Levels In Tumors Of Patients Research Articles

High‑mobility group box 1 has a prognostic role and contributes to epithelial mesenchymal transition in human hepatocellular carcinoma.

High‑mobility group box 1 (HMGB1), a member of the high‑mobility group protein family, was originally characterized as a non‑histone, nuclear DNA‑binding protein. While the roles of HMGB1 in inflammation and cell differentiation have been previously reported, its role in tumor cell migration and invasion, particularly in hepatocellular carcinoma (HCC), has remained elusive. The present study reported that the expression of HMGB1 in HCC tissues was significantly higher than that in matched tumor‑adjacent tissues (P<0.05). HMGB1 was expressed at significantly elevated levels in tumors of patients with large tumor size, high histological grade and advanced tumor‑node‑metastasis stage (P<0.05). The positive expression of HMGB1 correlated with a poor three‑year overall and disease‑free survival of HCC patients (P<0.05). In addition, HMGB1 was an independent factor for predicting the three‑year overall and disease‑free survival of HCC patients (P<0.05). An in vitro experiment revealed that knockdown of HMGB1 inhibited cell migration and invasion in the HCC cell lines Huh7 and MHCC97H (P<0.05). Furthermore, western blot analysis showed that HMGB1 knockdown markedly inhibited epithelial mesenchymal transition in Huh7 and MHCC97H cells. These results suggested that HMGB1 may be utilized as an independent prognostic marker in HCC and may promote tumor progression by promoting cell migration and invasion.

Evaluation of the ability of adjuvant tamoxifen-benefit gene signatures to predict outcome of hormone-naive estrogen receptor-positive breast cancer patients treated with tamoxifen in the advanced setting

To identify molecular markers indicative of response to tamoxifen and easily implemented in the routine setting, we recently reported three gene signatures that could stratify post-menopausal tamoxifen-treated, estrogen receptor-positive (ER+) patients according to outcome in the adjuvant setting. Here, we evaluated the predictive potential of the total of 14 genes included in the 3 gene signatures using 2 hormone-naïve Dutch ER+ cohorts of a total of 285 recurrent breast cancer patients treated with first-line tamoxifen.mRNA levels were measured by reverse transcriptase quantitative PCR (RT-qPCR) and the length of progression-free survival (PFS) was used as the primary endpoint. A Mann–Whitney U test was used to select for differentially expressed genes between tumors of patients who showed or did not show progressive disease within 6 months after start of tamoxifen treatment. Cox univariate and multivariate regression analysis for PFS were used to further assess their (independent) predictive potential.Five (BCAR3, BCL2, ESR1, IGF1R, and NCOA1) of the 14 genes analyzed showed significantly higher mRNA levels in tumors of patients who showed no disease progression within 6 months. Only BCAR3, BCL2 and NAT1 were significantly associated with a favorable PFS in multivariate analysis that included the traditional predictive factors: age, dominant relapse site, disease-free interval, ER and progesterone receptor (PGR), and adjuvant chemotherapy.This study shows that BCAR3, BCL2 and NAT1 in particular exhibit predictive promise regarding the efficacy of tamoxifen treatment in recurrent disease, in addition to the previously shown favorable outcome in the adjuvant setting.

Abstract 1938: Increased miR-126 and miR-375 expression in primary ER-positive breast cancer predict longer relapse-free time following treatment with tamoxifen.

Abstract Adjuvant tamoxifen treatment provides benefit for millions of women with estrogen receptor positive breast cancer; however, up to 50% of patients experience distant or local recurrence or even die. Tamoxifen resistance is contributed to by host and tumor associated factors and the latter has been particularly attributed to adaptive changes of the tumor involving the regulation of cellular pathways associated with growth, proliferation and apoptosis. Recently, microRNA expression signatures have been appreciated as useful tools to improve the classification of tumors into subtypes and the assessment of tumor heterogeneity. However it is not clear to what extend microRNA signatures derived from the primary tumor can predict treatment outcome. Here we test the hypothesis that distinct microRNAs expressed in the primary tumor can predict the onset of local and distant recurrence following adjuvant tamoxifen. We investigated 115 patients diagnosed with ER positive breast cancer at the Robert Bosch Hospital, Stuttgart, Germany, who had received adjuvant tamoxifen, and had a median follow up of 9.23 years (range: 0.21 - 19.96 years). We compared tumor microRNA expression profiles of 6 patients who developed recurrence under tamoxifen with expression profiles of 6 patients who stayed recurrence-free. Matching of patients was based on age at diagnosis, year of surgery and major histopathological parameters. Candidate microRNAs were identified by global microRNA expression profiling using GeneChip miRNA 2.0 arrays (Affymetrix). Different expression of microRNAs was validated using real-time PCR in 103 tumor samples. We identified miR-126 and miR-375 to be expressed at higher levels in tumors of patients with a lower risk of recurrence (p = 0.0003 and p = 0.0081, respectively). Receiver operator characteristics (ROC) were calculated to further validate their values for predicting breast cancer relapse. The area under the curve (AUC) for miR-126 was 0.6838 (p = 0.0006) and 0.6450 (p = 0.0071) for miR-375, respectively. Kaplan-Meier survival analyses showed that a higher expression of miR-126 and/or miRNA-375 in the patients primary tumor was associated with longer relapse-free time (p = 0.0024 and p = 0.0351, respectively). Our data suggest that miR-126 and miR-375 are promising candidates for further evaluation as tamoxifen outcome predictors in ER-positive breast cancer. R.H. and J.A.K. contributed equally to this work Citation Format: Reiner Hoppe, Joanna Achinger-Kawecka, Stefan Winter, Peter Fritz, Wing-Yee Lo, Werner Schroth, Hiltrud Brauch. Increased miR-126 and miR-375 expression in primary ER-positive breast cancer predict longer relapse-free time following treatment with tamoxifen. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1938. doi:10.1158/1538-7445.AM2013-1938

