of 1 /1
33 125 IDENllFlCA~ OF TYFB-BFECiFlC GENETIC BIOMARKERS FOB LUNG CANCER IJBfNG UFFBBBNTtAL mRNA EXPRESSION RNQmlPmNnm I. Newshamll2, D. Daub’ , V. Russack3 J. Harrell2, H. Co@, J. Green4, W. Cavenee1p2, and M. Green2. i Ludwfg Institute for Cancer Research-San Diego Branch, Depts. of 2Medldne and 3Pathology. and 4UCSD Cancer Center, UCSD School of Medfcine, La Jolla, CA, USA. examinod in more de biomarkers for lun cancer itself and in areas of premalignant change in bronchial epithe turn. P PRIMARY NORMAL HUMAN BRONCHIAL EPITHELIAL (NHBE) CELLS AS A MODEL SYSTEM FOR CYPIAI INDUCTION IN LUNG CARCINOGENESISAND XENO-BIOTIC METABOLISM J.E. Jones, SM. Jensen, L.E. Beebe’ , L.M. Anderson’ , R.I. Linnoila Biomarkers & Prevention Research Branch, NCI, NIH, Rockville, MD 20850 and ‘Laboratory of Comparative Carcinogenesis, NCI, NlH, Frederick, MD 21207. Elevated levels of expression of the CYPlAI gene have been associated with an increased risk of lung cancer and correlate with increased mortality. In previous studies on well characterized human non-small cell lung cancer derived cell lies we have identified a unique pattern ofregulation ofthe CYPIAI gene. To further elucidate the responses of specific, non-neoplastic populations of target cells within the lung to exposure to chemical agents we have used commercially available NHBE cell “lines”. Each primary cell culture was established from a single “donor” at autopsy. Cells were routinely propagated for two to four passages in mod&d LHC-9 medium containing 0.4% bovine pituitary extract prior to analysis. Characterization by immunobistochemistry and in situ RNA-RNA hybridization demonstrated that, while most NHBE cells were cytokeratin positive, sub-populations composed of variable numbers of basal cells, non-ciliated secretory cells and endocrine cells positive for keratin 14, CC10 and chmmogranin, respectively, were present. The levels of expression of the CYPlAl gene, under control and inducing conditions, were compared to EROD activities and the patterns are discussed. We conclude that NHBE cells may vary in marker and enzyme expression, but provide a useful tool in studying the contribution of each cell “type” to the total expression of CYPlAI activity in various cell types innormalhuman airways. 126 DETECTION OF HUBAN PAPILLOMAVIRUS DNA AND E6-E7 mRNA IN LUNG CANCER. I. Xinoshita, H. Dosaka-Akita, M. Fujino, K. Akie, M. Kato, Y. Kawakami. First Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan. In lung cancer, HPV DNA has been detected in several percent of squamous cell carcinomas using in situ DNA hybridization and Southern blot hybridization. To provide an accurate evaluation of the association of human papillomavirus (HPV) with lung cancer, 36 cases of lung cancer were analyzed for HPV DNA by the extremely sensitive polymerase chain reaction (PCR) method and dot blot hybridization, and for the transcription from the E6-E7 transforming region by in situ hybridization. HPV-18 DNA was detected in three (8%) of 36 specimens; histologically, in one (10%) of 10 squamous cell carcinomas and two (9%) of 22 adenocarcinomas. Neither HPV-16 nor -33 DNA was detected in any cases examined. Expression of Eb- E7 mRNA was observed in a case of squamous Cell carcinoma and a case of adenocarcinoma which contained HPV-IS DNA. All HPV-positive cases were male smokers. Pathologically, the case of squamous cell carcinoma was moderately differentiated without koilocytotic feature. One HPV-positive adenocarcinoma was moderately differentiated acinar adenocarcinoma in which cytoplasmic mucin and mitosis were often observed. The other was well differentiated papillary adenocarcinoma. HPV-18 may play an important role in the development and progression of cancer in some cases of both squamous cell carcinoma and adenocarcinoma of the luna. 12s yATg;gTIAL HYPERTENSION IN LUNG CARCINOMAS IN Y. Boucher,‘M. Nozue, J. Wain, D. Math&n, H.C. Grille, N. Choi, and R.K. Jam. Dept. of Radiation Oncology and Thoracic Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114. The penetration in solid tumors of oxygen, glucose and low molecular weight blood bome therapeutic agents is determined by the functional vascular surface area and blood perfusion. Furthermore, for larger molecules (e.g. monoclonal antibodies), extravasation could be significantly impeded by tbe structure of the. micmvascular wall and the negligible hydrostatic pressure gradient between the microvascular and interstitial space. Recent studies have demonstrated that elevated interstitial fluid uressure (IFP) is a oatboubvsioloaical characteristic of human tools in Situ (melanomas a&d ca&tomas~of the cervix, breast, colorectal. kidnev and head and neck). In this sNdv we mesent data on the lFP in the 4 major hlstologlcal types of prhnaj lung Carcinomas. IFP was measured in 24 patients wltb stage I or II lung cancer. Under general anasthesia, prior to lobectomy or pneumonectomy, two IFP measurements were made uer Nmor with tbe wick - in - needle technology (Bomber et al., dancer Res, 1991). In 504b of the tumors the difference between the 2 IFP measurements in a tumor was less than 25%. whereas in the other Nmors it varied between 25 and 100%. In adenocarcinomas (n=13) IFP varied between 1.0 and 27.0 mm Hg with a mean of 8.0 f 4.0 (SD). IFP in sqamous cell carcinomas (n=9) varied between 4.0 and 27 mm Hg with a mean of 14.0 f 8.0 mm Hg. The pressures in one small cell carcinoma and one large cell carcinoma were respectively, 15 and 14 mm Hg. In tbe normal lung (n = 4). IFP varied between -1.0 and 2.0 mm Hg with a mean IFP of 0.0 mm Hg. In conclusion the measurements demonstrate that interstitial hypertension is also a cbaracteristlc of lung carcinomas. Further measurements arc being made N determine if there is a significsnt difference in pressure among the different histological types of lung tumors. [Supported by NC1 - CA - 565911.

