|ognizant Communication Corporation|
Featuring PRECLINICAL AND CLINICAL CANCER THERAPEUTICS
VOLUME 18, NUMBER 9
Oncology Research, Vol. 18, pp. 401-408
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Distinct DNA Methylation Profiles Between Adenocarcinoma and Squamous Cell Carcinoma of Human Uterine Cervix
Eun-Ju Lee,1,2 Michael McClelland,1 Yipeng Wang,1,3 Fred Long,1 Sang-Ho Choi,1,4 and Je-Ho Lee5,6
1Vaccine Research Institute of San
Diego, San Diego, CA, USA
2Department of Obstetrics and Gynecology, Chung-Ang University School of Medicine, Seoul, Korea
3Pathology and Laboratory Medicine, University of California at Irvine, Irvine, CA, USA
4Department of Infectious Diseases, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
5Molecular Therapy Research Center, Sungkyunkwan University, Seoul, Korea
6Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
Alterations in DNA methylation offer unique prospects as tumor markers. The big limitation in cervical cancer research is that it is too hard to obtain the pure normal tissue from a cervical cancer mass. So, we first profile type-specific DNA methylation of major two types of human uterine cervical cancer, adenocarcinoma (ACA) and squamous cell carcinoma (SCC), to establish a precise source of marker research. To assess the DNA methylation status of promoter regions in human uterine cervical ACAs and SCCs, fresh frozen tissues were obtained from bulky tumor masses to minimize the contamination from normal tissues and two array platforms using digestion with methylation-sensitive restriction-enzyme HpaII, ligation, and PCR were performed: an array of 11,994 (~1.5 kb) PCR products from 10,445 promoter regions, and an array of 355,264 oligonucleotides for 18,212 HpaII fragments in 12,617 promoter regions. Loci near 21 genes showed significant differences between six ACA and four SCC from the analysis of two array data. Real-time PCR-based validation was performed on 13 loci using other nearby candidate methylation targets in the same promoter. Methylation patterns of 11 of 13 linked loci concurred with the microarray results. Four loci were further studied using tissues from additional patients (23 ACA and 24 SCC). Hypermethylation of loci in PAK6 and NOGOR most strongly correlated with ACA. Therefore, we have identified the 21 genes with differential methylation pattern between ACA and SCC and, furthermore, we found that PAK6 and NOGOR could be useful markers of ACA to be distinct from SCC.
Key words: DNA methylation; Cervical cancer; PAK6; NOGOR
Address correspondence to Je-Ho Lee, M.D., Ph.D., Molecular Therapy Research Center, Sungkyunkwan University, Samsung Cancer Center, Ilwondong, Kangnamgu, Seoul, 135-710, Korea. Tel: 82-2-3410-3510; Fax: 82-2-2148-7329; E-mail: email@example.com
In Vivo Effect of a-Bisabolol, a Nontoxic Sesquiterpene Alcohol, on the Induction of Spontaneous Mammary Tumors in HER-2/neu Transgenic Mice
Laura Costarelli,1 Marco Malavolta,1 Robertina Giacconi,1 Catia Cipriano,1 Nazzarena Gasparini,1 Silvia Tesei,1 Sara Pierpaoli,1 Fiorenza Orlando,2 Hisanori Suzuki,3 Luigi Perbellini,4 Francesco Piacenza,5 Monica Emanuelli,6 and Eugenio Mocchegiani1
1Center on Nutrition and Ageing, INRCA,
2Experimental Animal Models for Ageing Unit, INRCA, Ancona, Italy
3Department of Neuroscience and Vision, Section of Biochemistry, University of Verona, Verona, Italy
4Department of Medicine and Public Health, University of Verona, Verona, Italy
5Department of Molecular Pathology and Innovative Therapies, Occupational Medicine, Polytechnic University of Marche, Torrette, Ancona, Italy
6Institute for Biochemical Biotechnologies, Polytechnic University of Marche, Ancona, Italy
Breast cancer represents the most commonly diagnosed invasive malignancy in pre- and postmenopausal women in both developed and underdeveloped countries. Taking into account that treatment options, including surgery, have not been able to deal with the growing incidence of breast malignancy, it is required to develop mechanism-based novel agents for its prevention. Wide interest in some natural compounds as anti-inflammatory agents and as alternative to the traditional medicines is increasing because they do not provoke any adverse effects and are effective in multiple organs. a-Bisabolol (BISA), a small oily sesquiterpene alcohol, was reported as chemopreventive agent in induced rat mammary carcinogenesis. The aim of the present study is to investigate the role played by two doses of BISA (via intramammary infusion) on the induction and development of mammary tumor in HER-2/neu transgenic mice as well as the BISA effect after tumor surgical resection. The main data show that (a) optimal dosage of BISA is 