Generation of a prognostic cancer stem-like gene expression signature in men undergoing radical prostatectomy for localized prostate cancer.

4563 Background: Novel biomarkers are needed to predict recurrence after radical prostatectomy for localized prostate cancer. Recent data suggest that cancer stem-like cells (CSC) are present in prostate tumors, and the CSC phenotype has been associated with an aggressive cancer phenotype. We investigated whether tumor mRNA levels of genes typically expressed by CSC are associated with disease recurrence. Methods: Clinically annotated prostatectomy specimens (FFPE) were used for this nested case-control study. Samples represented men who underwent radical prostatectomy and either experienced a recurrence (PSA or clinical; n=152) or no recurrence (controls; n=124) with minimum 5-year follow-up. Men were excluded if they received neoadjuvant hormone therapy. Cases and controls were frequency matched based on D’Amico risk group. Laser capture microdissection with mRNA extraction and quantitative RT-PCR were used to quantify the expression of 12 candidate CSC-associated genes (ALDH1A1, Axin2, Bmi1, CD133, CD44a, CTNNB1/β-catenin, ITGA2/integrin α2, NANOG, Nkx3-1, Notch1, OCT 4, TACSTD2/Trop2). Univariable and multivariable analyses were conducted to measure associations with recurrence. Results: mRNA data were evaluable for 241 of 276 patients. Univariate Wilcoxon analysis showed that 4 genes (Axin2, p&lt;0.001; CD44a, p=0.036; OCT 4, p=0.017; TACSTD2, p=0.008) were expressed at low levels in tumors of patients whose disease recurred. Recursive partitioning analysis identified 3 genes significantly predictive of disease recurrence (Axin2, NANOG, CTNNB1), with cutpoints yielding odds ratios (OR) of 8.5 (Axin2low/NANOGhigh; n=94, 95%CI 3.7-19.2) and 5.1 (Axin2low/NANOGvery high/CTNNB1low; n=26, 95%CI 1.7-14.8). Conclusions: We identified a novel CSC-associated 3 gene expression signature with the potential to predict recurrence after radical prostatectomy. Axin2 is known to interact with CTNNB1/β-catenin in the Wnt signaling pathway, which in turn has been shown to regulate expression of NANOG, a key protein that mediates pluripotency. In vitro experiments and an independent cohort validation study are planned based on these novel findings.

Noninvasive detection of carboxypeptidase G2 activity in vivo

The pseudomonad protein, carboxypeptidase G2 (CPG2), is a prodrug-activating enzyme utilized in the targeted chemotherapy strategies of antibody- and gene-directed enzyme prodrug therapy (ADEPT and GDEPT). We have developed a noninvasive imaging approach to monitor CPG2 activity in vivo that will facilitate the preclinical and clinical development of CPG2-based ADEPT and GDEPT strategies. Cleavage of the novel reporter probe, 3,5-difluorobenzoyl-L-glutamic acid (3,5-DFBGlu), by CPG2, in human colon adenocarcinoma WiDr xenografts engineered to stably express CPG2, was monitored using (19)F MRSI. The high signal-to-noise ratio afforded by the two MR-equivalent (19)F nuclei of 3,5-DFBGlu, and the 1.4 ppm (19)F chemical shift difference on CPG2-mediated cleavage, enabled the dynamics and quantification of the apparent pharmacokinetics of 3,5-DFBGlu and its CPG2-mediated cleavage in the tumor to be evaluated. In addition, the apparent rate of increase of 3,5-difluorobenzoic acid concentration could also provide a biomarker of CPG2 activity levels in tumors of patients undergoing CPG2-based therapies, as well as a biomarker of treatment response. The addition of in vivo reporter probes, such as 3,5-DFBGlu, to the armamentarium of prodrugs cleaved by CPG2 affords new applications for CPG2 as a gene reporter of transgene expression.

