Adjacent Healthy Tissues Research Articles

Study of hot stress dynamic IR thermography for detecting surface cancerous tissue

The early diagnosis of cancer at any location of the human body can help in enhancing the survival rate and in reducing the cost of the treatment. Among the several techniques, the conventional infra-red (IR) thermography suffers from several limitations, including higher patient discomfort and longer time for tumour detection based on the temperature difference between tumour and adjacent healthy tissues. Hence, in this work, a novel non-invasive hot stress dynamic IR thermography system is proposed to detect surface tumour without severe discomfort to the patient. The system is designed to detect the surface tumour under 3 min within a temperature range of 42 and 37 °C. A two-dimensional numerical model based on the bio-heat transfer of tissue consists of cancerous and healthy cells is developed and validated to analysis the thermal contrast arises due to the presence of cancerous and healthy cells while cooling naturally. A light source is introduced with appropriate intensity to achieve a suitable temperature contrast. Moreover, the effects of natural convective heat loss from the tissue to the ambient and the scanning speed of IR Thermography on the tissue are investigated. A temperature difference of about 1.5 °C is found after cooling of tissues for 140 s, which can be detected using a thermographic camera. Finally, a sensitivity study is conducted to access the importance of the individual parameter over the final temperature field. The results predict the blood perfusion rate as the most significant parameter that significantly influences the temperature distribution in the considered domain.

Association of Abl interactor 2, ABI2, with platelet/lymphocyte ratio in patients with renal cell carcinoma: A pilot study.

There are many unknown aspects of the pathogenesis of renal cell carcinoma (RCC). The aim of the current study was to define new RCC-related genes and measure their associations with RCC and clinical parameters, especially platelet/lymphocyte ratio which may be an independent predictor of prognosis in patients with RCC and other forms of cancer. Via in silico analysis upon RCC-specific deleted genes in chromosome 3, four possible ceRNAs (ATXN3, ABI2, GOLGB1 and SMAD2) were identified. Then, the expression levels of these genes in tumour and adjacent healthy kidney tissues of 19 RCC patients were determined by real-time PCR. ATXN3 and GOLGB1 gene expression levels increased but ABI2 gene expression level decreased in tumour kidney tissues when compared to normal ones. ATXN3, ABI2 and GOLGB1 gene expression levels were significantly higher in Fuhrman grade 4 than other grades (P<.001). ABI2 gene expression levels were significantly associated with higher platelet/lymphocyte ratio of the patients with RCC (P<.05). ATXN3, ABI2 and GOLGB1 may predict higher RCC grades. Also, ABI2 may regulate platelet/lymphocyte ratio which may be an independent predictor of RCC and other forms of cancer.

Investigation of GHSR methylation levels in thymomas from patients with Myasthenia Gravis

Hypermethylation of the growth hormone secretagogue receptor gene (GHSR) is increasingly observed in human cancers, suggesting that it could represent a pan-cancer biomarker of clinical interest. However, little is still known concerning GHSR methylation levels in thymic epithelial tumors, and particularly in thymomas from patients with Myasthenia Gravis (TAMG). In the present study we collected DNA samples from circulating lymphocytes and surgically resected tumor tissues of 65 TAMG patients, and from the adjacent healthy thymic tissue available from 43 of them. We then investigated GHSR methylation levels in the collected tissues searching for correlation with the clinical characteristics of the samples. GHSR hypermethylation was observed in 18 thymoma samples (28%) compared to the healthy thymic tissues (P<1×10-4), and those samples were particularly enriched in advanced disease stages than stage I (94% were in stage II or higher). GHSR was demethylated in the remaining 47 thymomas, as well as in all the investigated healthy thymic samples and in circulating lymphocytes. GHSR hypermethylation is not a pan-cancer marker or an early event in TAMG, but occurs in almost 1/4 of them and mainly from stage II onward. Subsequent studies are required to clarify the molecular pathways leading to GHSR hypermethylation in TAMG tissues and their relevance to disease progression.

Tissue stiffness contributes to YAP activation in bladder cancer patients undergoing transurethral resection.

