scholarly journals Μελέτη γλυκοζαμινογλυκανών και πρωτεογλυκανών σε λευχαιμικά κύτταρα

1998 ◽  
Author(s):  
Ευδοκία Μακατσώρη
Keyword(s):  
Western Blotting ◽  
Rt Pcr

Η ΠΑΡΟΥΣΑ ΔΙΑΤΡΙΒΗ ΕΙΧΕ ΣΤΟΧΟ ΤΗ ΜΕΛΕΤΗ ΠΡΩΤΕΟΓΛΥΚΑΝΩΝ (PGS)ΚΑΙ ΓΛΥΚΟΖΑΜΙΝΟΓΛΥΚΑΝΩΝ (GAGS)ΛΕΥΧΑΙΜΙΚΩΝ ΛΕΜΦΟΚΥΤΤΑΡΩΝ ΚΑΙ ΜΟΝΟΚΥΤΤΑΡΩΝ ΚΑΙ ΤΗ ΜΕΛΕΤΗ ΤΗΣ ΔΡΑΣΗΣ ΜΙΤΟΓΟΝΩΝ ΣΤΗ ΣΥΝΘΕΣΗ ΚΑΙ ΚΑΤΑΝΟΜΗ ΤΩΝ GAGS ΟΠΩΣ ΕΠΙΣΗΣ ΤΗ ΜΕΛΕΤΗ ΔΡΑΣΗΣ ΕΞΩΓΕΝΩΝ GAGS ΣΤΟΝ ΚΥΤΤΑΡΙΚΟ ΠΟΛΛΑΠΛΑΣΙΑΣΜΟ,ΜΟΝΩΝ ΤΟΥΣ,Η ΣΕ ΣΥΝΔΥΑΣΜΟ ΜΕ ΜΙΤΟΓΟΝΑ.ΕΠΙΛΕΧΘΗΚΑΝ ΤΡΕΙΣ ΛΕΥΧΑΙΜΙΚΕΣ ΚΥΤΤΑΡΙΚΕΣ ΣΕΙΡΕΣ :JURKAT (ΟΞΕΙΑ Τ-ΛΕΜΦΟΚΥΤΤΑΡΙΚΗ ΛΕΥΧΑΙΜΙΑ),DAUDI (Β-ΛΕΜΦΟΚΥΤΤΑΡΙΚΗ ΛΕΥΧΑΙΜΙΑ)ΚΑΙ THP-1 (ΟΞΕΙΑ ΜΟΝΟΚΥΤΤΑΡΙΚΗ ΛΕΥΧΑΙΜΙΑ).ΑΡΧΙΚΑ ΜΕ RT-PCR ΜΕΛΕΤΗΘΗΚΕ Η ΕΜΦΑΣΗ ΠΡΩΤΕΟΓΛΥΚΑΝΩΝ ΣΕ ΕΠΙΠΕΔΟ MRNA,ΚΑΙ ΒΡΕΘΗΚΕ ΟΤΙ ΚΑΙ ΟΙ ΤΡΕΙΣ ΚΥΤΑΡΙΚΕΣ ΣΕΙΡΕΣ ΕΧΟΥΝ ΤΗ ΔΥΝΑΤΟΤΗΤΑ ΣΥΝΘΕΣΗΣ ΣΥΝΔΕΙΝΗΣ-1,-2,-4,ΓΛΥΠΙΚΑΝΗΣ,CD44,ΒΕΡΣΙΚΑΝΗΣ-0 ΚΑΙ ΒΕΡΣΙΚΑΝΗΣ-1(ΤΥΠΟΣΕΝΑΛΛΑΚΤΙΚΟΥ ΜΑΤΙΣΜΑΤΟΣ ΤΗΣ ΒΕΡΣΙΚΑΝΗΣ). ΤΑ DAUDI ΕΧΟΥΝ ΕΠΙΠΛΕΟΝ ΤΗ ΔΥΝΑΤΟΤΗΤΑ ΣΥΝΘΕΣΗΣ ΙΝΟΜΟΝΤΟΥΛΙΝΗΣ,ΕΝΩ ΤΑ THP-1 ΜΠΟΡΕΙ ΝΑ ΣΥΝΘΕΤΟΥΝ ΠΕΡΛΕΚΑΝΗ ΚΑΙ ΘΡΟΜΒΟΜΟΝΤΟΥΛΙΝΗ. ΠΙΣΤΟΠΟΙΗΘΗΚΕ ΜΕ WESTERN BLOTTING Η ΠΑΡΟΥΣΙΑ ΣΥΝΔΕΙΝΗΣ -1,ΔΥΟΤΥΠΩΝ ΣΥΝΔΕΙΝΗΣ -4 ΔΙΑΦΟΡΕΤΙΚΗΣ ΓΛΥΚΟΖΙΛΙΩΣΗΣ,Η ΜΕΓΑΛΥΤΕΡΗΣ ΘΕΙΩΣΗΣ,ΘΡΟΜΒΟΜΟΝΤΟΥΛΙΝΗΣ, ΒΕΡΣΙΚΑΝΗΣ-0,ΒΕΡΣΙΚΑΝΗΣ-1 ΜΙΑΣ PG ΘΕΙΙΚΗΣ ΚΕΡΑΤΑΝΗΣ ΚΑΙ ΠΕΡΛΕΚΑΝΗΣΑΝΑΛΟΓΑ ΜΕ ΤΗΝ ΚΑΘΕ ΚΥΤΤΑΡΙΚΗ ΣΕΙΡΑ.ΜΕ ΧΡΗΣΗ ΕΝΖΥΜΙΚΩΝ ΚΑΤΕΡΓΑΣΙΩΝ ΚΑΙ ΑΝΑΛΥΣΗΣ HPLC ΒΡΕΘΗΚΕ ΠΩΣ ΚΑΜΙΑ ΚΥΤΤΑΡΙΚΗ ΣΕΙΡΑ ΔΕ ΣΥΝΘΕΤΕΙ HA.ΣΥΝΘΕΤΟΥΝ ΚΥΡΙΩΣ CSΚΑΙ ΣΕ ΜΙΚΡΟΤΕΡΟ ΠΟΣΟΣΤΟ HS.ΑΚΟΛΟΥΘΩΣ,ΕΛΕΓΘΗΚΕ Η ΕΠΙΔΡΑΣΗ ΕΞΩΓΕΝΩΝ GAGS ΣΤΟΝ ΠΟΛΛΑΠΛΑΣΙΑΣΜΟ ΤΩΝ ΚΥΤΤΑΡΩΝ,ΟΠΩΣ ΕΠΙΣΗΣ ΚΑΙ Η ΔΡΑΣΗ ΓΛΥΚΟΖΑΜΙΝΟΓΛΥΚΑΝΩΝ ΠΑΡΟΥΣΙΑ ΜΙΤΟΓΟΤΩΝ ΣΤΟΝ ΠΟΛΛΑΠΛΑΣΙΑΣΜΟ ΤΩΝ ΚΥΤΤΑΡΩΝ ,ΜΕ ΣΚΟΠΟ ΝΑ ΔΙΑΠΙΣΤΩΘΕΙ ΤΟ ΚΑΤΑ ΠΟΣΟ ΣΥΜΜΕΤΕΧΟΥΝ ΟΙ GAGS ΣΤΟΥΣ ΜΗΧΑΝΙΣΜΟΥΣ ΔΙΕΓΕΡΣΗΣ.

