Radiation induces different changes in expression profiles of normal rectal tissue compared with rectal carcinoma

Prostaglandins support progression of colorectal cancer by several mechanisms. This conclusion is based on epidemiological and drug intervention long-term studies or retrieved from animal and cell culture experiments. The aim of the present study was to map receptor and enzyme expression for prostanoid metabolism in the presence of high or low PGE2 content within colon cancer tissue at primary tumor operation and after short-term preoperative provision of non-steroidal anti-inflammatory drug (NSAID). Twenty-three unselected patients with colon cancer were randomly selected to receive indomethacin (NSAID) or sham treatment for 3 days before surgery. Normal colon and tumor tissue were collected at operation for RNA extraction. Tissue PGE2 levels were measured by radioimmunoassay. Gene expression was quantified by microarray and real-time PCR. COX-1 expression increased proportionally to COX-2 expression in colon cancer tissue from untreated patients. Indomethacin reduced PGE2 content in normal and tumor tissue with subsequently decreased IP, HPGD and PPARgamma receptor expression in both tumor and normal colon tissue, while subtype EP1-4 receptors were not significantly influenced by indomethacin treatment. MPGES-1 expression was not related to overall PGE2 content in tumor and colon tissue, but decreased significantly in normal tissue during indomethacin exposure. Reduction of tumor tissue PGE2 was related to significant alteration in expression of several hundred genes indicating decreased cell cycling and increased apoptosis during indomethacin treatment, probably related to upregulation of acute phase reactants in tumor tissue. Increased prostanoid activity in colon cancer tissue is related to cross-talk between tumor and stroma cells.

Proteolytic Cleavage of the CD44 Adhesion Molecule in Multiple Human Tumors

<p>Supplementary Tables 1-4. Table 1. Prostate cancer risk SNP information and frequency of the risk genotypes in these data (Tumor cohort; n=264) Table 2. The associations of the expression of all genes located within 1 Mb window of each risk SNP in both tumor and normal prostate tissue. Table 3. Significant (False Discovery Rate q<0.1) associations of risk SNPs with the expression of genes in tumor or normal prostate tissue across the genome. Table 4. Top pathways (p<1.0E-03) associated with risk SNPs in tumor and normal tissue.</p>

BackgroundLung cancer is a malignant tumor with a high rate of mortality and metastasis. Recently, extensive research has shown that long non-coding RNAs (lncRNAs) play a crucial role in the development and progression of non-small cell lung cancer (NSCLC). In this paper, we aimed to explore the impact of long intergenic non-coding RNA 00511 (LINC00511) on the development and metastasis of NSCLC.MethodsA dataset containing 501 lung squamous cell carcinoma (LUSC) samples and 49 normal samples was downloaded from The Cancer Genome Atlas (TCGA). The differential gene expression and prognostic potential of LINC00511 in LUSC were analyzed by “limma” in R software. Samples of tumor tissues and normal tissues from 67 patients with NSCLC were obtained, along with clinical features. NSCLC cell proliferation, cell cycle, migration, and invasion were detected by LINC00511 knockdown with Cell Counting Kit-8 (CCK-8), flow cytometry, wound-healing assay, and Transwell experiment. The regulatory relationship between LINC00511 and microRNA (miR)-625-5p, or between miR-625-5p and G1 to S phase transition 1 (GSPT1), was detected by luciferase reporter gene assay. LINC00511, miR-625-5p, and GSPT1 expression in tumor and normal tissues and cells was determined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. A xenograft experiment in nude mice was performed. Ki67 and GSPT1 expression in the tumor tissues of the nude mice was assessed by immunohistochemistry.ResultsLINC00511 expression was clearly higher in the tumor tissues of the NSCLC patients than in normal tissues (P<0.001). High LINC00511 expression was related to larger tumor size, positive lymph node metastasis, advanced TNM stage, and a lower 5-year survival rate. Compared with those of the shNC group, the NSCLC cells of the shLINC00511 group had a prominently lower optical density (OD) 450 value at 72 h, a lower percentage of cells in S phase, a higher relative wound width, and a lower invasive cell number (P<0.01 or P<0.001). LINC00511 promoted GSPT1 expression via suppressing miR-625-5p. Compared with those of the shNC group, the nude mice of the shLINC00511 group had a much lower subcutaneous tumor volume and weight (P<0.05 or P<0.001).ConclusionslncRNA LINC00511 promotes proliferation, invasion, and migration of NSCLC cells by targeting miR-625-5p/GSPT. LINC00511 may be a potential diagnostic marker and therapeutic target for NSCLC.