Changes in the cellular microenvironment play a critical role in the development of bladder cancer (BC). Yes-associated protein (YAP), a central mediator of the Hippo pathway, functions as a nuclear sensor of mechanotransduction that can be induced by stiffness of the extracellular matrix (ECM), including stiffness resulting from surgical manipulations. We aimed to clarify the possible association between surgically-related ECM stiffness and YAP activation in BC patients. We compared 30 bladder cancer tissues with grade II (n = 15 recurrent and n = 15 newly diagnosed) with 30 adjacent healthy tissues. Atomic force microscopy showed that patients with recurrent BC had stiffer ECM than newly diagnosed patients (P < 0.05). Gene expression profiles showed that β1 integrin (ITGB1), focal adhesion kinase (FAK), CDC42, and YAP were upregulated in cancerous tissues (P < 0.05); additionally, β1 integrin activation was confirmed using a specific antibody. Nuclear localization of YAP was higher in recurrent cancerous tissues compared with newly diagnosed and it was positively associated with higher stiffness (P < 0.05). Our results suggest that postsurgery-induced ECM stiffness can influence integrin-FAK-YAP activity and thereby YAP trafficking to the nucleus where it contributes to BC progression and relapse.

Bioinformatics analysis of key biomarkers and potential molecular mechanisms in hepatocellular carcinoma induced by hepatitis B virus.

Background:Hepatocellular carcinoma (HCC) accounts for up to 90% of all primary hepatic malignancies; it is the sixth most common cancer and the second most common cause of cancer mortality worldwide. Numerous studies have shown that hepatitis B virus and its products, HBV integration, and mutation can induce HCC. However, the molecular mechanisms underpinning the regulation of HCC induced by HBV remain unclear.Methods:We downloaded 2 gene expression profiling datasets, of HBV and of HCC induced by HBV, from the gene expression omnibus (GEO) database. Differentially expressed genes (DEGs) between HCC and HBV were identified to explore any predisposing changes in gene expression associated with HCC. DEGs between HCC and adjacent healthy tissues were investigated to identify genes that may play a key role in HCC. Any overlapping genes among these DEGs were included in our bioinformatics analysis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of overlapping genes were performed using the Metascape online database; the protein–protein interaction (PPI) network was analyzed using the STRING online database; and we obtained the hub genes of the PPI network using Cytoscape software. An overall survival (OS) analysis of hub genes was performed using km-plotter and the gene expression profiling interactive analysis (GEPIA) online database. The expression levels of hub genes were determined using the TCGA and GEPIA databases. Finally, the relationships between hub genes and tumors were analyzed using the comparative toxicogenomics database (CTD).Results:We identified 113 overlapping genes from the 2 datasets. Using functional and pathway analyses, we found that the overlapping genes were mainly related to the AMPK signaling pathway and cellular responses to cadmium ions. C8A, SPP2, KLKB1, PROZ, C6, FETUB, MBL2, HGFAC, C8B, and ANGPTL3 were identified as hub genes and C8A, SPP2, PROZ, C6, HGFAC, and C8B were found to be significant for survival.Conclusion:The DEGs re-analyzed between HCC and hepatitis B enable a systematic understanding of the molecular mechanisms of HCC reliant on hepatitis B virus.

Three-Dimensionally Printed On-Skin Radiation Shields Using High-Density Filament

PurposeCustom-fabricated lead shields are often used for superficial radiation treatments to reduce radiation doses to adjacent healthy tissue. However, the process for fabricating these lead shields is time consuming, labor intensive, and uncomfortable for patients. Alternatively, patient-specific shields can be 3-dimensionally (3D) printed from a high-density bronze-based filament to address these concerns. This study was performed to assess the shielding characteristics of 3D-printed bronze (3DPB) shields, demonstrate their clinical viability, and report the first ever published case of a patient treated with a 3DPB shield. Methods and MaterialsThe transmission of 6 and 9 MeV electron beams through varying thicknesses of 3DPB was first measured. Percent depth doses and beam profiles were measured with flat 3DPB shields and equivalent lead shields to determine surface dose enhancement, output factors, and field widths. Two 3DPB shields were designed and fabricated for an anthropomorphic phantom, and phantom measurements were performed using optically stimulated luminescence dosimeters and film. Finally, 3DPB shields were used during the treatment of 7 patients’ skin lesions. ResultsTen and 15 mm of 3DPB were sufficient to shield 6 and 9 MeV electrons, respectively, by 95%. The 3DPB and lead shields had nearly identical beam widths (within 1%). Output factors were on average within 0.8% for bronze shields and 1.2% for lead shields relative to an unshielded field. The skin enhancement for bronze was higher than for lead by an average of 6.3%. Phantom measurements using 3DPB shields generally showed less than 3% transmission of the primary beam under the 3DPB shield. The patients’ shields fit as designed and were all deemed clinically acceptable by their physicians. ConclusionsThe 3DPB shields fit better than lead shields, are easier to design and manufacture, and have similar dosimetric properties. 3DPB shields are a viable clinical option for patient-specific superficial shielding.

