Tumor-free Tissue Research Articles

The biological process of lysine-tRNA charging is therapeutically targetable in liver cancer.

Background & AimsMature transfer RNAs (tRNA) charged with amino acids decode mRNA to synthesize proteins. Dysregulation of translational machineries has a fundamental impact on cancer biology. This study aims to map the tRNAome landscape in liver cancer patients and to explore potential therapeutic targets at the interface of charging amino acid with tRNA.MethodsResected tumour and paired tumour‐free (TFL) tissues from hepatocellular carcinoma (HCC) patients (n = 69), and healthy liver tissues from organ transplant donors (n = 21), HCC cell lines, and cholangiocarcinoma (CC) patient‐derived tumour organoids were used.ResultsThe expression levels of different mature tRNAs were highly correlated and closely clustered within individual tissues, suggesting that different members of the tRNAome function cooperatively in protein translation. Interestingly, high expression of tRNA‐Lys‐CUU in HCC tumours was associated with more tumour recurrence (HR 1.1; P = .022) and worse patient survival (HR 1.1; P = .0037). The expression of Lysyl‐tRNA Synthetase (KARS), the enzyme catalysing the charge of lysine to tRNA‐Lys‐CUU, was significantly upregulated in HCC tumour tissues compared to tumour‐free liver tissues. In HCC cell lines, lysine deprivation, KARS knockdown or treatment with the KARS inhibitor cladosporin effectively inhibited overall cell growth, single cell‐based colony formation and cell migration. This was mechanistically mediated by cell cycling arrest and induction of apoptosis. Finally, these inhibitory effects were confirmed in 3D cultured patient‐derived CC organoids.ConclusionsThe biological process of charging tRNA‐Lys‐CUU with lysine sustains liver cancer cell growth and migration, and is clinically relevant in HCC patients. This process can be therapeutically targeted and represents an unexplored territory for developing novel treatment strategies against liver cancer.

Factors contributing to delayed decannulation of temporary tracheostomies following free tissue reconstructive surgery for head and neck cancer

Temporary tracheostomies (TT) are performed to secure the airway perioperatively and postoperatively in head and neck cancer patients undergoing tumour resection and free tissue reconstructive surgery. Patients report that having a TT is unpleasant and they appreciate its removal at the earliest opportunity. Early removal not only improves patient satisfaction but should allow for a more rapid recovery. The aim of this prospective study was to assess factors that contribute to delays in decannulation following TT and hence to provide an insight into the factors that will support earlier decannulation when it is safe to do so. Consecutive patients who had TT over a six-month period were included. Delayed decannulation was defined as that after day seven postoperatively. There were 42 patients with a median (IQR) age of 70 (60-74) years, 26 of whom were men. The tracheostomy was surgical in 29 and percutaneous in 13. The median (IQR) time to decannulation was 4 (3-5) days (range 1-11 days). Seven patients had delayed removal (7-11 days), the reasons being hospital-acquired pneumonia (HAP) (n=4), prolonged stay in the high dependency unit (HDU) following postoperative myocardial infarction and cardiac arrest (n=1), failure to tolerate TT occlusion (n=1), and not stated (n=1). There were early postoperative complications in 14 patients but despite this seven decannulations were still performed within two and six days. Additional multiprofessional assessment over weekends is likely to facilitate earlier decannulation. As some TTs are removed after a few days there is a need for better selection to avoid their use in certain patients.

FcγRIIB engagement drives agonistic activity of Fc-engineered αOX40 antibody to stimulate human tumor-infiltrating T cells

BackgroundOX40 (CD134) is a costimulatory molecule of the tumor necrosis factor receptor superfamily that is currently being investigated as a target for cancer immunotherapy. However, despite promising results in murine...

Downregulation of TAP1 in Tumor-Free Tongue Contralateral to Squamous Cell Carcinoma of the Oral Tongue, an Indicator of Better Survival.

Oral cancers are surrounded by epithelium that histologically might seem normal, but genetically has aberrations. In patients with squamous cell carcinoma of the oral tongue (SCCOT), it is therefore important to study not only the tumor but also the clinically tumor-free contralateral tongue tissue that remains in the patient after treatment to map changes of prognostic and/or diagnostic value. The transporter associated with antigen processing (TAP) dimer is a key factor in the process of activating cytotoxic T cells. By downregulating the expression of TAP, tumor cells can escape cytotoxic T cell recognition. Biopsies from tumor and clinically tumor-free contralateral tongue tissue in 21 patients with SCCOT were analyzed together with tongue biopsies from 14 healthy individuals, which served as the control group. Dividing patients into TAP1-high and TAP1-low groups according to the median TAP1 level in tumor-free samples showed that patients with lower TAP1 mRNA levels in tumor-free samples had better overall (p = 0.003) and disease-free survival (p = 0.002). The results showing that TAP1 levels in tumor-free tongue tissue contralateral to the SCCOT correlate with survival is an important contribution to early diagnosis and follow up of SCCOT.

