P-113 Lack of correlation between thymidine phosphorylase expression and tumor-response to docetaxel plus capecitabine in advanced non-small cell lung cancer (NSCLC)

Poster Session I /Molecular ceffs. WISP-I LI and Ki-67 LI were 66.0 f 34.2% (mean & SD) and 22.1 f 23.0% (mean f SD), respectively. Fifty-six patients showed high WISP-1 LI; stained in > 30% of tumor cytoplasms and stronger than macrophages, and 32 patients showed low WISP-1 LI. High WISP-I LI was shown in 48 of 59 adenocarcinomas, 8 of 27 squamous cell carcinomas, and 0 of 2 large carcinomas. Spearman’s correlation test showed a significantly inverse correlation between WISP-l LI and the tumor proliferation fraction assessed by Ki-67 immunostaining (p = -0.302, p=O.O07). WISP-1 LI showed significant correlation with gender, histology, or tumor differentiation (p
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P 112

Significant Up-Regulation of Insulin-Like Growth Factor Binding Protein-4 and -2 by K-ras Activation in Pulmonary Airway Epithelial Cells and Cancers

Hanako Sato’, Takuya Yazawa’, Kenji Hamada”, Hisafumi Yamada-Okabe”, Koji Okudela’, Haruhiko Ishii’, Takaaki Ito’, Masayoshi Kanisawas, Takashi Takahashi4, Hitoshi Kitamura5. ’ Dept of Pathology, Yokohama City University School of Medicine, Yokohama, Japan; ’ Pharmaceutical Research Dept. 4, Kamakura Research Laboratories, Chugai Pharmaceutical Co. Ltd., Kamakura, Japan; 3 Emeritus Professor, Dept. of Pathology Yokohama City University School of Medicine, Yokohama, Japan; 4 Dept. of Molecular Oncology Aichi Cancer Centec Nagoya, Japan; 5 Professor, Dept. of Pathology Yokohama City University School of Medicine, Yokohama, Japan K-ras mutations have been reported to be involved in early stage of tumorigenesis of several human cancers including pulmonary adenocarcinoma. To examine the alteration of expression pattern in association with K-ras gene activation in pulmonary airway epithelial cells, we constructed wild type and oncogenie mutated K-ras expression vectors, transfected into a pulmonary peripheral airway epithelial cell line, HPLlA, obtained stable transfectants (HPLlAK (wild type K-ras-transfected), HPLl A-SW (mutated K-ras-transfected)), and compared the mRNA expression patterns between HPLIA-SW and HPLIA-K or empty vector transfectant (HPLIA-C). The oligonucleotide microarray analysis clarified that insulin-like growth factor binding protein (IGFBP)-4 and -2, which bind to IGF and indirectly function as growth inhibitors, were significantly up-regulated in HPLl A-SW, though the K-ras activation accelerated cellular proliferation. To investigate whether the IGFBP-4 and -2 are generally upregulated by K-ras activation in the pulmonary epithelium including carcinoma, we transfected empty, wild type K-ras, or mutated K-ras expression vector into a bronchial epithelial cell line (NHBE) and a large cell lung cancer cell line (TKB5), obtained stable transfectants (NHBE-C, NHBE-K, NHBE-SW; TKB5-C, TKB5-K, TKB5-SW, respectively), and examined the IGFBP-4 and -2 expression. The RT-PCR, western blot, and K-ras inhibition assay analyses revealed that the K-ras activation up-regulated the IGFBP-4 and -2 both in NHBE and TKB5 cells as well as HPLIA. However, the induction of IGFBP-4 and -2 in TKBS-SW was weak in comparison with that in HPLlA-SW and NHBE-SW. These findings suggest that K-ras activation not only accelerates cell growth but also induces growth inhibitory molecules such as IGFBP-4 and -2 especially in non-neoplastic airway epithelial cells. The pulmonary epithelial cells might lose this growth regulation system, as more aggressiveness is gained probably due to accumulation of genetic abnormalities.

P 113 El

Lack of correlation between thymidine phosphorylase expression and tumor-response to docetaxel plus capecitabine in advanced non-small cell lung cancer (NSCLC)

Ji-Youn Han, Eun Kyung Hong, Dae Ho Lee, Sung Min Yoon, Young Suk Park, Jin Soo Lee. Research institute & Hospital, National Cancer Center, Goyang, Korea

Background: Capecitabine (C) was rationally designed to generate 5fluorouracil (5-FU) preferentially in tumor cells. Thymidine phosphorylase (TP) is the key enzyme that converts C to 5-FU and docetaxel (D) is shown to up reg ulate TP in tumor cells, Theoretically, tumors with high levels of TP expression would be more likely to respond to C. We investigated the relationship between TP expression and tumor response to DC chemotherapy. Method: TP expression in tumor cells was determined by immunohistochemistry in 30 tumor samples available from 39 chemo-na’ive NSCLC patients (pts) who were enrolled in a phase II study of DC chemotherapy. DC chemotherapy consisted of D 36 mg/m’ IV on day 1 and 8 plus C 1,000 mglm’ PO BID on days 1-14 of a 21-day cycle, for a maximum of 6 cycles. TP expression levels were classified as follows; negative (< IO%), low (lo-49%), and high (> 50%).