Lack of association between LincRNA-Pou3f gene expression and clinicopathological features in gastric cancer tissue

Emerging evidence supports the effective therapeutics for advanced or metastatic gastric cancers which have undergone little progress. Multiple pathophysiological process of lincRNA-POU3F3 and the critical roles in gastric cancer are not clearly defined. The aim of this study was to evaluate the expression of LincRNA-POU3F3 in gastric cancer compared to the adjacent healthy tissue to gain further knowledge about the aberrantly expressed value of selected lincRNA. The RNA was extracted from 42 gastric tumor tissue samples and paired adjacent normal tissues from the patients included in the present study. The expression of LincRNA-POU3F3 was assessed using the Real-time polymerase chain reaction. B2M was used as an internal control. The 2−ΔΔCq method was used to determine the expression fold changes (tumor vs. adjacent normal tissue). The extent of the LincRNA-POU3F3 gene expression was compared among 42 tumor tissues and adjacent normal tissues (the mean age of the study group was 60.32 ± 14.185 years). The level of the LincRNA-POU3F3 gene expression revealed no statistically significant difference between the tumor and adjacent normal tissues (P = 0.065). In order to further evaluate the role of LincRNA-POU3F3 in gastric cancer, the associations between the transcript levels of the gene and several clinicopathological features demonstrated no significant difference between the stages I, III and IV (P = 0.84), tumor grade groups (P = 0.44), Helicobacter pylori (H. pylori) infection (P = 0.06) and tumor size (P = 0.71). Taken together, our results demonstrated that the expression of LincRNA-POU3F3 at the transcriptional level had no significant difference in gastric cancer tissue samples. Due to this, it is not a potential target for diagnostic/therapeutic purposes.

MiR‑133b affects cell proliferation, invasion and chemosensitivity in renal cell carcinoma by inhibiting the ERK signaling pathway

Renal cell carcinoma has the highest incidence rate of cancer types in the urinary system. Moreover, microRNAs (miRNA) have been closely associated with numerous types of tumor. The present study aimed to investigate the effects of miRNA (miR)-133b on the proliferation, invasion and chemosensitivity of renal cell carcinoma cells, and to determine whether its mechanism was regulated by the ERK signaling pathway. Both renal cell carcinoma and adjacent healthy tissues from 60 patients, in addition to renal cell carcinoma lines, ACHN, Caki-1, A-498 and 786-O, and 293 cells, were used in this study. miR-133b expression was measured from renal cell carcinoma, adjacent healthy tissues and renal cell carcinoma cell lines by reverse transcription-quantitative PCR. Cells were transfected with miR-133b mimic to achieve miR-133b overexpression. The proliferative, migratory and invasive ability of the cells were evaluated using MTT, wound healing and Matrigel assays, respectively, and flow cytometry was used to detect the apoptotic rate. Following treatment with an ERK inhibitor, U0126, and activator, LM22B-10, western blotting was used to detect the expression of related proteins and the activity of the ERK signaling pathway. The overexpression of miR-133b significantly inhibited cell proliferation, migration and invasion, whilst inducing apoptosis and increasing the drug sensitivity of renal cell carcinoma cells to cisplatin, docetaxel and doxorubicin. The miR-133b mimic also increased the protein expression levels of Bax and decreased the expression levels of matrix metalloproteinase (MMP)-2, MMP-9, ATP-binding cassette subfamily G2, P-glycoprotein, Bcl-2 and proliferating cell nuclear antigen, as well as the phosphorylation of ERK (P<0.05). The administration of the U0216 inhibitor demonstrated similar effects to miR-133b overexpression, and there was no significant difference compared with the miR-133b mimic transfection (P>0.05). However, the overexpression of miR-133b combined with LM22B-10 treatment weakened the anticancer effects of miR-133b mimic transfection (P<0.05). In conclusion, miR-133b overexpression was observed to inhibit the proliferation, migration and invasion of renal cell carcinoma cells and improve chemotherapeutic sensitivity; it was suggested that the mechanism maybe related to the inhibition of ERK1/2 phosphorylation and thus decreased ERK signaling pathway activity.