Primary normal human bronchial epithelial (NHBE) cells as a model system for CYP1A1 induction in lung carcinogenesis and xeno-biotic metabolism

Embed Size (px)

Citation preview

33

125

IDENllFlCA~ OF TYFB-BFECiFlC GENETIC BIOMARKERS FOB LUNG CANCER IJBfNG UFFBBBNTtAL mRNA EXPRESSION RNQmlPmNnm I. Newshamll2, D. Daub’, V. Russack3 J. Harrell2, H. Co@, J. Green4, W. Cavenee1p2, and M. Green2. i Ludwfg Institute for Cancer Research-San Diego Branch, Depts. of 2Medldne and 3Pathology. and 4UCSD Cancer Center, UCSD School of Medfcine, La Jolla, CA, USA.

examinod in more de

biomarkers for lun cancer itself and in areas of premalignant change in bronchial epithe turn. P

PRIMARY NORMAL HUMAN BRONCHIAL EPITHELIAL (NHBE) CELLS AS A MODEL SYSTEM FOR CYPIAI INDUCTION IN LUNG CARCINOGENESISAND XENO-BIOTIC METABOLISM J.E. Jones, SM. Jensen, L.E. Beebe’, L.M. Anderson’, R.I. Linnoila Biomarkers & Prevention Research Branch, NCI, NIH, Rockville, MD 20850 and ‘Laboratory of Comparative Carcinogenesis, NCI, NlH, Frederick, MD 21207.

Elevated levels of expression of the CYPlAI gene have been associated with an increased risk of lung cancer and correlate with increased mortality. In previous studies on well characterized human non-small cell lung cancer derived cell lies we have identified a unique pattern of regulation of the CYPIAI gene. To further elucidate the responses of specific, non-neoplastic populations of target cells within the lung to exposure to chemical agents we have used commercially available NHBE cell “lines”. Each primary cell culture was established from a single “donor” at autopsy. Cells were routinely propagated for two to four passages in mod&d LHC-9 medium containing 0.4% bovine pituitary extract prior to analysis. Characterization by immunobistochemistry and in situ RNA-RNA hybridization demonstrated that, while most NHBE cells were cytokeratin positive, sub-populations composed of variable numbers of basal cells, non-ciliated secretory cells and endocrine cells positive for keratin 14, CC10 and chmmogranin, respectively, were present. The levels of expression of the CYPlAl gene, under control and inducing conditions, were compared to EROD activities and the patterns are discussed. We conclude that NHBE cells may vary in marker and enzyme expression, but provide a useful tool in studying the contribution of each cell “type” to the total expression of CYPlAI activity in various cell types in normal human airways.