scholarly journals The Progesterone Receptor in Human Term Amniochorion and Placenta Is Isoform C

Endocrinology ◽  
2006 ◽  
Vol 147 (2) ◽  
pp. 687-693 ◽  
Author(s):  
Anthony H. Taylor ◽  
Penny C. McParland ◽  
David J. Taylor ◽  
Stephen C. Bell
Keyword(s):  
Progesterone Receptor ◽  
Western Blotting ◽  
Cell Types ◽  
Specific Role ◽  
Fetal Membranes ◽  
Rt Pcr ◽  
Reproductive Tissues ◽  
Pcr Techniques ◽  
Human Parturition ◽  
Extraembryonic Tissues

The mechanism that initiates human parturition has been proposed to be functional progesterone withdrawal whereby the 116-kDa B isoform of the progesterone receptor (PR-B) switches in favor of the 94-kDa A isoform (PR-A) in reproductive tissues. Recently other PR isoforms, PR-S, PR-C, and PR-M generated from the same gene have been identified and partially characterized. Using immunohistochemical, Western blotting, and RT-PCR techniques, evidence is provided that the major PR isoform present in human term fetal membranes (amnion and chorion) and syncytiotrophoblast of the placenta is neither of the classical nuclear PR-B or PR-A isoforms but is the N terminally truncated 60-kDa PR-C isoform. Evidence is also provided that the PR-C isoform resides in the cytoplasm of the expressing cell types. Data are also presented to show that PR-B, PR-A, and PR-S isoforms are essentially absent from the amnion and chorion, whereas PR isoforms A, B, C, and S are all present in the decidua, with PR-A being the major isoform. The syncytiotrophoblast of the placenta contains the cytoplasmic PR-C isoform but not PR-A, PR-B, or PR-S. The major PR isoform in the amnion, chorion, and placenta is PR-C, suggesting that the cytoplasmic PR-C isoform has a specific role in extraembryonic tissues and may be involved in the regulation of human parturition.

scholarly journals Expression of β-catenin in human trophoblast and its role in placenta accreta and placenta previa

2018 ◽  
Vol 47 (1) ◽  
pp. 206-214
Author(s):  
Qing Han ◽  
Lianghui Zheng ◽  
Zhaodong Liu ◽  
Jinying Luo ◽  
Rongxin Chen ◽  
...  
Keyword(s):  
Western Blotting ◽  
Cell Invasion ◽  
Placenta Accreta ◽  
Placenta Previa ◽  
Control Group ◽  
Rt Pcr ◽  
Lesion Area ◽  
Human Trophoblast ◽  
Chorionic Villi ◽  
Pcr Techniques

Objectives To investigate the expression of β-catenin in chorionic villi, and to explore its roles in placenta accreta and placenta previa. Methods We compared β-catenin expression in the control group, placenta accreta group (lesion area and normal zones), and placenta previa group (placental central and placental edge zones) by immunohistochemistry, Western blotting, and RT-PCR techniques. Results Compared with the normal group, the placenta accreta group had a longer length of stay, greater bleeding volume, and lower newborn birth weight. Further, the expression of β-catenin was lower in both placenta previa and placenta accreta groups than in the control group, as measured by immunohistochemistry. Compared with the control group, expression of β-catenin was significantly lower in the placenta previa and placenta accreta groups by Western blotting and RT-PCR. Importantly, the level of placental β-catenin was significantly different when compared between the lesion and normal zones of placenta. Conclusion The expression of β-catenin in placenta accreta might play an important role in the regulation of placental cell invasion; low expression of β-catenin in placenta accreta might be responsible for excessive trophoblastic invasion.

scholarly journals Visfatin Induces Inflammation and Insulin Resistance via the NF-κB and STAT3 Signaling Pathways in Hepatocytes

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Yu Jung Heo ◽  
Sung-E Choi ◽  
Ja Young Jeon ◽  
Seung Jin Han ◽  
Dae Jung Kim ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Signaling Pathways ◽  
Western Blotting ◽  
Insulin Signaling ◽  
Proinflammatory Cytokine ◽  
Hepg2 Cells ◽  
Molecular Mechanisms ◽  
Immunocytochemical Staining ◽  
Mrna Levels ◽  
Rt Pcr