10 mg/mouse rather than 3.6 mg/mouse with no adverse effects (e.g., alopecia); (b) the number of the palpable tumor masses decreases in mice treated with 10 mg/mouse of BISA; (c) mice after surgical resection of the primary tumor and treatment with BISA (10 mg) are free from tumor for more weeks, after the surgical treatment; (d) using array analysis, some genes implicated in carcinogenesis mechanisms (NF-kBia, Map2k, Mapk14, and HER2/ neu), angiogenesis process (Fgf), and inhibition of apoptosis (Birc5) are differently regulated after BISA treatment, with a downregulation of the HER2/neu as well as of Fgf and Birc5 genes; (e) the NK cell cytotoxicity increases in tumor-treated mice, especially after the removal of the first tumor mass. Such effectiveness could be important to achieve goals for a possible combintion of BISA to conventional therapies in breast cancer and to tumor surgical removal (adjuvant therapy), as suggested for other sesquiterpene analogs.
Key words: Natural substances; Breast cancer; HER-2/neu; Array analysis; Adjuvant therapy
Address correspondence to Dr. Eugenio Mocchegiani, Center on Nutrition and Ageing, PST-INRCA, Via Birarelli 8, 60121, Ancona, Italy. Tel: +39-071-8004216; Fax: +39-071-206791; E-mail firstname.lastname@example.org
The Importance of Release of Proinflammatory Cytokines, ROS, and NO in Different Stages of Colon Carcinoma Growth and Metastasis After Treatment With Cytotoxic Drugs
Roman Paduch,1 Martyna Kandefer-Szerszen,1 and Tomasz Piersiak2
1Department of Virology and Immunology,
Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University,
2Department of Comparative Anatomy and Anthropology, Institute of Biology, Maria Curie-Sklodowska University, Lublin, Poland
In colorectal cancers, the local cytokine network and the levels of nitric oxide (NO) and reactive oxygen species (ROS) are known to be closely related to cancer progression and metastasis, but the influence of the currently administered therapies on the cancer microenvironment is not completely understood. We analyzed the levels of reactive oxygen species (ROS), nitric oxide (NO), and cachexia-mediated cytokines (IL-1b, IL-6, TNF-a) in cocultures of human colon carcinoma spheroids prepared with cells derived from tumors of different grades with human normal colon epithelial and myofibroblast cells and normal endothelial cells. We also analyzed the influence of standard chemotherapy with 5-fluorouracil (5-FU) and leucovorin (LV) combined with camptothecin (CPT-11) (IFL regimen with drug concentrations adjusted to in vitro conditions) on these parameters. The results indicated that adhesion of colon carcinoma spheroids to colon epithelium and myofibroblast monolayers induced O2- anion production but decreased NO levels compared to the sum of the radicals released by monocultures of the two types of cells. Coculture of colon carcinoma spheroids with endothelium was an exception to this rule, as only HT29 cells decreased NO production. In cocultures, anticancer drugs additionally, though only slightly and insignificantly, increased the production of the radicals compared to a nontreated coculture, but in monocultures, the drugs, and especially CPT-11, were ROS inducers and simultaneously NO production inhibitors. However, the levels of released ROS and NO were dependent on the stage of colon carcinoma that the cells were derived from. LS180 cells (grade B) grown in monocultures produced the lowest ROS levels but were the best producers of NO. Adhesion of tumor spheroids to normal cells influenced the microenvironmental cytokine network compared to monocultures, decreasing IL-1b and TNF-a secretion but significantly enhancing IL-6 levels. The addition of the drugs had no effect on IL-1b levels but increased TNF-a production and lowered the amounts of IL-6. In conclusion, cytotoxic drugs may, dependent on the stage of tumor growth or the type of chemotherapy regimen administered, significantly influence the proinflammatory cytokine network and local ROS and NO levels. Moreover, in cocultures of tumor cells with normal epithelial, myofibroblast, and endothelial cells, ROS production seems to be involved in local cell injury, which was detected by confocal microscopy. On the other hand, high level of NO seems to facilitate tumor cell interactions with the endothelium and metastasis as NO production was the highest in a monoculture of HUVEC and remained at high levels in cocultures of colon cancer cells with HUVEC. Among the proinflammatory cytokines, only IL-6 seems to significantly influence colon carcinoma development and metastasis. Attenuation of IL-6 production after chemotherapy can be a useful prognostic factor of its effectiveness.