Multi-dimensional analysis of tumor-specific and bystander CD8+ tumor infiltrating lymphocytes with TetTCR-SeqHD

Abstract Objective Antigen specific CD8+ T cells were isolated from primary tumor tissue and analyzed for on the single cell level to determine tumor-specific signatures for further study. Methods Primary tumor, metastatic lesions, adjacent healthy tissue and peripheral blood were from collected from patients at Dell Seton Medical Center at the University of Texas. Matched tumor and healthy samples underwent exome and RNA sequencing to allow prediction of tumor-specific epitopes. We then synthesized DNA-barcoded tetramers for all predicted tumor associated-epitopes and an HLA-matched library of exogenous epitopes. Tetramer stained CD8+ T cells were then sorted and analyzed by TetTCR-SeqHD, a technology that we recently developed to link TCR antigen specificity, TCR sequence, gene expression, and phenotype of single T cells. Results We analyzed ~2900 tumor infiltrating lymphocytes (TILs) and T cells from adjacent healthy tissue and observed enrichment for cytotoxic effector and tissue resident memory cells within the tumor versus the healthy tissues. We also note an enrichment of bystander cells, specifically those reactive to Epstein Barr Virus epitopes, within the cytotoxic cluster. Finally, we analyze the distribution and differential expression between tumor-specific lymphocytes and bystander T cells, noting reduced expression of CD57 and CD49d in the tumor-specific TILs, as previously described. Conclusions We validate the use of TetTCR-SeqHD for gaining insight into phenotypic and gene expression differences between bystander and tumor-specific TILs. We identified markers for future exploration and demonstrate key differences between shared clonal populations within healthy and cancerous tissues.

Downregulation of lncRNA PMS2L2 in patients with gastric adenocarcinoma predicts poor prognosis.

Long non-coding RNA PMS1 homolog 2 mismatch repair system component pseudogene 2 (PMS2L2) is a key player in lipopolysaccharide-induced inflammatory responses. Preliminary deep sequencing data revealed that PMS2L2 was downregulated in gastric adenocarcinoma (GA) tissues compared with healthy adjacent tissues and the aim of the present study was to investigate the role of PMS2L2 in GA. In the present study, reverse transcription-quantitative PCR assays were performed to analyze gene expression. Cell transfections were performed to analyze gene interactions and Transwell assays were performed to analyze cell invasion and migration. The results revealed that PMS2L2 expression was downregulated in cancer tissues obtained from patients with GA compared with healthy adjacent tissues and was not significantly affected by clinical stage. Furthermore, low levels of PMS2L2 in cancer tissues were closely associated with a low overall 5-year survival rate in patients. MicroRNA (miR)-25 was upregulated in GA tissues compared with healthy adjacent tissues and inversely associated with PMS2L2 levels. In GA cells in vitro, overexpression of PMS2L2 downregulated the expression of miR-25, while miR-25 overexpression did not significantly affect PMS2L2 expression. Furthermore, PMS2L2 overexpression inhibited the migration and invasion of GA cells. miR-25 overexpression partially rescued the decreased migration and invasion of GA cells caused by PMS2L2 overexpression. Therefore, PMS2L2 may downregulate miR-25 expression to inhibit GA.

Gene Therapy Repairs for the Epileptic Brain: Potential for Treatment and Future Directions.

Epilepsy is a syndrome specified by frequent seizures and is one of the most prevalent neurological conditions, and that one-third of people of epilepsy are resistant to available drugs. Surgery is supposed to be the main treatment for the remedy of multiple drug-resistant epilepsy, but it is a drastic procedure. Advancement in genomic technologies indicates that gene therapy can make such surgery unnecessary. The considerable number of new studies show the significance of mutation in mammalian target of rapamycin pathway, NMDA receptors, GABA receptors, potassium channels and G-protein coupled receptors. Illustration of the meticulous drug in epilepsy targeting new expression of mutations in SCN8A, GRIN2A, GRIN2D and KCNT1 are conferred. Various methods are utilized to express a gene in a precise area of the brain; Transplantation of cells in an ex vivo approach (fetal cells, fibroblasts, immortalized cells), nonviral vector delivery and viral vector delivery like retrovirus, herpes simplex virus adenovirus and adeno-related virus. Gene therapy has thus been explored to generate anti-epileptogenic, anti-seizure and disease-modifying effects. Specific targeting of the epileptogenic region is facilitated by gene therapy, hence sparing the adjacent healthy tissue and decreasing the adverse effects that frequently go hand in hand with antiepileptic medication.