Background. It has been suggested that visfatin, which is an adipocytokine, exhibits proinflammatory properties and is associated with insulin resistance. Insulin resistance and inflammation are the principal pathogeneses of nonalcoholic fatty liver disease (NAFLD), but the relationship, if any, between visfatin and NAFLD remains unclear. Here, we evaluated the effects of visfatin on hepatic inflammation and insulin resistance in HepG2 cells and examined the molecular mechanisms involved. Methods. After treatment with visfatin, the inflammatory cytokines IL-6, TNF-α, and IL-1β were assessed by real-time polymerase chain reaction (RT-PCR) and immunocytochemical staining in HepG2 cells. To investigate the effects of visfatin on insulin resistance, we evaluated insulin-signaling pathways, such as IR, IRS-1, GSK, and AKT using immunoblotting. We assessed the intracellular signaling molecules including STAT3, NF-κB, IKK, p38, JNK, and ERK by western blotting. We treated HepG2 cells with both visfatin and either AG490 (a JAK2 inhibitor) or Bay 7082 (an NF-κB inhibitor); we examined proinflammatory cytokine mRNA levels using RT-PCR and insulin signaling using western blotting. Results. In HepG2 cells, visfatin significantly increased the levels of proinflammatory cytokines, reduced the levels of proteins (e.g., phospho-IR, phospho-IRS-1 (Tyr612), phospho-AKT, and phospho-GSK-3α/β) involved in insulin signaling, and increased IRS-1 S307 phosphorylation compared to controls. Interestingly, visfatin increased the activities of the JAK2/STAT3 and IKK/NF-κB signaling pathways but not those of the JNK, p38, and ERK pathways. Visfatin-induced inflammation and insulin resistance were regulated by JAK2/STAT3 and IKK/NF-κB signaling; together with AG490 or Bay 7082, visfatin significantly reduced mRNA levels of IL-6, TNF-α and IL-1β and rescued insulin signaling. Conclusion. Visfatin induced proinflammatory cytokine production and inhibited insulin signaling via the STAT3 and NF-κB pathways in HepG2 cells.

scholarly journals Regulation and Mechanism of miR-518d through the PPARα-Mediated NF-κB Pathway in the Development of Gestational Diabetes Mellitus

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Hui Qiu ◽  
Xuemin Liu ◽  
Shenshen Yao ◽  
Jiaren Zhou ◽  
Xue Zhang ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Gestational Diabetes ◽  
Gestational Diabetes Mellitus ◽  
Pregnant Women ◽  
Western Blotting ◽  
Transport Process ◽  
Inflammatory Factors ◽  
Rt Pcr ◽  
Expression Levels ◽  
Antagonist Group

