|ognizant Communication Corporation|
Featuring PRECLINICAL AND CLINICAL CANCER THERAPEUTICS
VOLUME 17, NUMBER 10
Oncology Research, Vol. 17, pp. 437-445
0965-0407/09 $90.00 + .00
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Potentiation of Proliferation of Some But Not All Human Colon Carcinoma Cell Lines by Immobilized Hepatic Asialoglycoprotein Receptor 1
Jinbo Fang,* Ryota Izawa, Laura Gomez-Santos, Suguru Ueno, Tomoya Sawaguchi, Katsuaki Usami, Yoshiaki Nodera, Hideyuki Takeuchi, Yoshimi Ohashi, Nobuaki Higashi, and Tatsuro Irimura
Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan
Twenty-three human colorectal carcinoma cell lines were examined for the binding of recombinant hepatic asialoglycoprotein receptor 1 (ASGR1), which is known to be exclusively expressed on hepatic parenchymal cells. The effects of the binding were assessed by adhesion to and proliferation on immobilized recombinant ASGR1. Recombinant ASGR1 bound strongly to six cell lines and moderately to 15 cell lines out of 23 lines tested, as shown by flow cytometric analysis. The first six cell lines (group A) also exhibited strong adherence to immobilized ASGR1, whereas 11 of the 15 cell lines of the second group (group B) showed significant adhesion with smaller enhancement by ASGR1 than the cell lines in group A. With a representative cell line (DLD-1 cells categorized in group B), a significant portion of the adhesion was inhibited by preincubation of ASGR1 with asialofetuin, a competitive inhibitor of the carbohydrate recognition by ASGR1. The growth rates of 13 cell lines (two of group A and 11 of group B) were significantly accelerated when they were cultured on immobilized recombinant ASGR1. The results indicate that ASGR is a potential organ-specific microenvironmental factor for colorectal carcinoma growth and metastasis formation in livers.
Key words: Hepatic asialoglycoprotein receptor; Colorectal carcinoma cells; Liver metastasis; Organ microenvironment
Address correspondence to Dr. Tatsuro Irimura, Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-0033, Japan. Tel: +81-3-5841-4870; Fax: +81-3-5841-4879; E-mail: email@example.com
*Present address: School of Life Sciences, Northeast Normal University, Changchun 130024, Jillin Province, P.R. China.
Bcl-2 in Combination to Myeloid Antigen Expression in Adult Acute Lymphoblastic Leukemia and Prognostic Outcome
Zahra Amirghofran,1,2 Yahya Daneshbod,3 and Naser Gholijani1
1Department of Immunology, Shiraz University
of Medical Sciences, Shiraz, Iran
2Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
3Department of Pathology, Shiraz University of Medical Sciences, Shiraz, Iran
The present study was performed to find the importance of two myeloid (CD13 and CD33) antigens aberrantly expressed on the blasts of acute lymphoblastic leukemia (ALL) patients and Bcl-2 expression in relation to clinical and biological features and treatment outcome. Bone marrow or peripheral blood samples of 50 patients were assessed for the expression of markers by immunostaining methods. Twenty-one patients (42%) showed more than 20% positivity for Bcl-2. Aberrant expression of myeloid antigens was found in 14% of cases. The expression of Bcl-2 was associated with shorter survival (p = 0.009). A significant correlation between expression of myeloid antigens (MY) and survival and complete remission duration was found. The mean survival was 656 ± 301 days for MY+ cases and 1,009 ± 230 days for MY- patients (p < 0.0001). Expression of Bcl-2 in combination to myeloid antigens was associated with a poorer outcome. Survival of MY+ patients expressing Bcl-2 was shorter than MY- Bcl-2+ and MY+ Bcl-2- ALL cases (p = 0.038). In conclusion, results of this study indicated the prognostic value of Bcl-2 and myeloid antigen expression in ALL patients. Presence of these markers together on the leukemic cells was associated with a poorer response to therapy and may implicate modified therapeutic strategies in the patients.
