bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2088: miR-133a as a tumor suppressive microRNA by targeting multiple oncogenes in head and neck squamous cell carcinoma (HNSCC)
Authors: Hanazawa Toyoyuki , Kikkawa Naoko , Nohata Nijoro , Suzuki Homare , Chiyomaru Takeshi , Enokida Hideki , Nakagawa Masayuki , Okamoto Yoshitaka , Seki Naohiko 
Published: February 2011

Abstract
Head and neck squamous cell carcinomas (HNSCC) represent 6% of all cancers. Despite of considerable advances in surgery, radiotherapy and chemotherapy, the overall 5-year survival rate for patients is less than 40% in advanced stage. The understanding of the new molecular pathways of HNSCC carcinogenesis would be helpful in improving diagnosis and therapy of the disease. MicroRNA(miRNA) are endogenous short non-coding RNA molecules, which regulate gene expression by repressing translation or cleaving RNA transcripts in a sequence-specific manner. miRNA over-expression could result in down-regulation of tumor suppressor genes, whereas their under-expression could lead to oncogenes activation. Expression levels of 665 human mature miRNAs were screened using the stem-loop real-time quantitative PCR method. We identified several down-regulated miRNAs based on miRNA expression signatures in HNSCC. A comparison of miRNA signatures from HSCC and our previous reports (Ichimi et al., Int J Cancer 2009, 125:342-352; Kano et al., AACR 2009, #552; Kikkawa et al., AACR 2009, #587) identified miR-133a that is down-regulated in common, suggesting that miR-133a is candidate tumor suppressor. Gain-of-function analysis revealed that miR-133a transfectant inhibits cell proliferation and cell invasion in cancer cells. For target genes silenced by miR-133a in cancer cells, we performed genome-wide screening (oligo microarray analysis and public database searche). Among the genome-wide analysis, several candidate genes were identified, such as, fascin homolog 1 (FSCN1) and glutathione S-transferase P1 (GSTP1). We focused on FSCN1, an actin-bundling protein, which induces parallel actin bundles in cell protrusions and increases cell motility in several human cancers. The miR-133a which has conserved sequence in the 3′UTR of FSCN1, inhibited of FSCN1 expression. The signal from a luciferase reporter assay was significantly decreased at miR-133a site, suggesting miR-133a directly regulate FSCN1. An FSCN1 loss-of-function assay found significant cell growth and invasion inhibition in HNSCC cells implying an FSCN1 is associated with HSCC carcinogenesis. Tumor suppressive miRNAs and target oncogenes may provide new insights into the mechanisms in cancer. Our findings have therapeutic implications and may be exploited for future HSCC treatments.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2088.
Abstract 2090: MicroRNAs 221/222 and genistein mediated regulation of ARHI tumor suppressor gene in prostate cancer
Authors: Chen Yi , Deng Guoren , Majid Shahana , Saini Shranjot , Zaman Saif , Khatri Gaurav , Suh Seong O. , Liu Jan , Tanaka Yuichiro , Dahiya Rajvir 
Published: February 2011

Abstract
ARHI, an imprinted tumor suppressor gene, is expressed in normal immortalized prostate epithelial cells, but is dramatically down-regulated in prostate cancer cell lines. Here we investigated the mechanisms of ARHI silencing in prostate cancer through microRNA and genistein (a dietary isoflavone from soy) mediated pathways. First, we evaluated ARHI mRNA levels by real time PCR using Q-PCR slice (48 prostate tumor and normal adjacent tissues) and protein levels by immunostaining of prostate tissue array (36 cancer, 8 matched normal adjacent tissue and 4 bone metastases). Both experiments showed down-regulation of ARHI in prostate cancer tissues compared to adjacent normal tissues. To understand the functional significance of ARHI down-regulation with regard to tumorigenesis, ARHI was over-expressed in prostate cancer PC-3 cells by introduction of ARHI plasmid followed by functional studies. Over-expression of ARHI can inhibit cell proliferation, colony formation, invasion and induced caspase-independent apoptosis. Further studies on new mechanism of ARHI down regulation showed a significant inverse relationship between ARHI and miR-221 & 222 which were up-regulated in cancer cell lines. Transfection of miR-221 & 222 inhibitors into PC-3 cells caused a significant induction of ARHI expression. A direct interaction of miR-221 or 222 with a target site on the 3′UTR of ARHI was confirmed by a dual luciferase pMIR-REPORT assay. Finally, we also found that genistein up-regulated ARHI by down regulating miR-221 & 222 in PC-3 cells. In conclusion, ARHI is a tumor suppressor gene down regulated in prostate cancer, and over-expression of ARHI can inhibit cell proliferation, colony formation and invasion. This study demonstrates for the first time that prostate cancer cells have elevated levels of miR-221 & 222 which bind to the 3′UTR of ARHI contributing to its down regulation. Genistein, a potential non-toxic chemopreventive agent restores expression of ARHI and may be an important dietary therapeutic agent for treating prostate cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2090.
Abstract 2092: Control of histone 3 lysine 27 methylation by microRNA 520b in medulloblastoma
Authors: Alimova Irina , Venkataraman Sujatha , Patel Purvi , Harris Peter , Vibhakar Rajeev 
Published: February 2011