Overexpression of HOTAIR in Tumor Tissues of Patients with Colon Cancer Correlates with Tumor Metastasis and Differentiation

Background: Aberrant expression level of Hox transcript antisense intergenic RNA (HOTAIR) has been associated with the etiopathogenesis of numerous cancers. Studies on epidemiological data have demonstrated that the risk of susceptibility to colon cancer varies among different populations due to several reasons. In this study, we aimed to assess the expression level of HOTAIR in tumoral tissues of patients with colon cancer and compare it with normal marginal tissues. Methods: We recruited a total of 50 patients with colon cancer and collected tumoral and matched marginal tumor free tissues during surgery. Afterwards, we isolated the total RNA from each sample, synthesized cDNA, and performed quantitative analysis by Real-time PCR using the SYBR Green PCR Master Mix in order to measure the transcript level of HOTAIR in samples. Results: The expression level of HOTAIR was upregulated in tumor tissues compared with normal tumor-free marginal tissues belonging to colon cancer patients (P= 0.0023). Moreover, the expression level of HOTAIR and the clinicopathological specifications of the patients had statistically significant correlations. Conclusions: HOTAIR may play a role in the development of colon cancer and have the potential for application as a biomarker for colon cancer prognosis.

Increased miR-20b Level in High Grade Cervical Intraepithelial Neoplasia

Cervical cancer is a common malignant tumor worldwide ranking fourth in incidence and mortality among females, which was reduced significantly by cytology screening and human papilloma virus (HPV) DNA testing. The specificity of cytology is high; however, the sensitivity is low, in contrast to the HPV DNA testing. Despite the success of these measures, new biomarkers are still considered to aim increasing sensitivity and specificity of screening and diagnosis. Significant alterations in microRNA (miRNA) expression have been detected in several cancers with variable consistency. To investigate the stratification role of miRNAs between normal epithelium and cervical intraepithelial neoplasia (CIN2–3), we screened the expression of 667 miRNAs to identify significant markers (n = 10), out of them 9 miRNAs were applied in the study (miR-20b, −24, −26a, −29b, −99a, −100, −147, −212, −515-3p) along with RNU48 and U6 as the references. To benchmark the miRNAs, 22 paired (tumor-free and tumor tissue pairs) laser microdissection-obtained cervical formalin fixed, paraffin embedded tissue samples were assayed. The expression of miR-20b was 2.4 times higher in CIN2–3 samples as compared to normal tissues (p < 0.0001). In the HPV16-positive subsets of the samples (n = 13), miR-20b showed 2.9-times elevation (p < 0.001), whereas miR-515 was 1.15-times downregulated (p < 0.05) in CIN2–3 as compared to normal tissue. These results suggest the potential value of miR-20b as a statification biomarker in order to differentiate neoplastic and non-tumorous cases.

Levels of MUC1 in tumours and serum of patients with different sub-types of squamous cell carcinoma of the head and neck.

Mucin 1 (MUC1) is a membrane-bound and secreted glycoprotein that has a protective role in surface epithelia. We recently demonstrated that MUC1 mRNA expression was upregulated in tumour-free tongue tissues adjacent to squamous cell carcinoma of the oral tongue (SCCOT) compared with that in the tumour tissues. The present study investigated MUC1 protein in SCCOT tissue and serum from patients with squamous cell carcinoma of the head and neck (SCCHN) at different sub-sites. The results from immunohistochemistry demonstrated that all SCCOT tissues expressed MUC1; however, the protein levels were not correlated with MUC1 mRNA levels in the same tumours. Furthermore, serum MUC1 level was lower in patients with SCCOT, tonsil SCC and gingival SCC compared with that in healthy subjects; however, the difference was only significant for patients with SCCOT (P=0.0421). No correlation was seen between MUC1 level in tumour tissues and MUCI level in serum from the same patients. The absence of correlation between MUC1 protein and mRNA levels in SCCOT tissues emphasized the importance of validating genomic data in clinical samples. Although significant MUC1 downregulation was observed in the serum of patients with SCCOT, there was a large variation within the groups, suggesting that MUC1 may not be used as a biomarker for these types of tumors.