Previous studies have established the role of phosphorylated form of insulin-like growth factor type 1 receptor (p-IGF1R) as a good candidate for tumor biomarker. The aims of this study were to investigate p-IGF1R expression status in gastric cancer (GC) specimens and to clarify its clinical significance. A total of 78 GC patients treated with radical resection were enrolled in this study. Immunohistochemistry was used to detect p-IGF1R and phosphatase and tensin homolog deleted on chromosome ten (PTEN) protein expression in paired tumor and adjacent normal tissues. Results showed a higher level of p-IGF1R protein expression in tumor tissues than that in normal tissues, and the rate of p-IGF1R protein high/moderate expression in GC and normal tissues was 52.6% (41/78) and 6.4% (5/78), respectively (p < 0.001). In contrast, PTEN protein expression was downregulated in GC, as compared with normal tissues (negative/low expression 49/78 vs. 8/78, p < 0.001). Moreover, PTEN protein downregulation was consistent with p-IGF1R upregulation. Overexpression of p-IGF1R protein was associated with lymph metastasis, clinical stage, and adverse 3-year progression-free survival (PFS). Survival analysis and Cox proportional hazards model revealed that p-IGF1R overexpression was an independent factor in predicting PFS for GC patients, apart from lymph metastasis. In conclusion, p-IGF1R is highly expressed in GC, which may be a novel biomarker to predict the clinical outcome of GC patients.

487 Background: miRNAs have been shown to be involved in tumor initiation, progression and metastasis in many cancers including colorectal cancer. However, miRNA profiling specifically in rectal cancer is not well characterized. Our objective was to generate a miRNA expression profile in locally advanced rectal cancer using formalin fixed paraffin embedded (FFPE) biopsy tissue collected from patients with locally advanced rectal cancer. Methods: We collected pre-treatment biopsy tissue and matched normal tissue from 40 rectal cancer patients treated with pre-operative chemoradiation (CRT) and total mesorectal excision (TME). We extracted 50-1000ng of total RNA from FFPE biopsies and optimized small RNA sample preparation for deep-sequencing. We then performed deep sequencing on biopsy and matched normal tissue and compared miRNA expression in biopsy and normal tissues using paired T-test and multiple testing (Q-bound &lt;0.05) to determine significant miRNA expression changes in rectal cancer. Results: 182 miRNAs were differentially expressed in tumor versus normal tissues (Q-bound &lt;0.05); 15 of these miRNAs showed a greater than 2-fold change in expression in tumor tissue; mir-18a, mir-135b, mir-503, mir-584, mir-106b, mir-224, mir-92a, mir-181d were up-regulated and mir-375, mir-378, mir-378c, mir-137, mir-378, mir-147b, mir-30a were down-regulated. miRNA mir-31 showed the highest up-regulation in tumor tissue (16-fold increase) while mir-215 expression decreased 8-fold in tumor compared to normal tissue. Tumor and normal tissues were completely separated by hierarchical cluster analysis based on their distinctive miRNA profiles. Conclusions: We optimized small RNA sample preparation for deep sequencing of miRNAs, an approach which may be useful for quantifying miRNA expression in tissues with limited starting material. We also identified a novel miRNA expression profile in rectal cancer that may be useful as a diagnostic biomarker of disease.

BackgroundMany studies have documented the abnormal concentrations of major/trace elements in serum or malignant tissues of patients, but very few works systematically tested the concentrations of elements in tumor tissues in comparison with paired adjacent normal tissues from the same patients.MethodsTumor and adjacent normal lung tissues were obtained from 93 patients with previously untreated NSCLC, and 43 patients whose tumor and paired normal lung tissues reached 200 mg or more were selected for measurement of the elements' concentrations using an inductively coupled plasma‐atomic emission spectrometer.ResultsWe found that the concentrations of the 52 elements varied from 0.4 ng/g tissue (Lu, Pd, and Tm) to 1 658 000 ng/g (Na), 1 951 000 ng/g (P), and 2 495 000 ng/g (K). Thirty eight of the 52 (73.1%) elements showed approximately equal concentrations in tumor and adjacent normal lung tissues of the patients. The concentrations of nine elements (K, P, Mg, Zn, Rb, Cu, Se, Cs, and Tl) in tumor samples were significantly higher than their paired normal lung tissues, and five elements (Na, Fe, Cr, Cd, and Ge) exhibited decreased concentrations in cancer samples compared to counterpart normal lung tissues. Low Fe in tumor samples was associated with smoking history, whereas low Cr was associated with histology (squamous cell carcinoma) of the patients.ConclusionsOur results demonstrate that measurement of elements’ concentrations in both cancer and paired normal tissues is important to get insights into the roles of these elements in carcinogenesis, and therapeutic approaches to normalize the elements are warranted to treat NSCLCs.

