Cloning of a gene highly overexpressed in cancer coding for a novel KH-domain containing protein

Müeller-Pillasch, Friederike; Lacher, Ulrike; Wallrapp, Christine; Micha, Anne E.; Zimmerhackl, Frank; Hameister, Horst; Varga, Gábor; Friess, Helmut; Büchler, Markus W.; Beger, Hans G.; Vilà, Maya R.; Adler, Guido; Gress, Thomas M.

doi:10.1038/sj.onc.1201110

Cited by 259 publications

(264 citation statements)

References 8 publications

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“…The recombinant protein expressed in E. coli BL21 (DE3) was purified using nickel column chromatography. Koc cDNA cloned in the pcDNA3 vector [25] was similarly subcloned to pET28a vector and recombinant protein expressed as above. Imp1 construct pCMV5-Imp1 was kindly provided by F.C.…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

“…Nielsen [26]. Anti-Koc/Imp3 was raised against specific C-terminal peptides of the respective proteins [25] and anti-p62 raised against the fulllength protein [5]. Reactivities of p53, c-myc, cyclin B1 and p16 were determined with monoclonal antibodies obtained from Oncogene Research Products, Boston, MA.…”

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

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Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocellular carcinoma

Zhang

Megliorino

Peng

et al. 2007

Journal of Hepatology

113

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Background/Aims-Previous studies have demonstrated that during transition from chronic liver disease to hepatocellular carcinoma (HCC), autoantibodies can appear which are not detected in the prior pre-malignant conditions. These antibody responses may be stimulated by cellular proteins involved in carcinogenesis. This study determines the prevalence of antibodies to a selected panel of eight tumor-associated antigens (TAAs) in sera from patients with chronic hepatitis, liver cirrhosis and HCC, and considers the possibility and usefulness of antibodies to such a panel of TAAs in differentiating between these conditions. The panel of eight TAAs includes Imp1, p62, Koc, p53, cmyc, cyclin B1, survivin and p16 full-length recombinant proteins.Methods-Enzyme-linked immunosorbent assay (ELISA) was used to detect antibodies against eight selected TAAs in 30 sera from chronic hepatitis, 30 from liver cirrhosis, and 142 from HCC. Positive results were also confirmed by slot blot, Western blotting and immunoprecipitation assay.Results-Antibody frequency to any individual TAA in HCC varied from 9.9%-21.8%. With the successive addition of TAAs to a final total of eight antigens, there was a stepwise increase of positive antibody reactions reaching a frequency of 59.8% with whole cohort of HCC patients. This was significantly higher than the frequency of antibodies in chronic hepatitis (20%), liver cirrhosis (30%) and normal individuals (12.2%).Conclusions-This study demonstrates that malignant transition to HCC is associated with increased autoantibody responses to certain cellular proteins which might have a role in tumorigenesis, and shows that a mini-array of eight TAAs enhanced antibody detection for diagnosis of HCC. More studies in patients with HCC and precursor conditions such as chronic hepatitis, alcoholic hepatitis and liver cirrhosis using enlarged TAA mini-array panels might further improve the sensitivity and specificity of this mode of cancer immunodiagnosis. Its additional usefulness might be in the early detection of cancer in some patients with predisposing conditions.

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Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

Section: Expression and Purification Of Recombinant Proteinsmentioning

confidence: 99%

Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocellular carcinoma

Zhang

Megliorino

Peng

et al. 2007

Journal of Hepatology

113

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“…Originally, IGF2BP3 was found to be overexpressed in pancreatic cancer. 11 More recently, increased IGF2BP3 protein expression has been reported in a variety of solid tumors and was found to be associated with a higher histological grade, 12 progression or metastasis 13,14 and with an unfavorable outcome. [15][16][17][18] These data indicate that IGF2BP3 may promote tumor cell proliferation and progression by enhancing signaling via the IGF pathway.…”

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confidence: 99%

Increased tumor cell proliferation in mantle cell lymphoma is associated with elevated insulin-like growth factor 2 mRNA-binding protein 3 expression

et al. 2012

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Mantle cell lymphoma is an aggressive, non-curable B-cell lymphoma, characterized by the translocation t(11;14)(q13;q32) involving CCND1 and a high number of additional genetic alterations. Chromosomal gains of 7p are frequent in mantle cell lymphoma, with insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3 aka IMP3) being the most upregulated gene in this region. IGF2BP3 is a member of the IGF II mRNA-BP family, and increased IGF2BP3 expression is associated with an aggressive behavior in many malignant tumors. We here analyze selected genes related to IGF signaling in gene expression and genomic array data of 8 mantle cell lymphoma cell lines and 12 primary mantle cell lymphomas and study IGF2BP3 protein expression in 172 wellcharacterized primary mantle cell lymphomas by immunohistochemistry. The majority of mantle cell lymphoma cell lines and primary cases showed elevated IGF2BP3 mRNA expression and a subset also expressed the IGF1 and IGF2 receptors. On the protein level, 66 of 172 primary mantle cell lymphomas showed IGF2BP3 expression in 450% of tumor cells, and strong IGF2BP3 protein expression was highly associated with increased proliferation as measured by the Ki-67 index, but not with overall survival of mantle cell lymphoma patients. Only a subset of mantle cell lymphomas with marked IGF2BP3 expression had an underlying chromosomal gain in 7p, suggesting that additional mechanisms are involved in the upregulation of IGF2BP3 in mantle cell lymphoma. In seven paired mantle cell lymphoma samples, IGF2BP3 protein expression remained constant between primary diagnosis and relapse. Increased IGF2BP3 expression and, potentially, enhanced IGF signaling may contribute proproliferative stimuli in the evolution of mantle cell lymphoma tumor cells.