Evaluating of miR-184, miR-497, miR-378, miR-103 and miR-506 expression level in non-small cell lung cancer patients tissues compared with their normal marginal tissues

BackgroundLung cancer is one of the most common cancers worldwide whose prevalence and mortality have not any remarkable reduction in recent decades, in spite of the progression in screening and diagnostic methods. MiR-103, miR-184, miR-378, miR-497 and miR-506 are microRNAs which recent evidence has implied the possible roles of them in the carcinogenesis process. Hence, in this study we examined whether there are any significant alterations in the expression level of these micro RNAs during carcinogenesis procedure or not, and if we can use these factors as a diagnostic or prognostic biomarker for non-small cell lung cancer (NSCLC). MethodsFifty NSCLC patients participated in our study and tumor and adjacent normal tumor-free tissues were collected during the bronchoscopy. Total RNA was extracted from cells, cDNA was synthesized and then the expression level of target miRNAs was examined by quantitative Real-time PCR. Subsequently, expression levels of these genes and their correlation with clinicopathologic features of patients were analyzed with proper statistical tests. In these tests Pvalue <0.05 was considered significant. ResultsThe output data of our study showed the statistically significant down-expression of miR-103 (Fold change = 0.83, Pvalue = 0.036), miR-184a (Fold change = 0.52, Pvalue = 0.011), miR-378 (Fold change = 0.84, Pvalue = 0.041), and miR-497 (Fold change = 0.72, Pvalue = 0.021) between these micro-RNAs only mir-497 and miR-184a are potential biomarkers for NSCLC. Also, the expression levels of these miRNAs had significant relations with metastasis, lymph node involvement, and tumor cell differentiation degree and tumor size of NSCLC patients. Deregulated fifth microRNA, miR-506 was not statistically significant (Pvalue > 0.05). ConclusionAccording to the results of this study, 4 of our studied microRNAs have the potential to be used as diagnostic or prognostic biomarkers in case of confirmation of the future studies.

CONCORDANCE study: An observational, multicenter, prospective study to evaluate concordance of detecting EGFR mutations by circulating tumor-free DNA versus tissues biopsy in NSCLC.

e21615 Background: Epidermal growth factor receptor (EGFR) mutations on circulating tumour free DNA (ctDNA) by liquid biopsy is suitable option in those with difficulty in obtaining tissue samples. Correlation in tissue and plasma results of EFGRm has not been established in the Indian population. 1,2,3 This study was done to investigate the utility of ctDNA for EGFRm testing with Next-Generation Sequencing (NGS) in a real-world diagnostic setting. Methods: This is a multicentre, prospective, diagnostic observational study in 245 newly diagnosed treatment naïve, histologically confirmed, advanced lung adenocarcinoma patients. This was a single visit study. The primary objective of the study was to determine the level of concordance between EGFR mutation status obtained by tissue and ctDNA from blood (plasma) based testing in terms of overall concordance, sensitivity &amp; specificity. Study was registered at Clinicaltrials.gov NCT03562819 &amp; CTRI/2018/08/015290. Results: Seventy-five (30.6%) and eighty-four (34.3%) subjects showed positive mutation status by plasma &amp; tissue testing respectively. The overall concordance of 82.9% was observed between tissue and ctDNA (Plasma) based testing. Sensitivity of EGFR mutation status between ctDNA and tissue was observed for EGFR mutation subtypes was observed to be 100% while specificity was observed to be 90.1%. Plasma and tissue sample testing detected 1.2% (n = 3) and 2.4% (n = 6) positive in exon 20 T790M EGFR mutation, respectively. In terms of secondary outcomes, plasma sample testing detected 16.7% (n = 41) positive for Exon 19 deletions type EGFR mutation followed Exon 21 L858R [11.4% (n = 28)]. Conclusions: CONCORDANCE, a real-world study in Indian patients suggest that ctDNA testing for EGFR mutation analysis is a diagnostic option in newly diagnosed lung adenocarcinoma patients. EGFR Mutation testing in ctDNA, being non-invasive and especially in patients without available/evaluable tumor sample may enable more patients to receive appropriate targeted therapies. [Table: see text]

Aberrant expression of two miRNAs promotes proliferation, hepatitis B virus amplification, migration and invasion of hepatocellular carcinoma cells: evidence from bioinformatic analysis and experimental validation.

