathway. This novel discovered signaling pathway may be a valid target for small molecules that might help to develop new therapies to better inhibit the breast cancer metastasis.