Abstract
MicroRNAs are small non-coding RNAs with gene expression regulation at posttranscriptional and transcriptional level. MicroRNAs are widely known to play a key role in carcinogenesis. We investigated the role of microRNAs in medulloblastoma, the most common malignant brain tumor of childhood. We found that microRNA 520b is over expressed in medulloblastoma compared to normal cerebellum. We next examined the biological significance of microRNA 520b. Transfection of medulloblastoma cells (DAOY and ONS76) with microRNA 520b significantly increased cell proliferation and enhanced anchorage-independent growth in soft agar compared to parental cells. Conversely, over expression of anti-microRNA 520b in medulloblastoma cells decreases cell proliferation and colony formation in soft agar. Bio-informatics analysis revealed several chromatin-remodeling proteins as potential targets of microRNA 520b. One of these, SUV39H1 is a histone methyl transferase that methylates histone 3 lysine 27 and suppresses activation of associated promoters. It is known that histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development. We found that inhibition of microRNA 520b results in increased expression of SUV39H1 and subsequent methylation of Histone 3 Lys27 in medulloblastoma cells. SUV39H1 is known to be a tumor suppressor and is mutated in a subset of medulloblastoma. These data indicate that microRNA 520b can function as an oncogene in medulloblastoma by regulating histone modifications.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2092.
Abstract 2097: microRNA profile and identification of miR-29 as a prognostic marker and pathogenetic factor by targeting CDK6 in mantle cell lymphoma
Authors: Zhao Jian-jun , Lin Jianhong , Lwin Tint , Yang Hua , Guo Jianping , Kong William , Dessureault Sophie , Moscinski Lynn , Rezania Dorna , Dalton William S. , Sotomayor Eduardo , Tao Jianguo , Cheng Jin Q. 
Published: February 2011

Abstract
ABSTRACT: Mantle cell lymphoma (MCL) is one of the most aggressive B-cell lymphomas. While a number of protein-coding genes are altered, expression signature and importance of microRNA (miRNA) have not been well documented in this malignancy. Here, we performed miRNA expression profile in 30 patients with MCL using a platform containing 515 human miRNAs. Eighteen miRNAs were downregulated and 21 were upregulated in MCL when compared with normal B lymphocytes. The most frequently altered miRNAs are decrease of miR-29a/b/c, miR-142-3p/5p and miR-150 and increase of miR-124a and miR-155. Notably, expression levels of miR-29 family are associated with prognosis. The patients with significant downregulated miR-29 had short survival as compared to those who express relative high level of miR-29. Prognostic value of miR-29 is comparable with MIPI (MCL International Prognostic Index). Furthermore, we demonstrate miR-29 inhibition of CDK6 protein and mRNA levels by direct binding to 3′UTR. Inverse correlation between miR-29 and CDK6 was observed in MCL. Since cyclin D1 overexpression is a primary event and exerts its function through activation of CDK4/6, our results in primary MCL cells indicates that downregulation of miR-29 could cooperate with cyclin D1 in MCL pathogenesis. Thus, our findings provide not only miRNA expression signature but also a novel prognostic marker and pathogenetic factor for this malignancy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2097.
Abstract 2098: The receptor tyrosine kinase AXL is a target for the human miR-34a microRNA
Authors: Mackiewicz Mark , Huppi Konrad , Cornelison Robert , Caplen Natasha J. 
Published: February 2011