Overexpression of NDRG1: relationship with proliferative activity and invasiveness of breast cancer cell line and breast cancer metastasis

To investigate the relationship between NDRG1 and metastasis of breast cancer and the effects of NDRG1 overexpression on the proliferation and invasion of breast cancer cells. NDRG1 was detected at its protein level by immunohistochemistry (IHC) and its messenger RNA (mRNA) was detected by real-time reverse transcriptase-polymerase chain reaction (real time RT-PCR) in clinical breast cancer specimens. Liposome was used to transiently transfer NDRG1 into MDA-MB-231, a highly invasive human breast cancer cell line. The proliferation of MDA-MB-231 was measured by Bromodeoxy Uridine (BrdU) incorporation assay and the transfection effect on cell cycle distribution was determined by fluorescence assisted cell sorting (FACS). The invasive ability of the transfected cells was investigated by reconstituted matrigel invasion and polycarbonate filters migration experiments. NDRG1 expressions at protein and mRNA levels in tumors of patients with lymph node metastases were significantly lower as compared with those with localized breast cancers (P < 0.01). The amount of NDRG1 mRNA in MCF7, a relatively non-invasive breast cancer cell line, was 10.8 times higher than that in MDA-MB-231 cells (P < 0.01). The BrdU incorporation rate declined significantly (P < 0.05) in NDRG1 overexpressing MDA-MB-231 cells. An increase of the cell population at G(0)/G(1) phase was observed 48 hours post-transfection along with a decrease of cell population at S phase. Overexpression of NDRG1 significantly retarded the invasiveness of MDA-MB-231 cells in matrigel-coated invasion chambers (P < 0.05), when compared to cells transfected with control vectors. However, the migration abilities of cells with or without the transfection were virtually identical. NDRG1 expression reversely correlates with breast cancer metastasis and progression, and may serve as a prognostic biomarker for predicting early metastasis. The inhibition of proliferation and invasion demonstrated by our MDA-MB-231 transfection experiments implies that NDRG1 is a tumor metastasis suppressor gene and may be a new candidate for gene therapy against human breast cancer.

Expression of uridine and thymidine phosphorylase genes in human breast carcinoma.

Uridine phosphorylase (UPase) and an angiogenic enzyme, thymidine phosphorylase (dThdPase) are involved in degradation of the pyrimidine nucleosides through phosphorolysis. The expression levels of UPase and dThdPase are higher in human solid tumors including breast carcinomas than in normal tissues. To clarify the correlation between the expression levels of UPase and dThdPase genes and the clinicopathological factors, mRNA levels of these enzymes were examined by RT-PCR in 43 breast carcinomas. UPase gene expression was not correlated with dThdPase gene expression (regression coefficient R = 0.032). Although the expression level of the dThdPase gene was correlated with angiogenesis, detected by immunostaining endothelial cells (R = 0.66), that of UPase gene was not (R = 0.044). These results suggest that UPase does not have a strong angiogenic activity. The UPase gene expression levels in tumors of patients who relapsed were significantly higher than in those from patients who did not (p = 0.039). Although the expression levels of neither UPase or dThdPase were associated with age, pT, pN, pM, estrogen or progesterone receptor positivity, the patients with the higher levels of UPase gene expression had worse survival (p = 0.0038) than those with lower levels. In contrast, the expression of dThdPase gene was not related to relapse or survival of these patients with breast carcinoma. Our findings suggest that the expression level of UPase gene may be an independent prognostic marker in human breast carcinoma.

Multidrug resistance in ovarian cancer

The development of acquired resistance has limited the effectiveness of chemotherapy in the treatment of ovarian cancer. Experimental model systems were developed to study the mechanisms associated with primary resistance to chemotherapeutic agents and broad cross-resistance (multidrug resistance) which is characteristic of human ovarian cancer. Doxorubicin-resistant cell lines developed in vitro by exposure of a sensitive cell line to increasing concentrations of doxorubicin develop resistance on the basis of a decrease in drug accumulation and have increased expression of the mdr-1 gene. This gene encodes for a membrane glycoprotein and leads to a decreased drug accumulation in drug resistant cell lines. Cell lines established from patients refractory to doxorubicin-containing combinations, however, do not demonstrate a decrease in drug accumulation. Studies are in progress on the measurement of mdr-1 levels in tumors of patients undergoing treatment to determine whether agents, such as verapamil may be useful in the treatment of drug resistant gynecologic cancers. Human ovarian cancer cell lines from drug resistant patients also has been demonstrated to increase levels of glutathione. Lowering of glutathione levels with buthionine sulfoximine (BSO), which irreversibly inhibits the enzyme gamma-glutamyl cysteine synthetase, leads to a marked potentiation of the cytotoxicity of melphalan both in vitro and in vivo in a nude mouse model of human ovarian cancer. Based on those studies, BSO is undergoing toxicologic evaluation before initiation of clinical trials in drug resistant patients. Our studies demonstrate that drug resistance in human ovarian cancer is likely due to interaction of multiple factors. However, biochemical intervention in some of the key steps leading to drug resistance has been demonstrated experimentally feasible and indicates that pharmacologic reversal of drug resistance is a clinical possibility.