BackgroundAs key negative regulators of gene expression, microRNAs (miRNAs) play an important role in the onset and progression of hepatocellular carcinoma (HCC). This study aimed to identify the miRNAs involved in HCC carcinogenesis and their regulated genes.MethodsThe Gene Expression Omnibus (GEO) dataset (GSE108724) was chosen and explored to identify differentially expressed miRNAs using GEO2R. For the prediction of potential miRNA target genes, the miRTarBase was explored. Enrichment analysis of Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed by the DAVID online tool. The hub genes were screened out using the CytoHubba plug-in ranked by degrees. The networks between miRNAs and hub genes were constructed by Cytoscape software. MiRNA mimics and negative control were transfected into HCC cell lines and their effects on proliferation, hepatitis B virus DNA (HBV-DNA) replication, TP53 expression, migration, and invasion were investigated. The following methods were employed: MTT assay, quantitative PCR (qPCR) assay, western blotting, wound healing assay, and transwell assay.ResultsA total of 50 differentially expressed miRNAs were identified, including 20 upregulated and 30 downregulated miRNAs, in HCC tumor tissues compared to matched adjacent tumor-free tissues. The top three upregulated (miR-221-3p, miR-222-3p, and miR-18-5p) and downregulated (miR-375, miR-214-3p and miR-378d) miRNAs, ranked by |log2 fold change (log2FC)|, were chosen and their potential target genes were predicted. Two gene sets, targeted by the upregulated and the downregulated miRNAs, were identified respectively. GO and KEGG pathway analysis showed that the predicted target genes of upregulated and downregulated miRNAs were mainly enriched in the cell cycle and cancer-related pathways. The top ten hub nodes of gene sets ranked by degrees were identified as hub genes. Analysis of miRNA-hub gene network showed that miR-221-3p and miR-375 modulated most of the hub genes, especially involving regulation of TP53. The q-PCR results showed that miR-221-3p and miR-375 were markedly upregulated and downregulated, respectively, in HCC cells and HCC clinical tissue samples compared to non-tumoral tissues. Furthermore, miR-221-3p overexpression significantly enhanced proliferation, HBV-DNA replication, as well as the migration and invasion of HCC cells, whereas miR-375 overexpression resulted in opposite effects. Western blotting analysis showed that the overexpression of miR-221-3p and miR-375 reduced and increased TP53 expression, respectively.ConclusionThe present study revealed that miR-211-3p and miR-375 may exert vital effects on cell proliferation, HBV-DNA replication, cell migration, and invasion through the regulation of TP53 expression in HCC.

Abstract P1-10-28: Role of Sec62 in prediction of response to neoadjuvant chemotherapy in patients with primary breast cancer