Esophageal cancer is the third most common cancer in Iran. Neoadjuvant chemoradiotherapy (NCRT) is the appropriate treatment for esophageal cancer. This study investigated the expression of cyclooxygenase (COX)-2 enzyme in normal and tumoral tissues before any treatment in patients with esophageal cancer, this study also assessed the effect of NCRT on the expression of COX-2 enzyme in normal and tumoral tissue in samples derived by surgery furthermore, and this study investigated the relationship between expression of COX-2 enzyme and the pathologic tumor regression grade (PTRG) patients. In this study, a total of 120 patients admitted to Omid Hospital, Imam Reza Hospitals, and Reza-Mashhad Medical Center, who were treated with NCRT, were recruited and the expression of the COX-2 enzyme in normal and tumoral tissues was assessed by immunohistochemistry before and after treatment by an expert pathologist between zero and 300. PTRG was determined by a pathologist after treatment. The mean levels of COX-2 expression, obtained from tumoral and normal tissue baseline biopsy in patients, were 177.69 and 64.29, respectively, while in surgical specimen were 177.25 and 49.84, respectively. A significant association was found between PTRG of surgical specimen and COX-2 expression in normal tissue (baseline biopsy) at diagnosis (P = 0.034). The results indicated that expression of COX-2 in tumoral tissues exceeds the expression of COX-2 in normal tissue of the baseline biopsy. Patients with a high expression of COX-2 in baseline tumor biopsies had less response to treatment of pathology compared to patients with lower expression of COX-2 in baseline tumor biopsies.

Objective To explore the difference of the expression level of FK506 Binding Protein 51 (FKBP51) in colorectal adenocarcinoma and normal colorectal tissues, and the correlation between FKBP51 expression level and clinicopathological characteristics, and to clarify whether FKBP51 is involved in the occurrence and development of colorectal cancer. Methods By immunohistochemical staining [streptavidin-peroxidase (SP) method] and Western blotting methods tested 31 cases of colorectal cancer tumor tissues and normal colorectal tissues far from tumor 5 cm, and explored the expression level difference of FKBP51. Combined with clinical data of patients, results were analyzed by statistical method χ2 test of four case table data. Results The high expression rate of FKBP51 in tumor tissues was 74.19% (23/31 cases), while the high expression rate of FKBP51 in normal tissue was 9.68% (3/31 cases). The difference was significant. The expression level of FKBP51 in patients with colorectal carcinoma had no obvious correlation with gender (P=0.771), age (P=0.474), tumor location (P=0.213), degree of differentiation (P=0.318), lymph node metastasis (P=0.124), distant metastasis (P=0.318) and clinical stage (P=0.171); and the tumor size (P=0.049), depth of invasion related (P=0.031), the difference was statistically significant. Conclusions The expression of FKBP51 in colorectal cancer was strong, while weak expression in normal colorectal tissues. With the increase of tumor infiltration and deepening, the expression of FKBP51 became stronger, which indicated that FKBP51 participated in the genesis and development of colorectal cancer, and it might become a new target for individual therapy of colorectal cancer. Key words: Tacrolimus binding proteins/ME; Colorectal neoplasms/ME

Microsatellite alteration in histologically normal lung tissue of patients with non-small cell lung cancer

BackgroundGene expression profiles of normal and tumor tissue reflect both differences in biological processes taking place in vivo and differences in response to stress during surgery and sample handling. The effect of cold (room temperature) ischemia in the time interval between surgical removal of the specimen and freezing is described in a few studies. However, not much is known about the effect of warm (body temperature) ischemia during surgery.MethodsThree women with primary operable breast cancer underwent in situ biopsies from normal breast and tumor tissue prior to radical mastectomy. Ex vivo biopsies from normal and tumor tissue were collected immediately after surgical excision. The putative effects on gene expression of malignancy (tumor versus normal), surgical manipulation (post- versus pre-surgical) and interaction between the two (differences in effect of surgical manipulation on tumor and normal samples) were investigated simultaneously by Generalized Estimating Equation (GEE) analysis in this self-matched study.ResultsGene set enrichment analysis (GSEA) demonstrates a marked difference in effect of surgical manipulation on tumor compared to normal tissue. Interestingly, a large proportion of pathways affected by ischemia especially in tumor tissue are pathways considered to be specifically up regulated in tumor tissue compared to normal.ConclusionThe results of this study suggest that a large contribution to this differential expression originates from altered response to stress in tumor cells rather than merely representing in vivo differences. It is important to bear this in mind when using gene-expression analysis to deduce biological function, and when collecting material for gene expression profiling.