Objectives. To observe the role of miR-518d in pregnant women with gestational diabetes mellitus (GDM) and its adjusting effects on PPARα and to explore the regulatory mechanisms of the NF-κB pathway in the development and progression of GDM. Methods. Placenta tissues and peripheral plasma were obtained from pregnant women with normal pregnancy and GDM, respectively, followed by the detections of miR-518d contents by RT-PCR and the expression levels of inflammatory factors using ELISA. Human placenta trophoblast cells (HTR8/SVneo) were cultured under the conditions of physiological glucose (PG group) and high glucose level (HG group). Cells in the HG group were transfected with miR-518d control, mimics, and inhibitors and were separately administered with a PPARα-specific antagonist (GW6471) and PPARα siRNA, and cells were divided into the following groups: HG+miR-518d control group (HGNC group), HG+miR-518d mimic group (HGM group), HG+miR-518d inhibitor group (HGI group), HGI+PPARα antagonist group, and HGI+PPARα siRNA group. The relative expression levels of miR-518d, PPARα, and its downstream genes and NF-κB signalling pathway-related genes were detected by RT-PCR and Western blotting. The contents of inflammatory factors were examined by Western blotting. A dual-luciferase report assay was performed to validate the correlations between miR-518d and PPARα. In this study, mouse GDM models were established to further prove the previous hypothesis with an in vivo experiment. A total of 40 C57BL/6J mice were randomly divided into the following groups: normal diet group (ControlMs), GDM group (GDMMs group), GDM+miR-518d antagomir group, and GDM+miR-518d antagomir+PPARα antagonist group. The mouse model of GDM was established by feeding with combined high-sugar and high-saturated fat diet and injecting streptozotocin (STZ) after 15-day feeding. Female and male mice were cocaged in the number ratio of 2 : 1, and the evidence of vaginal suppository detected in female mice was marked as D0 of pregnancy. The contents of total cholesterol (CH), triglyceride (TG), fast glucose, and insulin (INS) were examined using ELISA, followed by the evaluation of insulin resistance (IR). The related expression levels were also detected with the above methods shown in the previous cell culture. Results. miR-518d has a high expression level in placentas with GDM. As the target gene of miR-518d, PPARα was downregulated with the increased levels of miR-518d. When GDM occurs, inflammatory responses were elevated, stimulating the nuclear transport process of NF-κB. Activated NF-κB triggered the phosphorylation of IKKβ and IκBα. Conclusions. High expression of miR-518d was observed in the development of GDM. In this study, we validated that miR-518d negatively regulates the expression of PPARα and triggers the nuclear transport process of NF-κB and phosphorylation of pathway-associated proteins leading to an inflammatory response and the development of GDM.

scholarly journals LEM-domain proteins are lost during human spermiogenesis but BAF and BAF-L persist

Reproduction ◽  
2017 ◽  
Vol 154 (4) ◽  
pp. 387-401 ◽  
Author(s):  
Razan A Elkhatib ◽  
Marine Paci ◽  
Romain Boissier ◽  
Guy Longepied ◽  
Yasmina Auguste ◽  
...  
Keyword(s):  
Western Blotting ◽  
Nuclear Periphery ◽  
Somatic Cells ◽  
Nuclear Lamina ◽  
Full Length ◽  
Rt Pcr ◽  
Male Pronucleus ◽  
Spermatid Nucleus ◽  
Chromatin Proteins

During spermiogenesis the spermatid nucleus is elongated, and dramatically reduced in size with protamines replacing histones to produce a highly compacted chromatin. After fertilisation, this process is reversed in the oocyte to form the male pronucleus. Emerging evidence, including the coordinated loss of the nuclear lamina (NL) and the histones, supports the involvement of the NL in spermatid nuclear remodelling, but how the NL links to the chromatin is not known. In somatic cells, interactions between the NL and the chromatin have been demonstrated: LEM-domain proteins and LBR interact with the NL and respectively, the chromatin proteins BAF and HP1. We therefore sought to characterise the lamina-chromatin interface during spermiogenesis, by investigating the localisation of six LEM-domain proteins, two BAF proteins and LBR, in human spermatids and spermatozoa. Using RT-PCR, IF and western blotting, we show that six of the proteins tested are present in spermatids: LEMD1, LEMD2 (a short isoform), ANKLE2, LAP2β, BAF and BAF-L, and three absent: Emerin, LBR and LEMD3. The full-length LEMD2 isoform, required for nuclear integrity in somatic cells, is absent. In spermatids, no protein localised to the nuclear periphery, but five were nucleoplasmic, receding towards the posterior nuclear pole as spermatids matured. Our study therefore establishes that the lamina-chromatin interface in human spermatids is radically distinct from that defined in somatic cells. In ejaculated spermatozoa, we detected only BAF and BAF-L, suggesting that they might contribute to the shaping of the spermatozoon nucleus and, after fertilisation, its transition to the male pronucleus.