126

DETECTION OF HUBAN PAPILLOMAVIRUS DNA AND E6-E7 mRNA IN LUNG CANCER. I. Xinoshita, H. Dosaka-Akita, M. Fujino, K. Akie, M. Kato, Y. Kawakami. First Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan.

In lung cancer, HPV DNA has been detected in several percent of squamous cell carcinomas using in situ DNA hybridization and Southern blot hybridization.

To provide an accurate evaluation of the association of human papillomavirus (HPV) with lung cancer, 36 cases of lung cancer were analyzed for HPV DNA by the extremely sensitive polymerase chain reaction (PCR) method and dot blot hybridization, and for the transcription from the E6-E7 transforming region by in situ hybridization.

HPV-18 DNA was detected in three (8%) of 36 specimens; histologically, in one (10%) of 10 squamous cell carcinomas and two (9%) of 22 adenocarcinomas. Neither HPV-16 nor -33 DNA was detected in any cases examined. Expression of Eb- E7 mRNA was observed in a case of squamous Cell carcinoma and a case of adenocarcinoma which contained HPV-IS DNA. All HPV-positive cases were male smokers. Pathologically, the case of squamous cell carcinoma was moderately differentiated without koilocytotic feature. One HPV-positive adenocarcinoma was moderately differentiated acinar adenocarcinoma in which cytoplasmic mucin and mitosis were often observed. The other was well differentiated papillary adenocarcinoma.

HPV-18 may play an important role in the development and progression of cancer in some cases of both squamous cell carcinoma and adenocarcinoma of the luna.

12s

yATg;gTIAL HYPERTENSION IN LUNG CARCINOMAS IN

Y. Boucher,‘M. Nozue, J. Wain, D. Math&n, H.C. Grille, N. Choi, and R.K. Jam. Dept. of Radiation Oncology and Thoracic Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.

The penetration in solid tumors of oxygen, glucose and low molecular weight blood bome therapeutic agents is determined by the functional vascular surface area and blood perfusion. Furthermore, for larger molecules (e.g. monoclonal antibodies), extravasation could be significantly impeded by tbe structure of the. micmvascular wall and the negligible hydrostatic pressure gradient between the microvascular and interstitial space. Recent studies have demonstrated that elevated interstitial fluid uressure (IFP) is a oatboubvsioloaical characteristic of human tools in Situ (melanomas a&d ca&tomas~of the cervix, breast, colorectal. kidnev and head and neck). In this sNdv we mesent data on the lFP in the 4 major hlstologlcal types of prhnaj lung Carcinomas. IFP was measured in 24 patients wltb stage I or II lung cancer. Under general anasthesia, prior to lobectomy or pneumonectomy, two IFP measurements were made uer Nmor with tbe wick - in - needle technology (Bomber et al., dancer Res, 1991). In 504b of the tumors the difference between the 2 IFP measurements in a tumor was less than 25%. whereas in the other Nmors it varied between 25 and 100%. In adenocarcinomas (n=13) IFP varied between 1.0 and 27.0 mm Hg with a mean of 8.0 f 4.0 (SD). IFP in sqamous cell carcinomas (n=9) varied between 4.0 and 27 mm Hg with a mean of 14.0 f 8.0 mm Hg. The pressures in one small cell carcinoma and one large cell carcinoma were respectively, 15 and 14 mm Hg. In tbe normal lung (n = 4). IFP varied between -1.0 and 2.0 mm Hg with a mean IFP of 0.0 mm Hg. In conclusion the measurements demonstrate that interstitial hypertension is also a cbaracteristlc of lung carcinomas. Further measurements arc being made N determine if there is a significsnt difference in pressure among the different histological types of lung tumors. [Supported by NC1 - CA - 565911.