Expression Levels Of CCND1 Research Articles

Differential roles of cyclin D1 and D3 in pancreatic ductal adenocarcinoma

BackgroundThe cyclin D1 (CCND1) and cyclin D3 (CCND3) are frequently co-overexpressed in pancreatic ductal adenocarcinoma (PDAC). Here we examine their differential roles in PDAC.ResultsCCND1 and CCND3 expression were selectively suppressed by shRNA in PDAC cell lines with expression levels of equal CCND1 and CCND3 (BxPC3), enhanced CCND1 (HPAC) or enhanced CCND3 (PANC1). Suppression of cell proliferation was greater with CCND3 than CCND1 downregulation. CCND3 suppression led to a reduced level of phosphorylated retinoblastoma protein (Ser795p-Rb/p110) and resulted in decreased levels of cyclin A mRNA and protein. A global gene expression analysis identified deregulated genes in D1- or D3-cyclin siRNA-treated PANC1 cells. The downregulated gene targets in CCND3 suppressed cells were significantly enriched in cell cycle associated processes (p < 0.005). In contrast, focal adhesion/actin cytoskeleton, MAPK and NF B signaling appeared to characterize the target genes and their interacting proteins in CCND1 suppressed PANC1 cells.ConclusionsOur results suggest that CCND3 is the primary driver of the cell cycle, in cooperation with CCND1 that integrates extracellular mitogenic signaling. We also present evidence that CCND1 plays a role in tumor cell migration. The results provide novel insights for common and differential targets of CCND1 and CCND3 overexpression during pancreatic duct cell carcinogenesis.

Elevated Cyclin D1 (CCND1) Expression in the Plasma Cells of Patients with Systemic AL Amyloidosis at Diagnosis Is Associated with Higher Brain Natriuretic Peptide (BNP) Levels, Increased Plasma Cells and Significantly Poorer Survival.

Abstract 2815Poster Board II-791 Background:The clonal plasma cells in AL can be considered malignant because they contain cytogenetic abnormalities, including t(11;14) in 40% to 50% of cases (Blood 2001;98:2266; 2008;111:4700; Haematologica 2009;94:380). By gene expression profiling (GEP) they are also reported to overexpress CCND1 (Blood 2005;105:794). To assess the frequency and significance of differences in CCND1 expression in plasma cells from AL patients at diagnosis, we evaluated the differences in CCND1 expression in CD138+ plasma cells from newly diagnosed patients. We then correlated these differences with features of both the deposition and clonal plasma cell diseases, and with treatments, outcomes and overall survival. Methods:Newly diagnosed AL patients were evaluated and CD138+ cells selected and used for GEP with Affymetrix U133 PLUS2.0 arrays (Affymetrix; Santa Clara, CA) or qRT-PCR as previously described (JNCCN 2007;5:179; Blood 2008;111:549). qRT-PCR was performed using TaqMAN Gene Expression Assays with CCND1 (Hs00277039_m1) and RPLP0 (Hs99999902_m1) primers and probes (Applied Biosystems; Foster City, CA) and 2 myeloma cell lines as positive and negative controls (KMS-12BM, RPMI 8226) on an Mx 3000P platform and related software (Stratagene; La Jolla, CA). The comparative Ct method was used and the amount of target was normalized to the control (2-ΔΔCt) assuming an efficiency of 2. Statistical analyses were performed with GraphPad PRISM (GraphPad; La Jolla, CA). Results:CD138+ cells from 58 newly diagnosed AL patients were evaluated. With GEP (n=16), CCND1 median loge-transformed quantitative expression levels were 11.08 (range, 9.6-11.55) in five versus 4.163 (3.875-5.447) in eleven patients (P < 0.01). With qRT-PCR (n=42), relative CCND1 expression levels were 4.21 (1.76-17.24) in twenty versus 0.014 (0-0.99) in twenty-two patients (P < 0.0001). Overall, 43% (25/58) of patients were CCND1+ and did not differ from CCND1- patients with respect to organ involvement, troponin I, urine total protein, creatinine or alkaline phosphatase levels. The median BNP of CCND1+ patients was 252pg/ml (range 20-1880), significantly higher than that of CCND1- patients (109pg/ml (5-4210), P < 0.05). CCND1+ patients tended to have more plasma cells (13% versus 8%, P = 0.06), higher serum free light chain levels (23.3 versus 14mg/dl, P = 0.12) and more kappa clones (6/25 versus 2/33, P = 0.06), and had fewer intact immunoglobulin M-proteins (4/25 versus 22/33, P < 0.01). There were no differences in frequencies of treatment with stem cell transplant or oral melphalan/dexamethasone or in the rates of complete hematologic responses. CCND1+ patients survived a median of only 27 months compared to 60 months for CCND1- patients (P = 0.02) and had a risk of death of 2.86 (CI 1.18-6.94). Conclusions:Significant differences in CCND1 expression in the plasma cells of AL patients at diagnosis can be appreciated by GEP and qRT-PCR and clearly define two groups, CCND1+ and CCND1-, that differ in baseline BNP levels and features of the clonal plasma cell disease. Despite similar treatments and initial responses, CCND1+ patients have poorer overall survival, possibly due to asymptomatic cardiac involvement at diagnosis and low-level persistence of clonal plasma cells despite treatment, hypotheses that merit prospective evaluation. Disclosures:Comenzo:Millenium: Consultancy, Membership on an entity's Board of Directors or advisory committees.