Key words: Colon carcinoma; Proinflammatory cytokines; Cytotoxic drugs; Coculture; Reactive oxygen species; Nitric oxide
Address correspondence to Dr. Roman Paduch, Department of Virology and Immunology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin, Poland. Tel/Fax: +48 81 537 59 40; E-mail: email@example.com
Meta-Analysis of Chemotherapy With Irinotecan or Oxaliplatin-Involved Regimen for Untreated Metastatic Advanced Colorectal Cancer
Luhong Zhuang,1* Jianling Bai,2* Huaying Huang,1 Cuiju Tang,1 Jinsong Yang,1 Baoning Zhou,1 Yongling Gong,1 Zhong Duanmu,1 and Jinfei Chen1
1Department of Oncology, The Affiliated
Nanjing First Hospital, Nanjing Medical University, Nanjing, P.R. China
2Department of Epidemiology and Biostatistics, Nanjing Medical University, Nanjing, P.R. China
A large number of randomized controlled trials involving chemotherapy in the management of advanced colorectal cancer were conducted. 5-FU/LV in combination with irinotecan (IRI) or oxaliplatin (OXA) was used. The aim of the meta-analysis was to compare and evaluate the effectiveness and safety of the two therapeutic approaches for patients with advanced colorectal cancer. A literature search, study selection and assessment, data collection, and analysis were undertaken by two reviewers according to the Cochrane Handbook for Systematic Reviews of Interventions. Randomized controlled trials (RCTs) or quasi-RCTs comparing IRI versus OXA, in combination with 5-FU/LV in the treatment of advanced colorectal cancer were performed. Seven studies involving 2,107 patients met the inclusion criteria. The OXA + 5-FU/LV regimen showed a significant increase in survival by lower hazard ratios (HR) [HR 1.28; 95% CI (1.13-1.45)] and was associated with lower toxicities. The OXA + 5-FU/LV regimen was superior or equal to the IRI + 5-FU/LV regimen in prolonging time to progression and median survival. The IRI + 5-FU/LV regimen resulted in higher hazard ratios in nausea vomiting/emesis and diarrhea [HR 1.99, 95% CI (1.19-3.31); HR 1.83, 95% CI (1.38-2.44)] and lower hazard ratios in paresthesia, sensory neuropathy, and thrombocytopenia [HR 0.09, 95% CI (0.03-0.23); HR 0.04 95% CI (0.01-0.13); HR 0.19 95% CI (0.05-0.64)] than the OXA + 5-FU/LV regimen. Compared with IRI, OXA is more appropriate for the treatment of advanced colorectal cancer when combined with 5-FU/LV. OXA + 5-FU/LV should be considered as the first-line standard of care for advanced CRC patients.
Key words: Colorectal cancer; Chemotherapy; Irinotecan; Oxaliplatin; Systematic review; Meta-analysis
Address correspondence to Jinfei Chen, M.D., Ph.D., The Affiliated Nanjing First Hospital, Nanjing Medical University, Nanjing, P. R. China, 210006. Tel: 0086-025-52271251; Fax: 0086-025-52271251; E-mail: firstname.lastname@example.org
*These authors contributed equally.