Abstract Background Chromosome 3q26 amplifications have been shown to represent a frequent alteration in various cancer entities including breast cancer. SEC62 - a 3q26 encoded gene - was identified as a potential oncogene and tumor-driver-gene for the pathogenesis of breast cancer. Although the precise physiological function of the respective protein Sec62 is not completely understood, it is hypothesized, that Sec62 induces an increased stress tolerance and enhances cell migration in SEC62 overexpressing cells resulting in a high rate of lymphatic metastasis and poorer overall prognosis in tumor tissue with a high Sec62 expression. We sought to further elucidate the function of Sec62 in breast cancer with special focus on its predictive role on the response to neoadjuvant chemotherapy. Materials and Methods All patients treated for primary breast cancer with neoadjuvant chemotherapy at the Department of Gynaecology and Obstetrics, Saarland University Hospital, Homburg, Germany between 01/2007 and 12/2018 were enrolled in this study. The study was approved by the Saarland institutional review board. Sec62 protein levels were analyzed in tumor tissue samples using immunohistochemistry (IHC). Sec62 immunoreactivity was evaluated by three independent examiners using the immunoreactive score (IRS) according to Remmele and Stegner (0-12). For the assessment of Sec62 protein expression in tumor cells, we rated Sec62 “negative” 0-3, “low” for a score of 4-8 and “high” for 9-12 as described in previous publications. IHC was performed on initial breast core biopsy (CB) tissue (pre-treatment) and on definite pathology specimen (PS) obtained during breast surgery after completion of neoadjuvant chemotherapy. Sec62 expression in both samples were compared and correlated with response to neoadjuvant chemotherapy evaluated pathologically using the semi-quantitative response to neoadjuvant chemotherapy scoring system by Sinn et al. ranging from 0 (no effect) to 4 (no tumor detectable). Additionally, Sec62 expression in tumor tissue was compared with the histologically tumor-free tissue of the same patient. Sec62 expression levels and correlation with response to neoadjuvant chemotherapy were compared using Wilcoxon signed-rank, Mann-Whitney and Pearson`s chi square test. Results 203 patients were assessed for eligibility. Nine cases were excluded, due to incomplete clinical information or insufficient slide quality, leaving 194 patients for final analysis. Median patient age was 54 (22-86) years and median response to neoadjuvant chemotherapy score of 2 (0-4). 20 patients (10 %) had luminal A, 46 (24 %) luminal B, 60 (31 %) Her2/neu positive and 68 (35 %) triple negative breast cancer. All analyzed slides showed an over-expression of Sec62 in breast cancer cells but no positive staining of physiologic breast tissue cells. Median Sec62 expression in core biopsies (CB) (8 (2-12)) was significantly higher compared to expression in final specimen (PS) (4 (0-12); p ≤ 0.01). Median difference between Sec62 expression of CB and PS was 2 (0-4). Regarding response to neoadjuvant chemotherapy patients with a low Sec62 expression in PS and a difference ≥ 6 between CB and PS showed a significant higher median response score compared to other patients (2 (0-4) vs. 1 (0-2); p ≤ 0.01; 4 (1-4) vs. 1 (0-4); p ≤ 0.01). When looking at breast cancer subtypes these effects were strongest in Her2/positive and triple negative patients and median Sec62 expression showed the highest decrease between CB and PS in these two subgroups (6 (-3-12); 2 (-6-12)). Conclusion We identified Sec62 as a potential biomarker for prediction of response to neoadjuvant chemotherapy in patients with primary breast cancer. This effect was observed to be strongest in patients with Her2/positive and triple negative breast cancer. Citation Format: Julia Caroline Radosa, Mariz Kasoha, Merle Doerk, Barbara Linxweiler, Maximilian Linxweiler, Florian Bochen, Rainer M Bohle, Matthias Wagner, Marc P Radosa, Erich-Franz Solomayer, Zoltan Ferenc Takacs. Role of Sec62 in prediction of response to neoadjuvant chemotherapy in patients with primary breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P1-10-28.

Analysis of the methylation of CpG islands in the CDO1, TAC1 and CHFR genes in pancreatic ductal cancer.

No difference in the gene methylation status of tumor-suppression genes between pancreatic cancer tissues and adjacent non-cancer tissues is observed. The present study investigated whether the promoter CpG islands of the cysteine dioxygenase 1 (CDO1), tachykinin precursor 1 (TAC1) and checkpoint with forkhead and ring finger domains (CHFR) genes were methylated in pancreatic cancer and adjacent non-cancerous pancreatic tissue in order to determine if they could be considered as markers for the detection of pancreatic cancer. A total of 38 Formalin-fixed and paraffin-embedded pancreatic adenocarcinoma tissues and their adjacent non-cancerous specimens from patients with pancreatic cancer, as well as 9 non-cancerous pancreatic samples from patients without pancreatic adenocarcinoma were obtained following surgical resection. The hypermethylation of CpG islands was detected using a methylation-specific quantitative PCR. The methylation values were calculated using the ∆Cq method and were expressed as 2−ΔCq. The 2−ΔCq value of the CDO1 promoter from pancreatic adenocarcinoma specimens was significantly higher compared with that of adjacent non-cancerous and tumor-free pancreatic tissues (P<0.0001 and P=0.0008, respectively). The 2−ΔCq value of the TAC1 promoter of pancreatic adenocarcinoma was also significantly higher compared with that of adjacent non-cancerous tissues and tumor-free pancreatic samples (both P<0.0001). However, there was no significant difference in the 2−ΔCq value of the CHFR promoter among the pancreatic cancer, adjacent non-cancer tissue and tumor-free pancreatic samples. Furthermore, 12 out of the 38 pancreatic adenocarcinoma cases (31.6%) presented some methylation in the CHFR promoter. The results from Kaplan-Meier analysis between CHFR promoter methylation values and the clinicopathological characteristics of patients with pancreatic adenocarcinoma demonstrated that CHFR promoter methylation was significantly associated with lymph node metastasis. The methylation values of CDO1 and TAC1 promoters in cancer tissues were higher compared with adjacent tissues. However, whether hypermethylation of CDO1 and TAC1 promoters may serve as a biomarker in the diagnosis of pancreatic adenocarcinoma remains unclear.

Glioblastoma Exhibits Inter-Individual Heterogeneity of TSPO and LAT1 Expression in Neoplastic and Parenchymal Cells.