The Intelligent Knife: New Approach to Cancer Surgery

The idealized goal of cancer surgery is to resect all cancerous tissue and no adjacent healthy tissue. To achieve this goal, physicians rely on their

Overexpression of antisense long non‑coding RNA ZNF710‑AS1‑202 promotes cell proliferation and inhibits apoptosis of clear cell renal cell carcinoma via regulation of ZNF710 expression.

Antisense long non-coding RNAs (AS lncRNAs) have been increasingly recognized as important regulators of gene expression and have been found to play crucial roles in the development and progression of tumors. The present study explored the roles of AS lncRNA ZNF710-AS1-202 in clear cell renal cell carcinoma (ccRCC). The expression levels of ZNF710-AS1-202 were detected in 46 human ccRCC tissues and 34 healthy adjacent renal tissues. The associations between the levels of ZNF710-AS1-202 expression and the clinicopathological features of the patients were evaluated by the χ2 test. Gain- and loss-of-function experiments were performed to analyze the role of ZNF710-AS1-202 in ccRCC cell proliferation and survival in vitro. Reverse transcription-quantitative PCR and/or western blotting were employed to detect ZNF710-AS1-202, zinc finger protein 710 (ZNF710) and cyclin B1 expression. The Cell Counting Kit-8 and colony formation assays, as well as flow cytometry, were used to detect cell proliferation or apoptosis. The subcellular localization of ZNF710-AS1-202 was analyzed by RNA fluorescence in situ hybridization. The results revealed that ZNF710-AS1-202 was downregulated in human ccRCC tissues and was associated with the pathological grade, tumor size, local invasion and TNM stage, but not with lymph node metastasis or distant metastasis. However, ZNF710-AS1-202 overexpression promoted the proliferation of RCC cells and inhibited apoptosis. Opposite results were observed when ZNF710-AS1-202 was knocked down by small interfering RNA. Furthermore, ZNF710-AS1-202, which was mainly expressed in the cytoplasm of RCC cells, regulated ZNF710 mRNA and protein expression in opposing manners. In conclusion, the present study revealed that ZNF710-AS1-202 and ZNF710 may serve as promising therapeutic targets for ccRCC.

LINC00978 promotes hepatocellular carcinoma carcinogenesis partly via activating the MAPK/ERK pathway.

Objective: To study the role of long non-coding RNA (lncRNA) LINC00978 in hepatocellular carcinoma (HCC) carcinogenesis.Materials and methods: LINC00978 expression level was measured by reverse transcription quantitative real-time PCR (RT-qPCR) in HCC tissues and adjacent healthy liver tissues from 49 HCC patients. MTT assay, colony forming assay, and flow cytometry were performed to evaluate the effects of shRNA-mediated LINC00978 knockdown on HCC cell proliferation, cell cycle progression, and apoptosis in vitro. Xenograft tumor model was performed to determine the effects of LINC00978 knockdown on HCC tumor growth in vivo. Western blot was used to assess the activation of signaling molecules in the apoptosis and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway.Results: LINC00978 expression was significantly up-regulated in human HCC tissue relative to adjacent normal tissue, and LINC00978 high expression was correlated with poor HCC overall survival. LINC00978 was up-regulated in HCC cell lines. ShRNA-mediated LINC00978 knockdown significantly decreased HCC cell proliferation, and induced HCC cell cycle arrest and apoptosis in vitro. LINC00978 knockdown led to significant decrease in tumor xenograft size in vivo. Western blots revealed LINC00978 inhibition decreased ERK, p38, and c-Jun N-terminal kinase (JNK) phosphorylation in HCC cells.Conclusions: LINC00978 is highly expressed in human HCC tissue and correlates with poor HCC prognosis. LINC00978 promotes HCC cell proliferation, cell cycle progression, and survival, partially by activating the MAPK/ERK pathway. Our findings partially elucidated the roles of LINC00978 in HCC carcinogenesis, and identified a therapeutic target for HCC.

Biophysicochemical motifs in T cell receptor sequences as a potential biomarker for high-grade serous ovarian carcinoma.