Key words: Acute lymphoblastic leukemia; Bcl-2; Myeloid antigens
Address correspondence to Zahra Amirghofran, Ph.D., Department of Immunology, Medical School, Shiraz University of Medical Science, 71345-1798, Shiraz, Iran. E-mail: firstname.lastname@example.org
Effects of Genomic Imbalances on Telomerase Activity in Gastric Cancer: Clues to Telomerase Regulation
Güvem Gümüs-Akay,1 Atilla Halil Elhan,2 Ali Ekrem Ünal,3 Ahmet Demirkazik,4 Asuman Sunguroglu,1 and Ajlan Tükün5
1Department of Medical Biology, Ankara
University Faculty of Medicine, Ankara, 06100, Turkey
2Department of Biostatistics, Ankara University Faculty of Medicine, Ankara, 06100, Turkey
3Department of General Surgery, Division of Surgical Oncology, Ankara University Faculty of Medicine, Ankara, 06100, Turkey
4Department of Medical Oncology, Ankara University Faculty of Medicine, Ankara, 06100, Turkey
5Department of Medical Genetics, Ankara University Faculty of Medicine, Ankara, 06100, Turkey
Telomerase is a specialized cellular reverse transcriptase that adds telomeric repeats (TTAGGG) at the ends of each chromosome. Nearly the complete spectrum of human cancers has been shown to be telomerase positive. The understanding of the telomerase regulation in concert with other genetic alterations in the process of malignant transformation of human cells has important clinical and practical implications. Regulation of telomerase activity (TA) is highly complex, and both putative positive and negative regulators have been reported. However, the mechanisms involved in telomerase regulation are not fully established. Identification of additional telomerase components and associated proteins will certainly contribute to further investigations of the effect of telomerase in telomere elongation, telomere length maintenance, oncogenesis, and functionally new, unidentified cellular functions. In this study our aim was to determine the chromosomal localizations of putative unidentified telomerase activator(s) and/or repressor(s) by high resolution-comparative genomic hybridization (HR-CGH) in highly telomerase expressing gastric tumor samples. For this purpose TAs and genomic imbalances were identified in the same tumor samples and relation between these was evaluated. Genomic changes affecting telomerase activity in 50 gastric tumor samples were investigated by HR-CGH. We have found that genomic imbalances including 1q+, 8p+, 8q+, 10q+, 17p-, and 20p+ are associated with the higher telomerase activity. Our results suggest that 1q24, 8p21-p11.2, 8q21.1-q23, 10q21-qter and 20pter-p11.2 may contain putative telomerase activator(s), whereas the 17p12 region may harbor candidate telomerase suppressor(s).
Key words: Telomerase activity; Telomerase regulation; HR-CGH; Genomic imbalances; Gastric cancer
Address correspondence to Prof. Dr. Ajlan Tu¨ku¨n, M.D., Ph.D., Department of Medical Genetics, Ankara University Faculty of Medicine, 06100 Sihhiye/Ankara, Turkey. Tel: +90 312 310 30 10 /351; Fax: +90 312 310 63 70; E-mail: Ajlan.Tukun@medicine.ankara.edu.tr
Hepal-6 Derived RNA Electroporated Murine Spleen B Cells Induced Antitumor Effects In Vivo
SuNan Shen, YanFei Xu, and Feng Yu
Jiangsu Key Laboratory for Molecular Medicine, Medical School of Nanjing University, Nanjing, P.R. China
RNA electroporated CD40 ligand-activated B cells can induce cytotoxic T-lymphocyte response in vitro. In order to evaluate the effects in vivo, we applied murine spleen B cell vaccine in a mouse model of liver cancer. C57BL/6 mouse spleen B cells were activated by anti-mouse CD40 antibody, recombinant interleukin-4, and cyclosporin A, and then electroporated with total RNA from Hepal-6 cells (Hepal-6 RNA-CD40-mAb-B cells). This vaccine was injected into C57BL/6 mice that were subcutaneously inoculated with Hepal-6 cells. Hepal-6 RNA-CD40mAb-B cells could induce tumor-specific cytotoxic T cells and IFN-g secretion in the immunized mice. In vivo study showed this vaccine could well inhibit tumor progression, improve overall survival, and provide continuous immunoprotection against Hepal-6 cell-induced tumor. Tumor cell-derived total RNA electroporated B cells vaccine is a type of effective immunotherapy and provides potential implication for clinical treatment.