Molecular imaging is essential for diagnosis and treatment planning for glioblastoma patients. Positron emission tomography (PET) with tracers for the detection of the solute carrier family 7 member 5 (SLC7A5; also known as the amino acid transporter light chain L system, LAT1) and for the mitochondrial translocator protein (TSPO) is successfully used to provide additional information on tumor volume and prognosis. The current approaches for TSPO-PET and the visualization of tracer ([18F] Fluoroethyltyrosine, FET) uptake by LAT1 (FET-PET) do not yet exploit the full diagnostic potential of these molecular imaging techniques. Therefore, we investigated the expression of TSPO and LAT1 in patient glioblastoma (GBM) samples, as well as in various GBM mouse models representing patient GBMs of different genetic subtypes. By immunohistochemistry, we found that TSPO and LAT1 are upregulated in human GBM samples compared to normal brain tissue. Next, we orthotopically implanted patient-derived GBM cells, as well as genetically engineered murine GBM cells, representing different genetic subtypes of the disease. To determine TSPO and LAT1 expression, we performed immunofluorescence staining. We found that both TSPO and LAT1 expression was increased in tumor regions of the implanted human or murine GBM cells when compared to the neighboring mouse brain tissue. While LAT1 was largely restricted to tumor cells, we found that TSPO was also expressed by microglia, tumor-associated macrophages, endothelial cells, and pericytes. The Cancer Genome Atlas (TCGA)-data analysis corroborates the upregulation of TSPO in a bigger cohort of GBM patient samples compared to tumor-free brain tissue. In addition, AIF1 (the gene encoding for the myeloid cell marker Iba1) was also upregulated in GBM compared to the control. Interestingly, TSPO, as well as AIF1, showed significantly different expression levels depending on the GBM genetic subtype, with the highest expression being exhibited in the mesenchymal subtype. High TSPO and AIF1 expression also correlated with a significant decrease in patient survival compared to low expression. In line with this finding, the expression levels for TSPO and AIF1 were also significantly higher in (isocitrate-dehydrogenase wild-type) IDHWT compared to IDH mutant (IDHMUT) GBM. LAT1 expression, on the other hand, was not different among the individual GBM subtypes. Therefore, we could conclude that FET- and TSPO-PET confer different information on pathological features based on different genetic GBM subtypes and may thus help in planning individualized strategies for brain tumor therapy in the future. A combination of TSPO-PET and FET-PET could be a promising way to visualize tumor-associated myeloid cells and select patients for treatment strategies targeting the myeloid compartment.

The role of miRNAs targeting K-ras and APC genes in colorectal cancer

The occurrence of abnormal expression patterns in different types of cancer suggests that micro RNAs (miRNAs) may play an important role in tumorigenesis. The aim of this study was to examine the expression levels of miRNAs known to be associated with the regulation of the expression levels of the APC and K-ras, which are important in the development of colorectal cancer (CRC). The expression levels of miR-27, miR-663, miR-217, miR-181d, APC and K-ras in the serum, tumor and adjacent tumor-free (healthy) tissues of the patients and serum of the healthy controls were investigated with qRT-PCR. Expression levels of miR-217, mR-181d, miR-663, miR-27 and K-ras were found to be higher in CRC tissues than in adjacent tumor-free tissues of the patients. In patient serum samples, miR-663 levels were statistically more elevated than in controls. In patient tumor tissues, miR-217, miR-181d and miR-27 expressions were found to be higher. Increased miR-181d and miR-217 expression levels are associated with increased K-ras expression in the tumor tissues, and the expression of K-ras, which takes part as an oncogene in the CRC development, might be regulated by these miRNAs (Tab. 4, Ref. 33).

MiRNA-30 Play an Important Role in Non-Small Cell Lung Cancer (NSCLC).