We previously showed, in a pilot study with publicly available data, that T cell receptor (TCR) repertoires from tumor infiltrating lymphocytes (TILs) could be distinguished from adjacent healthy tissue repertoires by the presence of TCRs bearing specific, biophysicochemical motifs in their antigen binding regions. We hypothesized that such motifs might allow development of a novel approach to cancer detection. The motifs were cancer specific and achieved high classification accuracy: we found distinct motifs for breast versus colorectal cancer-associated repertoires, and the colorectal cancer motif achieved 93% accuracy, while the breast cancer motif achieved 94% accuracy. In the current study, we sought to determine whether such motifs exist for ovarian cancer, a cancer type for which detection methods are urgently needed. We made two significant advances over the prior work. First, the prior study used patient-matched TILs and healthy repertoires, collecting healthy tissue adjacent to the tumors. The current study collected TILs from patients with high-grade serous ovarian carcinoma (HGSOC) and healthy ovary repertoires from cancer-free women undergoing hysterectomy/salpingo-oophorectomy for benign disease. Thus, the classification task is distinguishing women with cancer from women without cancer. Second, in the prior study, classification accuracy was measured by patient-hold-out cross-validation on the training data. In the current study, classification accuracy was additionally assessed on an independent cohort not used during model development to establish the generalizability of the motif to unseen data. Classification accuracy was 95% by patient-hold-out cross-validation on the training set and 80% when the model was applied to the blinded test set. The results on the blinded test set demonstrate a biophysicochemical TCR motif found overwhelmingly in women with HGSOC but rarely in women with healthy ovaries, strengthening the proposal that cancer detection approaches might benefit from incorporation of TCR motif-based biomarkers. Furthermore, these results call for studies on large cohorts to establish higher classification accuracies, as well as for studies in other cancer types.

A large air gap magnetic levitator for intra-arterial positioning of a clinical microcatheter: design, fabrication, and preliminary experimental evaluation

Radioembolization, a targeted treatment for advanced-stage liver cancer, is a medical procedure in which millions of radioactive microspheres (20–40 µm in diameter) are released into the blood vessels feeding liver tumors. These microspheres not only reduce blood flow to the tumors; their associated radiation kills the cancerous tissues into which they embed. While effective, this procedure also poses a threat to adjacent (non-target) healthy liver tissues. Recent research (both theoretical and experimental) has established a strong relationship between microsphere injection location (in the artery cross section) and the destination vessel, demonstrating the potential for specific vessel and tumor targeting. One such method for positioning the catheter tip, the subject of this research, is to use magnetic levitation. This paper (the second of a two-part series) details the design, fabrication, and preliminary testing of a large air gap magnetic levitator. This device, with a model-based sliding mode controller, is used to precisely position the microcatheter tip in a dynamically perfused arterial model cross section, allowing for microsphere injections from controlled locations within the cross section. Microsphere distributions were found to be significantly dependent on release location, and the large gap levitator is shown to be a viable option for catheter positioning.

LncRNA NCK1-AS1 Promotes Cancer Cell Proliferation and Increase Cell Stemness in Urinary Bladder Cancer Patients by Downregulating miR-143.

BackgroundLong noncoding RNAs (lncRNAs) play critical and complex roles in regulating various biological processes of cancers. Our study aimed to investigate the involvement of lncRNA NCK1-AS1 in urinary bladder cancer (UBC).MethodsqRT-PCR was used to detect the expression of lncRNA NCK1-AS1 and miR-143 in UBC tissues and cells. The dual-luciferase reporter system assays were used to confirm the interaction between NCK1-AS1 and miR-143, and flow cytometry assays were applied to examine the behavioral changes in HT-1376 and HT-1197 cell lines.ResultsIt was observed that NCK1-AS1 was up-regulated, while miR-143 was down-regulated in tumor tissues than in adjacent healthy tissues of urinary bladder cancer (UBC) patients. A 5-year survival analysis showed that the survival rate of patients with high NCK1-AS1 level or low miR-143 level in tumor tissues appears relatively low. Correlation analysis revealed a significant inverse correlation between NCK1-AS1 and miR-143 in tumor tissues. Over-expression NCK1-AS1 reduced the expression level of miR-143, while elevating the level of miR-143 failed to affect NCK1-AS1 expression. NCK1-AS1 over-expression led to promoted proliferation and increased percentage of CD133+ (stemness) cells.ConclusionTherefore, NCK1-AS1 promotes cancer cell proliferation and increases cell stemness in UBC patients by down-regulating miR-143.