Abstract
Determining the protein targets of microRNAs (miRNAs) will be critical to understanding the role these non-coding RNAs play in regulating gene function and what impact they have in cancer biology. In this study, we show by a combination of microarray, Northern, and PCR-based analyses that the human miR-34a miRNA is significantly under-expressed in basal-like breast cancer cell lines exhibiting triple receptor negative characteristics, as compared to a non-tumorigenic mammary epithelial cell line control. Introduction of a synthetic mimic of hsa-miR-34a into MDA-MB-231 cells resulted in a substantial alteration in both the mRNA and protein levels of the receptor tyrosine kinase AXL, 48 hours post-transfection. Furthermore, the transfection of the hsa-miR-34a mimic decreased the migratory potential of these cells in a manner consistent with the function of AXL as a promoter of cell proliferation, invasion, and metastasis in a number of cancer cell types including breast cancer. This observed phenotypic effect supports the hypothesis that miR-34a functions as a tumor suppressor, with the direct translational repression of AXL representing just one of many possible targets that this miRNA regulates.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2098.
Abstract 210: Glionitrin A causes apoptosis and DNA damage-induced S phase arrest in human prostate cancer cells
Authors: Kim Young-Joo , Kwon Hak Cheol , Yoo Ji-Hye , Yang Hyun Ok 
Published: February 2011

Abstract
In recent study, we have isolated a new diketopiperazine disulfide, Glionitrin A (GN A), from co-culture broth of a Aspergillus fungal strain KMC901 and a Sphingomonas bacterial strain KMK001, and demonstrated anti-microbial and anti-tumor effects; however, its mechanisms of action are not understood yet. Here, we investigated anti-tumor activity of GN A and its underlying molecular mechanism in human prostate cancer DU145 cells in vitro and in vivo. GN A showed significant cytotoxicity (IC50, 1.5 μM) against DU145 cells through a caspase-8-dependent apoptotic pathway. GN A treatment (0.75 ∼ 3 μM) activated caspase-8, 9 and 3, and also increased formation of truncated BID and expression of BAX protein in a dose-dependent manner. Furthermore, GN A elevated phospho-H2A.X (Ser139), which is known to be phosphorylated by ATM/ATR in response to DNA damage, and activated S phase checkpoint associated with ATM-Chk1/2-Cdc25A pathway. In vivo study performed in nude mice bearing xenografts of DU145 (1×107 cells/mouse, s.c.), GN A dramatically reduced the tumor volume by an average of 38.2% (5 mg/kg, p.o.) and 71.3% (10 mg/kg, p.o.) at the 27-day time point, respectively. These observations may provide aid in better understanding of the mechanism underlying the anti-tumor activity of GN A, which has the potential to be developed into an anti-prostate cancer agent.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 210.
Abstract 2106: Regulation of Rb1 and caspase 3/7 by miR-17-5p, miR-132/-212, and miR-337-3p: Oncogenic role of miRNAs in pancreatic cancer
Authors: Park Jong Kook , Jiang Jinmai , Gusev Yuriy , Lerner Megan R. , Brackett Daniel J. , Schmittgen Thomas D. 
Published: February 2011