Quantitative Gene Expression Deregulation in Mantle-Cell Lymphoma: Correlation With Clinical and Biologic Factors

There is evidence for a direct role of quantitative gene expression deregulation in mantle-cell lymphoma (MCL) pathogenesis. Our aim was to investigate gene expression associations with other pathogenic factors and the significance of gene expression in a multivariate survival analysis. Quantitative expression of 20 genes of potential relevance for MCL prognosis and pathogenesis were analyzed using real-time reverse transcriptase polymerase chain reaction and correlated with clinical and genetic factors, tumor morphology, and Ki-67 index in 65 MCL samples. Genomic losses at the loci of TP53, RB1, and P16 were associated with reduced transcript levels of the respective genes, indicating a gene-dosage effect as the pathomechanism. Analysis of gene expression correlations between the candidate genes revealed a separation into two clusters, one dominated by proliferation activators, another by proliferation inhibitors and regulators of apoptosis. Whereas only weak associations were identified between gene expression and clinical parameters or blastoid morphology, several genes were correlated closely with the Ki-67 index, including the short CCND1 variant (positive correlation) and RB1, ATM, P27, and BMI (negative correlation). In multivariate survival analysis, expression levels of MYC, MDM2, EZH2, and CCND1 were the strongest prognostic factors independently of tumor proliferation and clinical factors. These results indicate a pathogenic contribution of several gene transcript levels to the biology and clinical course of MCL. Genes can be differentiated into factors contributing to proliferation deregulation, either by enhancement or loss of inhibition, and proliferation-independent factors potentially contributing to MCL pathogenesis by apoptosis impairment.

Correlation of Quantitative Gene Expression by RQ-PCR with Clinical and Biologic Factors in Mantle Cell Lymphoma.

Mantle cell lymphoma (MCL) is characterized by an aggressive clinical course and the t(11;14) translocation leading to overexpression of cyclin D1 (CCND1). Recently, a relation between the quantitative expression of proliferation associated genes including CCND1 and the survival of MCL patients has been demonstrated using microarray gene expression profiling. In the current study a set of genes (CCND1, CCND3, CDK2, CDK4, p16, p27, Rb, E2F1, MYC, Ki-67, BMI, MCL-1, MDM2, ATM, TP53) involved in cell cycle regulation, related to MCL pathogenesis, or located in critical genomic regions was analyzed by real-time RT-PCR (RQ-PCR) in 74 MCL cases and correlated with clinical and biologic factors. Diagnoses were based on morphology and the detection of the t(11;14) and/or CCND1 overexpression on protein or mRNA level. 62 cases were classical MCL, 12 blastoid variants. To obtain a quantitative estimate of the relation between the expression the CCND1 1.7 kb (short; missing 3′ UTR) and 4.5 kb (long; containing 3′ UTR) mRNA isoform a first amplicon was localized in the CCND1 coding region and a second one in the CCND1 3′ UTR. FISH analysis was performed in a subset of 40 cases for 3q26, 6q27, 9p21, 11q22–q23, 12q13, 13q14, and 17p13. As a marker for proliferation activity the mitotic index (MI) was determined in 51 cases (mitoses/10 high power fields). Survival information was available for all cases, additional clinical information (IPI, B-symptoms, tumor bulk) for 60 cases. In univariable analysis a statistically significant association between gene expression levels and overall survival was identified for MYC (p=0.003), BMI (p=0.027), and CCND1 (coding region; p=0.028). A multivariable analysis including clinical factors of known prognostic impact (IPI score, B-Symptoms, Bulk, blastoid morphology) was carried out for each of the candidate genes individually. In this analysis a significant impact on survival was observed for MYC (p=0.008), BMI (p=0.021), and E2F1 (p=0.042), and a trend for Ki-67 (p=0.054), the ratio CCND1 coding region/CCND1 3′ UTR (p=0.062), and CCND1 coding region (p=0.082). Notably, and in contrast to the other genes of prognostic significance, BMI overexpression was positively correlated with overall survival times. In line with this, there was a negative correlation of BMI levels with proliferation activity as measured by the MI (p<0.001; r=−0.515). Other genes with a negative MI correlation were MCL-1 (p<0.001; r=−0.532) and p27 (p<0.001; r=−0.589), whereas the ratio CCND1 coding region/CCND1 3′ UTR showed a positive correlation (p<0.001; r=0.523). Genomic losses at 9p21 and 17p13 were associated with reduced levels of p16 (p=0.002) and TP53 (p=0.011), respectively, arguing for a pathogenic relevance of these candidate genes in MCL. However, neither p16 nor TP53 expression levels obtained prognostic significance in our MCL series. In conclusion, focused RQ-PCR analysis of individual candidate genes showed a correlation between quantitative gene expression and survival times in MCL. In addition to CCND1 expression levels of MYC, BMI, and E2F1 provided prognostic information also in the context of clinical factors suggesting those factors being valuable survival predictors and important components of MCL pathogenesis.