The Disturbed Iron Phenotype of Tumor Cells and Macrophages in Renal Cell Carcinoma Influences Tumor Growth.

Accumulating evidence suggests that iron homeostasis is disturbed in tumors. We aimed at clarifying the distribution of iron in renal cell carcinoma (RCC). Considering the pivotal role of macrophages for iron homeostasis and their association with poor clinical outcome, we investigated the role of macrophage-secreted iron for tumor progression by applying a novel chelation approach. We applied flow cytometry and multiplex-immunohistochemistry to detect iron-dependent markers and analyzed iron distribution with atomic absorption spectrometry in patients diagnosed with RCC. We further analyzed the functional significance of iron by applying a novel extracellular chelator using RCC cell lines as well as patient-derived primary cells. The expression of iron-regulated genes was significantly elevated in tumors compared to adjacent healthy tissue. Iron retention was detected in tumor cells, whereas tumor-associated macrophages showed an iron-release phenotype accompanied by enhanced expression of ferroportin. We found increased iron amounts in extracellular fluids, which in turn stimulated tumor cell proliferation and migration. In vitro, macrophage-derived iron showed pro-tumor functions, whereas application of an extracellular chelator blocked these effects. Our study provides new insights in iron distribution and iron-handling in RCC. Chelators that specifically scavenge iron in the extracellular space confirmed the importance of macrophage-secreted iron in promoting tumor growth.

Investigation of MMP-9 rs3918242 and TIMP-2 rs8179090 polymorphisms in renal cell carcinoma tissues

Abstract Background The proteolytic activities of matrix metalloproteinases (MMP), cell surface enzymes degrading extracellular matrix, is inhibited by matrix metalloproteinase tissue inhibitors (TIMP). We aim to detect the effects of MMP-9 rs3918242 and TIMP-2 rs8179090 gene variations in renal cell cancer transformation. Methods One hundred tumor and 100 adjacent healthy tissues were obtained from the patients with renal cell cancer. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) was performed and the products carried out in agarose gel electrophoresis were visualized under UV light. Statistical analyses were performed using SPSS 22 and p-values of less than 0.05 were considered as statistically significant. Results MMP-9 rs3918242 T allele was higher in tumor tissues (36.5%) than adjacent tissues (17%) and odds ratio was found 2.8 in T allele (p&lt;0.001). Odds ratio values were found 3.325 in the carriers of TT genotype and 3.5 in the carriers of CT genotype compared to the carriers of CC genotype (p&lt;0.01). The polymorphism of TIMP-2 rs8179090 was not found statically significant in tumor and adjacent tissues (p&gt;0.05). Conclusion MMP-9 rs3918242 T allele, TT and CT genotypes can be used as biomarkers in determining of renal cell carcinoma.

Long non-coding RNA CCAT1 promotes colorectal cancer cell migration, invasiveness and viability by upregulating VEGF via negative modulation of microRNA-218.

Increasing evidence has demonstrated that long non-coding (lnc) RNA is aberrantly expressed in numerous types of cancer. Colorectal cancer is a common malignancy; however, the role and mechanism underlying the influence of lncRNA-colon cancer associated transcript 1 (CCAT1) in colorectal cancer is yet to be elucidated. The present study revealed that CCAT1 is highly expressed in colorectal cancer tissues. Bioinformatics analysis and a dual-luciferase reporter gene assay indicated that CCAT1 and microRNA (miR)-218 had complementary binding sites. Furthermore, reverse transcription-quantitative PCR revealed that miR-218 was downregulated in colorectal cancer tissues compared with paired adjacent healthy tissues. To investigate the biological effects of CCAT1 on colorectal cancer cells, MTT and Transwell assays were performed. The results revealed that when compared with the control group, CCAT1-short hairpin (sh)RNA significantly inhibited colorectal cancer cell (SW480) viability and decreased migration and invasiveness. In addition, CCAT1-shRNA significantly reduced vascular endothelial growth factor (VEGF) expression in SW480 cells; however, these effects were partially rescued by an miR-218 inhibitor. Furthermore, it was revealed that the CCAT1-plasmid significantly promoted the viability of SW480 cells, increased cell migration and invasiveness, and significantly increased VEGF expression. However, these effects were also partially rescued by with a miR-218 mimic. Taken together, the present results identified that the CCAT1/miR-218 axis serves a key role in the regulation of colorectal cancer progression, which may be used as potential therapeutic target for the treatment of colorectal cancer.