Abstract
Pancreatic ductal adenocarcinoma (PDAC) continues to be one of the most lethal forms of cancer. Reasons for the high mortality associated with this disease include a lack of effective treatments and poor early detection methods. Enhanced understanding of the pathways that are deregulated in PDAC could lead to improved therapies and diagnostics. We and others have reported a microRNA expression signature that is associated with PDAC. The purpose of this study was to identify target genes and pathways that are regulated by miRNAs with altered expression in PDAC. We found several miRNAs including miR-17-5p, −132/-212, and −337-3p that have increased expression in clinical specimens of PDAC compared to normal and adjacent benign pancreas. miR-132 and −212 are located within 472 bp of each other on chromosome 17, have the identical seed sequence and should therefore regulate the identical targets. Through a luciferase reporter assay and western blotting, we found that miR-132/-212 directly regulate the expression of the tumor suppressor Rb1 in the pancreatic tumor cell lines HS766T and PANC-1. Over expression of these miRNAs (miR-132/-212) either through stable expression or pre-miR oligos increased the proliferation rate of HS766T and PANC-1 cells. Expression of 18 genes that are transcriptionally regulated by the E2F including Cyclin A, Cyclin B, PCNA, MCM2, MKI67 and others was increased following miR-132/-212 over expression. Additional genes that are predicted to be targeted by these miRNAs include procaspase 3 (miR-337-3P) and procaspase 7 (miR-132/-212 and miR-17-5p). These caspases are essential in executing programmed cell death in tumors. Western blotting and luciferase reporter assays determined that procaspase 3 and procaspase 7 are regulated by miR-337-3P and miR-132/-212 & miR-17-5p, respectively. Our data strongly suggest that over expression of miR-132/-212 contributes to the enhanced proliferation that is observed in pancreatic cancer. Furthermore, this study is the first to discover that main effector caspases, caspase 3 and 7, are post-transcriptionally regulated by miRNAs. Our findings suggest that resistance to chemotherapeutics in pancreatic tumor tissues may be linked to reduced expression of these procaspases.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2106.
Abstract 2110: Comparing detection of EGFR mutations in lung cancer patients
Authors: Ikeda Takaya , Nakatomi Katsumi , Yamaguchi Hiroyuki , Nakamura Yoichi , Ogawara Taiki , Senju Hiroaki , Tomonaga Nanae , Doi Seiji , Iida Tetsuya , Igawa Tsukasa , Nagayasu Takeshi , Tsukamoto Kazuhiro , Kohno Shigeru 
Published: February 2011

Abstract
INTRODUCTION:
Epidermal growth factor receptor (EGFR) mutations are a strong determinant factor of response to EGFR tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC). Now some methods have been used to detect EGFR mutations and they have approximately the same sensitivity and specificity. In a case, we experienced unmatched results from two methods of detecting EGFR mutations. In this study, we compared data from three methods of detecting mutations.
METHODS:
We obtained 30 samples of operation, transbronchial lung biopsy, bronchoscopic brushing, pleural effusion and lymph nodes from NSCLC patients. EGFR mutation status was determined by two or three methods (mutant-enriched PCR method, PNA-LNA PCR Clamp method and PCR invader method).
RESULTS:method (n=31)mutation positive2 method (n=20)m.e./Clamp (n=10)4/4 m.e./invader (n=2)1/0 Clamp/invader (n=8)4/33 method (n=10)m.e./Clamp/invader (n=10)3/2/3m.e.: mutant-enriched PCR, Clamp: PNA-LNA PCR Clamp, invader: PCR invader
EGFR mutations were detected in 12 samples. Mutation statuses of two or three methods were matched in 9 samples, but 3 samples were unmatched.
CONCLUSIONS:
EGFR mutation statuses were unmatched in same samples. EGFR gene status is essential information for NSCLC patients. We need to take false negative into consideration in the clinical setting and need father deliberation about detecting methods of EGFR mutations.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2110.
Abstract 2113: A wild-type-blocking reference sequence enhances COLD-PCR and enables fast amplification and high enrichment of all types of low-prevalence unknown mutations
Authors: Milbury Coren A. , Li Jin , Makrigiorgos G. Mike 
Published: February 2011