The aim of the study was to explore the expression level of miRNA-30 expression in patients with non-small cell lung cancer (NCLC) and analyze its correlation with clinicopathological features and prognosis. Preoperative serum samples and paracancerous tumor-free tissues of 108 patients with NSCLC treated in our hospital as well as serum samples of 108 healthy subjects were collected from April 2015 to May 2018. The expression levels of miRNA-30 in tissue samples were detected by in situ hybridization and that of miRNA-30 mRNA in serum samples by real-time quantitative PCR (RT-qPCR). The difference in miRNA-30 expression in tumor-free tissues of NSCLC patients and NSCLC tissues of each stage was measured. The miRNA-30 mRNA levels in NSCLC patients was compared with those in healthy subjects. All subjects were divided into low-expression group (< mean) and high expression group (≥ mean) with the mean value of miRNA-30 levels being the critical value. The relationship between miRNA-30 levels and clinicopathological parameters (gender, age, lymph node metastasis, tumor size, TNM stage, degree of infiltration, and differentiation) was analyzed, and the prognosis of different serum miRNA-30 levels was compared based to follow-up data. The expression level of miRNA-30 in cancer tissues and serum of NSCLD patients was significantly lower (p < 0.05, respectively). The results of ROC curve analysis showed that the area under the curve for the diagnosis of NSCLC using miRNA-30 was 0.802 (95% CI: 0.742 to 0.861, p < 0.001), with the diagnostic threshold being 0.798, sensitivity and specificity being 75.9% and 76.0%, respectively. Serum miRNA-30 levels in NSCLC patients were not associated with gender, age, and depth of infiltration (p > 0.05), but correlated with lymph node metastasis, tumor size, TNM stage, and degree of differentiation (p < 0.05). The median overall survival of the miRNA-30 low expression group was 23.0 months, which was shorter than the 36.0 months of high expression group, and the difference was statistically significant (p < 0.05). miRNA-30 is lowly expressed in NSCLC patients and participates in the development of NSCLC. Moreover, NSCLC patients with low expression show poor prognosis. Thus, miRNA-30 features potential as a marker for NSCLC screening and prognosis prediction.

Management of Local Soft Tissue Recurrent Osteosarcoma After Wide Resection and Reconstruction with Megaprosthesis: A Case Report

Introduction: Limb salvage surgery involves all of the surgical methods to achieve the eradication of a malignant neoplasm and restoration of the limb with a satisfactory oncologic, functional, and cosmetic outcome. Rates of local recurrence are 4% to 10%. There is a chance of local recurrence from the contamination of biopsy path. Neoadjuvant chemotherapy has a guarding effect on managing neoplasm infiltration at the biopsy location.Case Presentation: An 18 year old male experienced a new painless lump at his right anterior thigh with size about 5 cm in diameter since 5 months ago. Previously, he underwent limb salvage surgery with megaprosthesis about 6 months ago due to osteosarcoma at his right distal femur. He also got neoadjuvant and adjuvant chemotherapy for 6 cycles. Mass removal was done and 1.5 cm mass in diameter was found within quadriceps muscle with a soft consistency and well-defined border from the previous biopsy site that had not been resected. The specimen result was osteosarcoma surrounded by tumor-free tissue. Postoperatively, he still had the same range of motion function as before.Conclusions: We need to consider the previously contaminated biopsy path that could lead to local recurrence. Factors that affect the prediction of the recurrent disease are the disease-free time period, location of recurrence and histological response to therapy and the capability to achieve total surgical removal. Tumor removal followed by the local radiation and chemotherapy is the preferred treatment for recurrence.

Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer.

Adipose-derived stem cells (ADSCs) have been implicated in tumor growth and metastasis in breast cancer. ADSCs exhibit tumor tropism, and are of increasing clinical relevance due to the autologous fat grafting for breast reconstruction. Although we have previously shown that a high level of the adipocytokine visfatin in human breast cancer tissues correlated with tumor progression mediated by cAbl and STAT3, the effects of visfatin in the tumor microenvironment are unclear. To understand how visfatin modulates breast cancer within the tumor-stromal environment, we examined determinants of breast cancer progression using a visfatin-primed ADSCs-tumor co-culture model. ADSCs were isolated from tumor-free adipose tissue adjacent to breast tumors. ADSCs were treated with or without visfatin for 48 h and then collected for co-culture with breast cancer cell line MDA-MB-231 for 72 h in a transwell system. We found that the MDA-MB-231 cells co-cultured with visfatin-treated ADSCs (vADSCs) had higher levels of cell viability, anchorage independent growth, migration, invasion, and tumorsphere formation than that co-cultured with untreated ADSCs (uADSCs). Growth differentiation factor 15 (GDF15) upregulation was found in the co-culture conditioned medium, with GDF15 neutralizing antibody blocking the promoting effect on MDA-MB-231 in co-culture. In addition, a GDF15-induced AKT pathway was found in MDA-MB-231 and treatment with PI3K/AKT inhibitor also reversed the promoting effect. In an orthotopic xenograft mouse model, MDA-MB-231 co-injected with vADSCs formed a larger tumor mass than with uADSCs. Positive correlations were noted between visfatin, GDF15, and phosphor-AKT expressions in human breast cancer specimens. In conclusion, visfatin activated GDF15-AKT pathway mediated via ADSCs to facilitate breast cancer progression.