Prognostic Value of E-Cadherin Expression in Non-small Cell Lung Cancer Treated With Gefitinib
Eun Hee Kook,1 Yeo Myeong Kim,1 Hyeon Tae Kim,1 Jae Soo Koh,2 Yun Jung Choi,1 Jin Kyung Rho,1 Hye-Ryoun Kim,1 Cheol Hyeon Kim,1 and Jae Cheol Lee1
1Department of Internal Medicine, Korea
Cancer Center Hospital, Seoul, Korea
2Department of Pathology, Korea Cancer Center Hospital, Seoul, Korea
E-cadherin expression has been suggested to be related with response to EGFR-TK inhibitors. To evaluate the prognostic value of E-cadherin expression in lung cancer treated with gefitinib, we retrospectively reviewed patients with inoperable stage IIIb or IV non-small cell lung cancer treated with gefitinib and compared immunohistochemical staining for E-cadherin on biopsied specimens. The association between gefitinib sensitivity and E-cadherin expression by Western blot and immunocytochemistry was also examined in 10 lung cancer cell lines. Of 52 eligible patients, 15 (28.8%) showed a partial response, and the disease control rate was 42.3%. Median progression-free survival (PFS) and overall survival (OS) in responders were 8 and 20 months, respectively. Response rates (RR), PFS, and disease control rates were higher in never-smokers and patients showing adenocarcinoma histology, and improved OS was observed in patients with adenocarcinoma. E-cadherin expression did not impact any parameters (RR, PFS, or OS). Although there was a tendency for cell lines with lower IC50 to have E-cadherin expression, H2009 cells were the least sensitive to gefitinib, with an IC50 of 30 mM, and H1650 cells had an intermediate sensitivity despite high E-cadherin expression. E-cadherin expression was not a useful indicator of response and survival after treatment with gefitinib.
Key words: E-cadherin; Epidermal growth factor receptor; Gefitinib; Lung cancer
Address correspondence to Jae Cheol Lee, Department of Internal Medicine, Korea Cancer Center Hospital, 215-4, Gongneung-dong, Nowon-gu, Seoul 139-706, Korea. Tel: +82 2 9701206; Fax: +82 2 9702410; E-mail: email@example.com
p53 Codon 72 and p21 Codon 31 Polymorphisms and Susceptibility to Cervical Adenocarcinoma in Korean Women
Ju-Won Roh,1 Bu Kyung Kim,2 Chae Hyeong Lee,1 Jongseung Kim,3 Hyun Hoon Chung,4 Jae Weon Kim,4 Noh-Hyun Park,4 Yong-Sang Song,4 Sang-Yoon Park,2 and Soon-Beom Kang4
1Department of Obstetrics and Gynecology,
Dongguk University, Goyang, Korea
2Center for Uterine Cancer, Research Institute and Hospital, National Cancer Center, Goyang, Korea
3Department of Family Medicine, Boramae Medical Center, Seoul Metropolitan Government Seoul National University, Seoul, Korea
4Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea
The aims of this study were to evaluate the genotype frequencies of p53 codon 72 and p21 codon 31 in cervical adenocarcinoma patients and controls, and the association between the specific genotype or genotype combination of these polymorphisms and the risk of cervical adenocarcinoma in Korean women. Genotyping was performed using DNA from cervical biopsy specimens collected from 53 patients with cervical adenocarcinoma, of whom 34 were HPV 16 or 18 positive, and from the cervical exfoliated cells from 286 control women, of whom 48 were positive for HPV 16 or 18. For the determination of p53 polymorphisms genomic DNA was examined by PCR amplification of the specific allele assay, and for the determination of p21 polymorphisms DNA was examined by the PCR-RFLP assay using BsmAI. We found significant differences in genotype frequencies of both genes between the two groups (p < 0.001). The p53 genotypes containing the Pro allele were significantly associated with cervical adenocarcinoma with an OR of 2.89 (95% CI 1.54-5.42). Also, homozygous carriers of the p21 Ser allele showed a substantially increased risk of developing cervical adenocarcinoma (OR 2.07; 95% CI 1.13-3.79) compared to genotypes containing the Arg allele. In addition, the combination of the Pro allele containing genotypes of p53 and the Ser homozygous genotype of p21 posed a remarkably increased risk (OR 5.22; 95% CI 2.24-12.16), although the interaction of the two genes could not be found. These significant differences were intensified in groups with high-risk HPV infection (types 16 or 18).
Key words: p21 codon 31; p53 codon 72; Polymorphism; Adenocarcinoma; Cervical cancer
Address correspondence to Ju-Won Roh, Department
of Obstetrics and Gynecology, Dongguk University Ilsan Hospital, 410-773
Goyang, Korea. Tel: +82 31 961 7363; Fax: +82 31 961 7977; E-mail: firstname.lastname@example.org
or Jae Weon Kim, Department of Obstetrics and Gynecology, Seoul National
University College of Medicine, Seoul 110-744, Korea. Tel: +82 2 2072 3511;
Fax: +82 2 762 3599; E-mail: email@example.com