Abstract
Background: Molecular profiling of somatic mutations in cancer often requires the identification of low-prevalence DNA mutations in an excess of wild-type (WT) DNA; however, the method selectivity and sensitivity are often limiting factors. COLD-PCR (CO-amplification at Lower Denaturation Temperature) resolves several limitations in low-level mutation detection by using critical denaturation temperatures to enrich mutant-containing amplicons during PCR. However, certain mutation types are not enriched by some COLD-PCR formats. We report a novel enhancement in COLD-PCR that enables an Improved and Complete Enrichment (ice-COLD-PCR) for all mutation types.
Methods: A Reference Sequence (RS) that preferably forms double-stranded structures with WT sequences, but not with mutant sequences, was added to COLD-PCR. A RS was designed to bind to one WT-sequence strand, avoid primer binding and prevent polymerase extension. To validate the use of a RS in COLD-PCR, we evaluated segments of TP53 exon 8. Serial dilutions of mutant cell-line DNA, or human lung tumor DNA, in WT DNA were analyzed. Mutations that increase, decrease, or retain the amplicon melting temperature were tested. Following conventional-PCR, COLD-PCR (full or fast COLD-PCR formats), and ice-COLD-PCR methods amplicons were sequenced and the degree of mutation enrichment was compared. Several clinical lung adenocarcinoma samples with known low-level mutations were also tested with ice-COLD-PCR.
Results: ice-COLD-PCR yielded ∼13-fold enrichment for Tm-increasing and Tm-equivalent mutations, and ∼15-fold enrichment for Tm-reducing mutations. In contrast, Full-COLD-PCR demonstrated ∼5-fold enrichment for all mutations. Further, fast-COLD-PCR, which can only enrich Tm-reducing mutations, exhibited ∼17-fold enrichment for these types of mutations, while the Tm-increasing and Tm-equivalent mutations remained undetectable. Regardless of mutant type and position, after ice-COLD-PCR amplification, all mutation types are strongly enriched and can be reliably sequenced down to a level of 1-3%. Ice-COLD-PCR duration is ∼1hour, compared to several hours for full-COLD-PCR.
Conclusions: The inclusion of appropriately designed RS within COLD-PCR selectively inhibits WT amplification throughout PCR, while preferentially enriching mutants and reducing time-intensive hybridization times. Ice-COLD-PCR combines high sensitivity, speed, and low-cost, and facilitates direct sequencing for all types of unknown low-prevalence mutations in clinical cancer samples.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2113.
Abstract 2118: Rare allele enrichment by ARMS combined with probe base genotyping techniques
Authors: Zhou Luming , Smith Denice , Wittwer Carl 
Published: February 2011

Abstract
Mutation detection plays a key role in several medical areas for diagnosis, treatment and prognosis. Additionally, the ability to detect low level mutations among mostly wild-type DNA is critical to areas such as cancer detection, prenatal testing and infectious diseases. For example, low-level (&lt;10%) cancer mutations can not be sequenced by standard PCR or genotyping techniques. Many moderate- to high-selectivity PCR methods have been developed over the past two decades to enrich minority alleles for known mutations. One of the most widely used approaches is the amplification refractory mutation system (ARMS) which relies on the use of 3’ end terminal nucleotide mismatch to enhance the minor allele. The mutation detection sensitivity ranges from 1:10 to 1:1000. Probe based genotyping techniques such as fluorescent labeled probes, unlabeled probes and Snapback primers depend on short oligonucleotides that hybridize to the template for genotyping. We have combine allele enrichment by ARMS and probe based genotyping techniques to enhance allele discrimination. We designed a 3’ end blocked oligonucleotides to form a perfect match with the wild- type allele (mismatched to other alleles). Therefore, the probe blocks the primer from annealing to the wild-type template, while simultaneously permitting the primer to anneal to, and amplify, the mutant template. Wild-type amplification is completely blocked, and the mutation is enriched. The sensitivity of unlabeled probe-ARMS mutation enrichment detection is 1:30,000. Combining ARMS enrichment of minority alleles and snapback primers has the additional benefit that only two PCR primers are required, with the addition of a short tail of nucleotides on one primer that results in an intra-molecular hybridization probe. The sensitivity of snapback-ARMS mutation enrichment detection is 1:60,000. Fourty-four patients with paired thyroid biopsies and needle samples and 3 additional patients with only needle samples were tested for the BRAF mutation p.V600E (c.1799 T&gt;A) by dual hybridization probes. A blinded test using unlabeled probe-ARMS and Snapback-ARMS enrichment techniques were compared to the dual hybridization probe method. All samples were concordant (47 positive and 42 negative) except 4 samples were positive using unlabeled probe-ARMS and Snapback-ARMS but were negative using hybridization probes. Minority allele enrichment using ARMS combined with probe base genotyping methods increases the mutation detection sensitivity 100 times or more. ARMS combined with an unlabeled probe or a snapback primer is an inexpensive and rapid enrichment technique. No separation, purification or addition steps that increase risk of contamination are necessary.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2118.