High Expression of Human Amelogenin in E. Coli

1996 ◽  
Vol 10 (2) ◽  
pp. 187-194 ◽  
Author(s):  
D. Deutsch ◽  
E. Chityat ◽  
M. Hekmati ◽  
A. Palmon ◽  
Y. Farkash ◽  
...  
Keyword(s):  
Amino Acid ◽  
Western Blotting ◽  
Rt Pcr ◽  
Amino Acid Sequencing ◽  
Terminal Amino Acid ◽  
Expression Plasmid ◽  
Over Expression ◽  
E Coli ◽  
Sds Page ◽  
Terminal Amino

A human cDNA, encoding for the 175-aminoacid human amelogenin, was prepared by RT PCR from tooth bud mRNA and sub-cloned into pGEX-KG expression plasmid for over-expression in E. coli. The expressed protein was characterized by SDS-PAGE, Western blotting, and N-terminal amino acid sequencing.

scholarly journals Taurocholate Induces Connective Tissue Growth Factor Expression in Hepatocytes Through ERK-YAP Signaling

2018 ◽  
Vol 50 (5) ◽  
pp. 1711-1725
Author(s):  
Bin Yu ◽  
Guan-nan Jin ◽  
Mei Ma ◽  
Hui-fang Liang ◽  
Bi-xiang Zhang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Liver Fibrosis ◽  
Western Blotting ◽  
Tissue Growth ◽  
Hepatic Cell ◽  
Erk Signaling ◽  
Rt Pcr ◽  
Hepatic Cell Lines

Background/Aims: Cholestasis is characterized by intrahepatic accumulation of cytotoxic bile acids (BAs), ultimately leading to fibrosis and cirrhosis, but the precise role of BAs in cholestasis-induced liver fibrosis remains largely elusive. In this study, we investigated the role and the potential mechanisms of BAs during cholestasis in vivo and in vitro. Methods: The effect of BAs during cholestasis was studied in bile duct ligation (BDL) rat models in vivo. We performed immunohistochemistry, Western blotting, and quantitative RT-PCR to investigate the expression of connective tissue growth factor (CTGF/CCN2) in rat liver during cholestasis. The hepatic cell lines AML12 and BRL were stimulated with taurocholate (TC) and the level of CTGF/CCN2, and activation of ERK, Akt, p38 MAPK, JNK, YAP, and TGF-β/Smad signaling were examined using Western blotting. Next, to elucidate the mechanism underlying bile acid-induced CTGF/CCN2, we treated the cells with MEK1/2 inhibitor (U0126), YAP function inhibitor (verteporfin), p38 kinase inhibitor (SB203580), Akt inhibitor (MK2206), and small interfering RNA (siRNA) targeting mek1, erk, and yap in cooperation with TC. Besides, we confirmed the activation of these signaling pathways in BDL and sham rat livers by immunohistochemistry, Western blotting, and quantitative RT-PCR. Results: In this study, we confirmed that the expression of CTGF/CCN2 was increased in BDL-induced rodent cholestatic liver fibrosis. In addition, we showed that TC, the main component of BAs, enhanced the synthesis of CTGF/ CCN2 in AML12 and BRL hepatic cell lines. Moreover, we demonstrated that TC activated ERK, Akt, and YAP signaling in hepatocytes, but the precise roles of these signaling cascades in the expression of CTGF/CCN2 were different: TC-induced expression of CTGF/CCN2 was mediated by ERK-YAP signaling, whereas Akt signaling inhibited ERK signaling and YAP and subsequently the expression of CTGF/CCN2 in hepatocytes. Furthermore, YAP functioned as a downstream regulator of ERK signaling in TC-induced CTGF/CCN2 expression in hepatocytes. Conclusion: Our report provides evidence for the role of conjugated BAs in liver fibrosis and suggests that the production of CTGF/CCN2, induced by conjugated BAs via ERK-YAP axis activation, may be a therapeutic target in cholestasis-induced liver fibrosis.