Key words: CD40; B cell; Immunotherapy; Hepatocellular carcinoma; C57BL/6 mouse
Address correspondence to SuNan Shen, M.D., Ph.D., Jiangsu Key Laboratory for Molecular Medicine, Medical School of Nanjing University, Nanjing, 210093, P.R. China. Tel: +86-25-83686043; Fax: +86-25-83686559; E-mail: email@example.com
Establishment and Characterization of MACL-1 and MGSO-3 Cell Lines Derived From Human Primary Breast Cancer
C. R. Correa, C. M. Bertollo, and A. M. Goes
Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Brazil
Breast cancer is a major health burden worldwide. It is responsible for over 1 million of 10 million cases of cancer in the world. Advances in breast cancer detection and treatment have contributed to improve the rate of survival, although mortality rates remains significantly high. Despite all these advances, more efficient diagnostic methods and effective treatments are necessary. The establishment of breast cancer cell lines is an important tool to understand biological processes involved in this disease, as well as the identification of potential therapeutic targets. In the present work, two cell lines, MACL-1 and MGSO-3, were established from human primary breast cancer based on differential centrifugation, followed by growth in culture for over 70 passages. Characterization of the cell lines included morphology analysis, determination of doubling time, telomerase expression, tumor antigen expression, colony formation in soft agar, and xenograft implantation into nude mice. Morphological examination demonstrated a typical epithelial morphology and PCR analyses showed that both cell lines were telomerase positives. Moreover, MACL-1 and MGSO-3 were capable of growing in soft agar culture, which suggests its metastatic potential, and both demonstrated a positive tumorigenic potential in nude mice. These experimental models open new perspectives on the investigation of breast cancer pathobiology.
Key words: Breast cancer; Establishment; Characterization; Cell line
Address correspondence to Dr. A. M. Goes, Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, UFMG, Caixa Postal 486, CEP 31.270-901, Belo Horizonte, MG, Brazil. Tel: +55-31-3409-2639; Fax: +55-31-3409-2614; E-mail: firstname.lastname@example.org
Evaluation of Molecular Markers in a Rat Model of Mammary Carcinogenesis
G. Vinothini, R. Senthil Murugan, and S. Nagini
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamil Nadu, India
We sought to evaluate the molecular markers involved in breast tumorigenesis in a rat model that mimics many essential elements of human breast cancer. Female Sprague-Dawley rats were divided into two groups. Animals in group 1 were given a single dose of 7,12-dimethylbenz[a]anthracene (DMBA) (20 mg/rat) dissolved in 1 ml of sesame oil by intragastric intubation. Group 2 animals received basal diet and served as control. We analyzed DMBA-induced changes in the expression of CYP isoforms (CYP1A1 and 1B1) involved in DMBA metabolism, markers of oxidative stress (4HNE, HEL, and 8-OHdG), cell survival and proliferation (PCNA, NF-kB-p50, NF-kB-p65, GST-P, and p53), apoptosis (Bcl-2, Bax, caspases, Apaf-1, cytochrome C, and Fas), invasion (uPA, MMP-2, MMP-9, TIMP-2, and RECK), and angiogenesis (VEGF, VEGF-R1, HIF-1a, and PLGF) by immunohistochemical localization, Western blot, and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The present study demonstrates increased carcinogen metabolism, oxidative stress, cell proliferation, together with apoptosis evasion, invasion, metastasis, and neovascularization that may confer a selective growth advantage to DMBA-induced mammary tumors. Aberrant expression of multiple molecules in key signaling pathways in Sprague-Dawley rat mammary tumors renders this model as an important tool for monitoring carcinogenic progression and chemointervention.
Key words: Apoptosis; Breast cancer; Cell proliferation; Oxidative stress
Address correspondence to Dr. S. Nagini, Professor,
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai
University, Annamalainagar-608 002, Tamil Nadu, India. Tel: +91-4144-239842;
Fax: +91-4144-238145/238080; E-mail: email@example.com or snlabau
Clinical and Biological Relationship Between Chronic Lymphocytic Thyroiditis and Papillary Thyroid Carcinoma
Alfredo Antonaci,1 Fabrizio Consorti,1 Stefania Mardente,2 and Gloria Giovannone1
1Department of Surgery "Francesco Durante,"
University "Sapienza" of Rome, Rome, Italy
2Department of Experimental Medicine, University "Sapienza" of Rome, Rome, Italy
The association between chronic lymphocytic thyroiditis and papillary thyroid carcinoma has been investigated for several years from different perspectives but with few attempts to design a common frame of reference to understand the complex mutual interactions between the various pathways of inflammatory response and of thyroid tumor induction and progression. This article reviews the current knowledge and research on this topic according to epidemiologic, immunobiologic, pathologic, and biomolecular points of view, highlighting achievements and lack of knowledge. It draws some conclusions and points at possible future directions for research.
Key words: Chronic lymphocytic thyroiditis; Papillary thyroid carcinoma; Oncogenes; Apoptosis
Address correspondence to Fabrizio Consorti,
Dip. Chirurgia "Francesco Durante," Viale del Policlinico, 00161 Rome,
Italy. Tel: +39-0649970634; Fax: +39-06491695; E-mail: firstname.lastname@example.org