<title>Abstract</title> Objective(s): 1. The common differential genes of breast cancer and thyroid cancer were identified by Gene Express Omnibus database (GEO). The Cancer Genome Atlas (TCGA) database was used to determine the relationship between the screened differential genes and the clinicopathologic features. Gene set enrichment analysis (GSEA) was used to identify the enrichment pathways of the differential genes in breast and thyroid cancers.2. Retrospective studies were conducted to analyze the relationship between the general characteristics, clinicopathological features, and hormone expression levels in patients with dual cancers of breast cancer combined with thyroid cancer and patients with breast cancer and thyroid cancer alone, and to further analyze the KRT19 expression levels in their tissues. analyze the expression of KRT19 protein in their tissues. Method(s): 1. Download the datasets GSE70947 and GSE3467 from the GEO database, analyze the differential genes of breast cancer, thyroid cancer and normal tissues respectively by using R, and take the intersection of the differential genes of the two tissues to continue the next step of the study. 2. Download the mRNA-seq data of the above differential genes of BRCA and THCA from the TCGA database, and identify the differences in expression of differential genes between normal tissues and tumor tissues by using the above differential gene substitution in R. Gene set enrichment analysis (GSEA) identifies the differences in expression of differential genes enriched in breast and thyroid cancer. Differential gene substitution analysis, and identify the differences in differential gene expression between normal and tumor tissues, Gene set enrichment analysis (GSEA) to identify the pathways of enrichment in breast and thyroid cancers, and then screened out the representative differential gene KRT19 (human cytokeratin 19).3. Collect the mRNA-seq data of BRCA and THCA in the database of TCGA, and use the R language in the substitution analysis of the above differential genes, and identify the differences in differential gene expression between normal and tumor tissues. 3. July 2023 in Zhongshan People's Hospital and puncture or surgical treatment of breast cancer combined with thyroid cancer patient data, a total of 92 cases (experimental group), another randomly collected in the past two years in Zhongshan People's Hospital in the simple breast cancer patients 100 cases (control group 1), thyroid cancer patients, 100 cases (control group 2). 4. the experimental group and the control group 1, 2 of the patient pathology data for Pathological data of patients in experimental and control groups 1 and 2 were retrospectively analyzed, and additional surgical bulk or puncture specimens were subjected to immunohistochemical staining (IHC) to examine the expression of KRT19 protein in the tissues, and to explore whether there was a difference in its expression between experimental and control groups. Result(s): 1. KRT19 mRNA levels were significantly overexpressed in breast and thyroid cancer tissues as analyzed by the GEO database, and KRT19 was associated with clinicopathological features of breast and thyroid cancers as analyzed by the TCGA database, and the GSEA showed that both the breast and thyroid cancer-KRT19 overexpression groups were significantly enriched in the estrogen-responsive pathway.2. In the dual-primary cancer group, of which breast cancer preceded In the dual-primary cancer group, breast cancer preceded thyroid cancer in 86 cases and thyroid cancer preceded in 6 cases, i.e., breast cancer preceded thyroid cancer in the vast majority of patients (&gt; 90%). Compared with the breast cancer group, the dual-primary cancer group was younger at the time of diagnosis, more often in the premenopausal state, with a larger tumor size, and more often positively expressed estrogen receptors (ER) and progesterone receptors (PR), with a statistically significant difference (all <italic>P</italic> &lt; 0.050). Compared with the thyroid cancer group, the odds of tumors occurring bilaterally were increased in the double primary cancer group, and the levels of triiodothyronine (T3) and thyroxine (T4) (both <italic>P</italic> &lt; 0.050) were significantly higher. KRT19 was more frequently positively expressed in breast cancer than in breast cancer alone in dual primary cancers ( <italic>P</italic> = 0.069), and in thyroid cancer than in thyroid cancer alone in dual primary cancers ( <italic>P</italic> &lt; 0.050). Conclusion(s): Pathogenesis correlates between breast and thyroid cancers, estrogen receptor expression is associated with dual carcinogenesis, and KRT19 influences dual carcinogenesis through the estrogen response pathway.