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“…However, the precise role of KOC in tumour biology remains to be elucidated. Our preliminary data suggested that KOC is exclusively expressed in tumours and embryonic tissues (Mueller-Pillasch et al, 1997, 1999 and may therefore represent an ideal target for novel diagnostic approaches. To test this hypothesis, a prospective study was undertaken to determine the diagnostic accuracy of measuring KOC expression as compared to the cytological assessment in a consecutive series of fine-needle aspirates (FNAs) from abdominal lesions, aszites, various cysts and cerebrospinal fluid.…”

mentioning

confidence: 81%

“…The KOC assay could be useful to facilitate screening for malignant disease and to improve the diagnostic accuracy of FNAs. In a large-scale screen for differentially expressed genes in pancreatic cancer, we have recently identified a gene encoding a novel protein with four K-homologous (KH) domains, which is highly overexpressed in pancreatic cancer (Mueller-Pillasch et al, 1997). The new gene was named KOC (KH-domain containing protein overexpressed in cancer).…”

mentioning

confidence: 99%

KOC is a novel molecular indicator of malignancy

et al. 2003

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The detection of malignant cells in fine-needle aspirates (FNA's) using marker genes is hampered by the fact that these markers are only expressed by certain malignancies or lack sensitivity and/or specificity. Here we report the results of a prospective pilot study examining the expression of KOC (KH-domain containing protein over expressed in cancer), a novel onco-foetal gene, in 76 patients who underwent fine-needle aspiration for further diagnosis of abdominal lesions, aszites, cysts or cerebrospinal fluid. Aspirates were examined by cytology and by a KOC RT -PCR assay. KOC expression was a highly sensitive and specific indicator of malignancy. The KOC assay could be useful to facilitate screening for malignant disease and to improve the diagnostic accuracy of FNAs. In a large-scale screen for differentially expressed genes in pancreatic cancer, we have recently identified a gene encoding a novel protein with four K-homologous (KH) domains, which is highly overexpressed in pancreatic cancer (Mueller-Pillasch et al, 1997). The new gene was named KOC (KH-domain containing protein overexpressed in cancer). Two known functions of KHdomain containing proteins are the regulation of mRNA stability and subcellular localisation, both of which are implicated in fundamental biological processes such as development, cell growth, differentiation and carcinogenesis (Ross et al, 1997). Therefore, KOC may play a role in the regulation of tumour cell proliferation by interfering with transcriptional and/or posttranscriptional processes. However, the precise role of KOC in tumour biology remains to be elucidated. Our preliminary data suggested that KOC is exclusively expressed in tumours and embryonic tissues (Mueller-Pillasch et al, 1997, 1999 and may therefore represent an ideal target for novel diagnostic approaches. To test this hypothesis, a prospective study was undertaken to determine the diagnostic accuracy of measuring KOC expression as compared to the cytological assessment in a consecutive series of fine-needle aspirates (FNAs) from abdominal lesions, aszites, various cysts and cerebrospinal fluid. MATERIALS AND METHODS Determination of KOC expression in cell lines and FNAsHuman Panc-1 pancreatic cancer cells were purchased from the American Type Culture Collection. Stocks were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v v À1 ) foetal bovine serum (FBS) in a humidified atmosphere of 5% CO 2 /95% air at 371C and passaged every 3 days. Confluent cultures of Panc-1 cells were trypsinised and resuspended in serum-free DMEM. Various numbers of Panc-1 cells were subsequently mixed with 1 ml of human serum and then immediately centrifuged at 1200 r.p.m. for 2 min. The supernatants were decanted. The remaining cell pellets were resuspended in the RNeasy Mini Kit lysis buffer (Qiagen, Hilden, Germany) and total RNA was extracted using the same kit according to the manufacturer's instructions. cDNA synthesis and PCR were performed with the one-step RT -PCR (Polymerase Chain Reaction) Kit ...

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Cloning of a gene highly overexpressed in cancer coding for a novel KH-domain containing protein

Cited by 259 publications

References 8 publications

Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocellular carcinoma

Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocellular carcinoma

Increased tumor cell proliferation in mantle cell lymphoma is associated with elevated insulin-like growth factor 2 mRNA-binding protein 3 expression

KOC is a novel molecular indicator of malignancy

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