Assessment of brain delivery of a model ABCB1/ABCG2 substrate in patients with non-contrast-enhancing brain tumors with positron emission tomography

BackgroundP-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) are two efflux transporters expressed at the blood–brain barrier which effectively restrict the brain distribution of the majority of currently known anticancer drugs. High-grade brain tumors often possess a disrupted blood–brain tumor barrier (BBTB) leading to enhanced accumulation of magnetic resonance imaging contrast agents, and possibly anticancer drugs, as compared to normal brain. In contrast to high-grade brain tumors, considerably less information is available with respect to BBTB integrity in lower grade brain tumors.Materials and methodsWe performed positron emission tomography imaging with the radiolabeled ABCB1 inhibitor [11C]tariquidar, a prototypical ABCB1/ABCG2 substrate, in seven patients with non-contrast -enhancing brain tumors (WHO grades I–III). In addition, ABCB1 and ABCG2 levels were determined in surgically resected tumor tissue of four patients using quantitative targeted absolute proteomics.ResultsBrain distribution of [11C]tariquidar was found to be very low across the whole brain and not significantly different between tumor and tumor-free brain tissue. Only one patient showed a small area of enhanced [11C]tariquidar uptake within the brain tumor. ABCG2/ABCB1 ratios in surgically resected tumor tissue (1.4 ± 0.2) were comparable to previously reported ABCG2/ABCB1 ratios in isolated human micro-vessels (1.3), which suggested that no overexpression of ABCB1 or ABCG2 occurred in the investigated tumors.ConclusionsOur data suggest that the investigated brain tumors had an intact BBTB, which is impermeable to anticancer drugs, which are dual ABCB1/ABCG2 substrates. Therefore, effective drugs for antitumor treatment should have high passive permeability and lack ABCB1/ABCG2 substrate affinity.Trial registrationEuropean Union Drug Regulating Authorities Clinical Trials Database (EUDRACT), 2011-004189-13. Registered on 23 February 2012, https://www.clinicaltrialsregister.eu/ctr-search/search?query=2011-004189-13.

Cancer-Derived Succinate Promotes Macrophage Polarization and Cancer Metastasis via Succinate Receptor.

Macrophages form a major cell population in the tumor microenvironment. They can be activated and polarized into tumor-associated macrophages (TAM) by the tumor-derived soluble molecules to promote tumor progression and metastasis. Here, we used comparative metabolomics coupled with biochemical and animal studies to show that cancer cells release succinate into their microenvironment and activate succinate receptor (SUCNR1) signaling to polarize macrophages into TAM. Furthermore, the results from invitro and invivo studies revealed that succinate promotes not only cancer cell migration and invasion but also cancer metastasis. These effects are mediated by SUCNR1-triggered PI3K-hypoxia-inducible factor 1α (HIF-1α) axis. Compared with healthy subjects and tumor-free lung tissues, serum succinate levels and lung cancer SUCNR1 expression were elevated in lung cancer patients, suggesting an important clinical relevance. Collectively, our findings indicate that the secreted tumor-derived succinate belongs to a novel class of cancer progression factors, controlling TAM polarization and promoting tumorigenic signaling.

Activity of Antioxidant Enzymes in the Tumor and Adjacent Noncancerous Tissues of Non-Small-Cell Lung Cancer.

Lung tissue is directly exposed to high oxygen pressure, as well as increased endogenous and exogenous oxidative stress. Reactive oxygen species (ROS) generated in these conditions play an important role in the initiation and promotion of neoplastic growth. In response to oxidative stress, the antioxidant activity increases and minimizes ROS-induced injury in experimental systems. The aim of the present study was to evaluate the activity of antioxidant enzymes, such as superoxide dismutase (SOD; isoforms: Cu/ZnSOD and MnSOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST), along with the concentration of malondialdehyde (MDA) in tumor and adjacent noncancerous tissues of two histological types of NSCLC, i.e., adenocarcinoma and squamous cell carcinoma, collected from 53 individuals with surgically resectable NSCLC. MDA concentration was similar in tumors compared with adjacent noncancerous tissues. Tumor cells had low MnSOD activity, usually low Cu/ZnSOD activity, and almost always low catalase activity compared with those of the corresponding tumor-free lung tissues. Activities of GSH-related enzymes were significantly higher in tumor tissues, irrespective of the histological type of cancer. This pattern of antioxidant enzymes activity could possibly be the way by which tumor cells protect themselves against increased oxidative stress.