Nachweis der Osteokalzinexpression osteoblastärer Zellen mandibulären Ursprungs, wachsend auf Biomaterialien, mittels RT-PCR und SDS-PAGE/Western Blotting

2003 ◽  
Vol 7 (5) ◽  
pp. 294-300 ◽  
Author(s):  
D. Turhani ◽  
C. Item ◽  
D. Thurnher ◽  
D. Kapral ◽  
B. Cvikl ◽  
...  
Keyword(s):  
Western Blotting ◽  
Rt Pcr ◽  
Sds Page

Clonal and Subclonal Cytogenetic Aberrations: Association with D-Type Cyclin Expression and Event-Free Survival (EFS) in Multiple Myeloma (MM).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3439-3439
Author(s):  
Dirk Hose ◽  
John DeVos ◽  
Nadine Müller ◽  
Jean-Francois Rossi ◽  
Christiane Heiß ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Western Blotting ◽  
Plasma Cells ◽  
Early Event ◽  
Expression Data ◽  
Rt Pcr ◽  
Qrt Pcr ◽  
Cytogenetic Aberrations ◽  
Significant Difference

Abstract AIM. Expression changes of D-type cyclins are thought to be an early event in the genesis of Multiple Myeloma and are associated with distinct cytogenetic aberrations. These aberrations appear with different percentages (“clonal” or “subclonal”) in a given patient. We assessed whether the height of CCND expression assessed by gene expression profiling and quantitative RT-PCR (qRT-PCR) correlates with the presence of clonal or subclonal aberrations of 11q13, t(11;14) and t(4;14). PATIENTS AND METHODS. 128 newly diagnosed MM-patients (65 training (TG)/63 independent validation group (VG)) and 14 normal donors (ND) were included. Bone marrow aspirates were CD138-purified by activated magnetic cell sorting. RNA was in-vitro transcribed and hybridised to Affymetrix HG U133 A+B GeneChip (TG) and HG U133 2.0 plus array (VG). CCND1 and CCND2 expression was verified by real time RT-PCR and western blotting. iFISH was performed on purified MM-cells using probesets for chromosomes 1q21, 9q34, 11q23, 11q13, 13q14, 15q22, 17p13, 19q13, 22q11 and the translocations t(4;14) and t(11;14). Clonal aberrations were defined as being present in >60%, subclonal aberrations in 20 to 60% of MMC in a given patient. Expression data were gcrma normalised and a Kruskal-Wallis rank sum test used (Bioconductor). RESULTS. 11q13+. CCND1 (208711_s_at, 208712_at) is significantly higher (p<0.0001), CCND2 (200953_s_at, 200951_s_at) significantly lower (p<0.0001) expressed in MMC harbouring clonal, compared to subclonal, or no gain of 11q13. t(11;14). CCND1 is significantly higher (p<0.0001), CCND2 significantly lower (p<0.0001) expressed in MMC harbouring clonal, compared to subclonal, or no t(11;14). t(4;14). CCND1 is significantly lower (p<0.0001), CCND2 significantly higher (p<0.0001) expressed in MMC harbouring clonal compared to subclonal, or no t(4;14). The expression of CCND3 (201700_at) is not significantly different between the 3 groups for all aberrations investigated. CCND2 and CCND3, but not CCND1 are expressed by normal plasma cells. Results have been verified by qRT-PCR (n=40) for CCND1 and CCND2. Expression of CCND1, CCND2 and CCND3 has been verified by western blotting on selected samples. The expression of CCND2 correlates with short EFS, but not if patients with t(4;14) are excluded. There is no significant difference in EFS for patients harbouring the respective aberrations in a clonal or subclonal pattern. CONCLUSION. An additional copy of 11q13 or t(11;14) correlates with increased CCND1 and decreased CCND2 expression, a t(4;14) is associated with an increase of CCND2 and a decrease of CCND1 expression. In each case, the height of the CCND-expression is significantly different whether the respective aberration is clonal or subclonal. Thus, when interpreting expression data in the context of cytogenetic aberrations, it is important to consider if plasma cells carry a respective aberration in a subclonal/clonal pattern.

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