Biology

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Results: TP expression was negative in 11, low in 5, and high in 14 out of 30 cases. The overall positive TP expression rate was 63%. TP expression was positive in 4/4 (100%) squamous cell carcinomas vs. 15/26 (58%) adenocarcinomas (p=O.26) and 18/26 (69%) stage IV vs. l/4 (25%) stage IIIB (p=O.13). Objective tumor responses were observed in 9/19 (47%) cases with positive TP expression as compared with 7/i 1 (64%) cases with negative TP expression (p=O.23). Conclusions: TP expression was observed in 63% of advanced NSCLC. While DC chemotherapy produced objective responses in more than a half of the cases, there was no relationship between TP expression and tumor response to DC chemotherapy.(Supported by NCC Grant N02C120; Capecitabine was provided by Roche Korea).

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114

The correlation of IL-6 and MCL-1 expression in human paprllomavirus 16/18 infected-lung cancer

Yih-Hsin Chang’, Hong-De Huangs, Ming-Yuh Shiau3, Ya-Wen Cheng4, Huei Lee’. ’ School of Medical Technology, Cbung Shan Medical University, Taichung, Taiwan, Republic of China; 2 institute of Toxicology: Chung Shan Medical Universifl: Taichung, Taiwan, Republic of China; 3 institute of Medicine, Chung Shan Medical University; Hung Kuang University, Taichung, Taiwan, Republic of China; 4 Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan, Republic of China Lung cancer is the leading cause of cancer death in Taiwanese women nonsmokers since 1982. The aetiology of lung cancer for nonsmokers remains unknown. Our previous report demonstrated that human papillomavirus (HPV) 16/18 infection was associated with nonsmoking Taiwanese female lung cancer. Besides, HPV18 E6 and E7 mRNA and proteins can be detected in a half of HPV DNA positive-lung tumors, The above observation suggested that HPV 16/16 infection may play a role in the lung tumorigenesis. Using human cervical cancer cell lines as study models, a previous report indicated that the anti-apoptotic effect of interleukin 6 (IL-6) was mediated by MCI-1 expression and such relationship between IL-6 and MCI-1 was closely associated with HPV infection. In this study, we attempted to reveal whether HPV infection is associated with the induction of MCI-1 and IL-6 expression in lung tumor tissue in vivo. lmmunohistochemistry data showed that 68% and 56% of IL-6 and Mcl1 in 88 lung tumors had positively immunostainings, respectively. A significant association between IL-6 expression and tumor stage was observed (p=O.O43). The expression of IL-6 and MCI-I was significantly associated with HPV 16/18 infection (p=O.O14 and p=O.O04 respectively). We also observed that the expression of IL-6 was significantly correlated with MCI-I expression (p
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Telomerase activation and p16 methylation status in short-term cultures of human bronchial epithelial(HBE) cells

Hveon Woo Yim’, Robbert J.C. Slebos’ , Scott H. Randells, Alden M. Parsons’, M Patricia Rivera*, Frank C. Detterbecks, Jack A. Taylor3. ’ National institute of Environmental Health Sciences, N/H, Research Triangle Park, USA; ’ University of North Carolina at Chapel Hi//, Chapel Hi//, USA; 3 National institute of Environmental Health Sciences, N/H, Research Triangle Park, USA Most lung cancers can be attributed to cigarette smoking where continued exposure of carcinogens results in the accumulation of genetic and epigenetic alterations. Recent studies show that expression of the human telomerase catalytic subunit in normal human cells can lead to bypass of normal senescence, and that inactivation of p16 gene is associated with the process of immortalization. Our present study focuses on assessing telomerase activation and methylation status in the pi6 gene promoter in a panel of human bronchial epithelial (HBE) cells in relation to smoking and previous cancer history. HBE cells were obtained from airway brushes performed during fluorescent bronchoscopy, from the bronchial margin of lung lobes removed during surgery for lung cancer, or from discarded native lungs at the time of lung transplantation. of a panel of 30 HBE cultures, 19 have been analyzed for telomerase activation and ~16 methylation status: 12 from patients with-, and 7 from individuals without lung cancer. The group consisted of 9 smokers, 5 ex-smokers, and 5 non-smokers. Telomerase activity was determined using the telomeric repeat amplification protocol (TRAP), using fluorescent oligonucleotides and capillary electrophoresis detection. Methylation status in the promoter region of p16 was evaluated using methylation specific polymerase chain reaction (PCR) after bisulfite modification. No telomerase activity was detectable in HBE cells of 5 lifetime nonsmokers and in 2 smokers with no lung cancer. Telomerase activity was de-