Certain directed oncologic therapies seek to take advantage of the fact that tumors are typically more susceptible to directed therapeutic agents than normal tissue owing to their extensive networks of poorly formed, leaky vasculature. If differences between the vascularity of normal and tumor tissues could be quantified, patients could be selected for or excluded from directed treatments on the basis of this difference. However, angiographic imaging techniques such as digital subtraction angiography (DSA) yield two-dimensional data that may be inadequate for this task. As a first step, the authors evaluated the feasibility of using a commercial implementation of flat panel computed tomography (FPCT) to quantify differences in enhancement of a simulated tumor compared with normal tissue based on differences in CT number measured in precontrast and postcontrast scans. To evaluate the FPCT scanner studied, the authors scanned several phantoms containing simulated normal and tumor tissues. In the first experiment, the authors used an anthropomorphic phantom containing inclusions representing normal, tumor, and bone tissue to evaluate the constancy of CT numbers in scans repeated at clinically relevant intervals of 1 and 3 min. The authors then scanned gelatin phantoms containing dilutions of iodinated contrast to evaluate the accuracy of relative contrast enhancement measurements for a clinical FPCT system. Data were analyzed using widely available software. CT numbers measured in identical locations were constant over both scan intervals evaluated. Measured relative contrast enhancement values were accurate compared with known relative contrast enhancement values. Care must be taken to avoid artifacts in reconstructed images when placing regions of interest. Despite its limitations, FPCT in the interventional laboratory can be used to quantify relative contrast enhancement in phantoms. This is accomplished by measuring CT number in simulated tumor and normal tissue on precontrast and postcontrast scans. This information opens the door for refinement of technique in an effort to use such a technique to plan directed therapies.

25 Background: Long-term prospective data from within the Health Professionals Follow-up Study (HPFS) suggest more frequent ejaculation throughout adult life may be protective against future development of prostate cancer (PCa). Compared to men with an average monthly ejaculation frequency (EF) of 4-7 times/month during adulthood, men who ejaculated at least 21 times per month were 30% less likely to be diagnosed with PCa after adjustment for a number of potential confounders and PCa screening. We sought to explore mechanisms through which EF may influence PCa development utilizing gene expression data assessed on tumor and adjacent normal tissue. Methods: In 1992, 31,925 HPFS participants were questioned on their average monthly EF at ages 20-29, 40-49, and in the previous year; 3,839 of these men were later diagnosed with PCa. Expression of 20,254 genes was available from archival tumor tissue from 156 cases; 84 of these cases also had data from adjacent normal tissue. We regressed each gene expression value as a continuous outcome on EF, age at diagnosis, and year at diagnosis. We calculated the p value associated with the coefficient for the EF variable as a measure of the strength of the trend test. We then used t values for the coefficients to rank the genes, and ran a gene set enrichment analysis to identify pathways with genes associated with EF. Results: We observed that while there was not much signal in the tumor tissue, there was enrichment of smaller p values in the adjacent normal tissue. EF in the time period most proximal to diagnosis is most strongly associated with gene expression in normal tissue. Focusing on the association between gene expression in normal tissue and EF in the year prior to the questionnaire (1991), the top six differentially expressed genes were OR4K1, CCDC138, SERPINA9, TRMT61B, ZNF302, and ASB4. Gene set enrichment analysis revealed a substantial number of pathways differentially expressed at the FDR &lt;0.05 threshold. Conclusions: Identifying changes in gene expression associated with EF in normal prostate tissue provides further support for a biological role of ejaculation frequency in the etiology of PCa. Notably, pathways downregulated with increasing ejaculation frequency were primarily immune related.

Adenosine receptors (A1, A2A, A2B, A3) play an important role in the regulation of growth, proliferation and death of cancer and normal cells. We recently showed the expression profile of A2A and A2B receptors in normal and tumor breast tissues. In the present study, we used semiquantitative RT-PCR to measure the A1 and A3 gene expression levels in normal and tumor breast tissues. Breast tumors (n = 18) and non-neoplastic mammary tissues (n = 10) were collected and histologically confirmed to be neoplastic or non-neoplastic, respectively. Total RNA was extracted and reverse transcribed into cDNA, and PCR was performed under optimized condition for each receptor subtype. Amplification of beta-actin mRNA served as control for RT-PCR. The PCR products were separated on 1.7% agarose gels. The intensity of the bands was quantitated with ImageJ software after normalization against beta-actin expression. All breast tumor and normal tissue specimens expressed A1 and A3 adenosine receptor transcripts. However, we observed that the expression level of the A3 receptor in tumor tissues was 1.27-fold that of normal tissues, whereas there was no significant difference between the expression levels of A1 in normal and tumor tissues. Interestingly, the results of the present study indicate that breast tumors exhibit a higher level of A3 transcripts (than normal tissues) and support the possible key role of A3 adenosine receptor in tumor development. However, further studies based on real-time quantitative RT-PCR are needed to identify the exact gene expression levels.