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Expression of glutathione S-transferase π gene in human lung cancer cell lines
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Expression of Glutathione S-Transferase x Gene in Human Lung Cancer Cell Lines K. Nakagawa, M. Fukuoka, N. Masuda, M. Takada, S. Kudoh, Y. Kusunoki, S. Kishimoto, Department of internal Medicine, Osaka Prefectural Habikino Hospital Resistance to anticancer agents is one of the most important problems in cancer chemotherapy. Glutathione S-transferase (GST) has recently been reported to be one of the factors of resistance to several anticancer agents. To know whether GST-K, which is the most popular isozyme of GST, is responsible for drug resistance, we examined the levels of GST-x mRNA in human lung cancer cell lines and compared with their clinical response to cancer chemotherapy. The 24 lung cancer cell lines used in this study were 3 SCLC, 18 adenocarcinoma, and 3 large cell carcinoma cell lines. Total RNA was prepared by acid guanidinium thiocyanatephenol-chloroform extraction method reported by Piotr Chomczynski et al. The levels of GST-7c mRNA were estimated by RT-PCR using their levels of f32microclobulin (02 MG) mRNA as an internal standard. The results by RT-PCR were analyzed by densitometer and expressed as GST-rr index(GST-x AU’mml02 MG AU’mm). Average GST-x index of 21 NSCLC and 3 SCLC cell lines were 4.5 and 7.9 respectively. The 8 patients in NSCLC were received with MVP regimen and 7 of 8 patients were evaluable. Average GST-x index of Non-responder (N=4) and responder (N=3) were 7.0 and 1.4, suggesting that high levels of GST-n: mRNA might be a marker of resistance to chemotherapy in NSCLC.
Value of Cytological Assessment in Determining Ability to Obtain a Karyotype in Non-Small Cell Robyn Lukeis, Christina Rudduck, Lung Cancer(NSCLC). Lorna Baird*, Louis Irving* and Margaret Garson. St.Vincent's Dept. of Cytogenetics, Hospital, Repatriation General Hospital, Melbourne, Vic..Aust. To better understand factors contributing to lack of success in obtaining an abnormal karyotype in our we examined cytospin study of primary NSCLC's, preparations made from cell suspensions set up in culture for subsequent chromosome harvest. Tumour samples selected macroscopically to be non-necrotic mechanically disaggregated. Papanicolaou were stained slides were assessed for purity and quality of the tumour cell population and evidence of normal epithelial or haemopoietic cells. Of the 30 samples 14 had assessed, 13 had an abnormal karyotype(A), normal(N) and/or unanalysable(U) metaphases, and 3 had no mitoses(NM). Cytological assessment revealed and 10/14(71%)N,U 3/3(100%)NM had either degenerative tumour cells, few tumour cells and/or a cells. While high proportion of haemopoietic 3/13(23%)A had evidence of some tumour degradation, all others had relatively pure, healthy tumour cell population in some cases with haemopoietic cells 13 samples were shown to have present. Therefore, inadequate quality of tumour cells, while of the 17 samples with cytological evidence of good quality tumour cells, 13(76%) produced abnormal karytoypes of sufficient quality. Normal metaphases probably represent dividing normal cells shown to be present. proportion of A higher sq"amous(9/13) than adenocarcinoma(3/9) or large ce11(0/2) had poor quality tumour cell populations and hence a lower cytogenetic success rate.
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Involved Chromosome Regions Specific Nonrandomly in Cytogenetic Abnormalities in Primary Non-Small Cell Lung Cancer(NSCLC). Robyn Garson. Dept. Of Lukeis, Lo" Irving and Margaret St.Vincent's Hospital, Repatriation Cytogenetics, General Hosptial, Melbourne, Vie, Aust. In a study of 18 additional primary NSCLC's we observations of original expand confirm and abnormalities in primary chromosome nonrandom The total series of 28 cases now NSCLC(ref.). includes 9 squamous cell, 13 adenocarcinoma and 6 large cell carcinomas. Despite the complex nature of chromosome distinct karyotypes in most cases, regions are involved in structural abnormalities. The region most frequently involved was 9q13lpll-P22(19/28), followed by p22(21/28), 3qll19pll-P13(14/28), 13,14&/orlSp(15/28), 5qllp23(12/28), 6qll-q15(12/28), 11q11-p15(12/28), lqll-q21(10/28) and Sqll-p12(10/28). q13(11/28), include abnormalities numerical Nonrandom -21(11/28), -16(11/28), +7(11/28), -4(10/28) and -Y overall estimated tumour ploidy. relative to Conclusions from this data confirm loss of 9p as being a frequent finding in primary NSCLC(lS/lS). Rearrangement of lp, lq, lip, 19p, and loss of 3P,8P, 17Pr Sqll-q13 and 6q are also highlighted as consequences of nonrandom chromosome abnormalities. These findings pinpoint regions in which genes whose loss or rearrangement are possibly significant to NSCLC turnour development. Ref.: Genes Chromosomes and Cancer 2:116-124(1990).
CO-AMPLIFICATION OF PARATHYROID HORnONE-RELATED PROTEIN (PTHrP) AND A HUbtAN C-AI-MS-2 IN LUNG CANCER CELL LINE. Christina Rodduck', Lisa Duncanl, Robert o.mrgaret centar2, Garson2. Departments of Medicine1 and Cytogenetics2, St. Vincent's Hospital, Melbourne, Victoria, Australia. Parathyroid hormone-related protein (PTHrP) is a product turnour and the mediator of parathyroid hormone (PTH)-like symptoms of humoral hypercalcaemia of malignancy (HHM). We report on studies of gene amplification in a human lung cancer cell line which overexpresses PTHrP. A large homogeneously staining region (hsr) was found on a marker chromosome and by in situ hybridization we identified this as an area of amplified PTHrP. Using a DNA probe for c-Kiras-2, amplification of this oncogene was also found in the same hsr indicating co-amplification of the c-Ki-ras-2 gene and the gene for PTHrP. These two genes have both been localized to the short arm of chromosome 12. The marker chromosome was dicentric and both centromeres originated from chromosome 16 as demonstrated by in situ hybridization with a centromeric probe for this chromosome. These results are significant as hypercalcaemia is a c0mm0* complication of several C~~C~~.T and occurs frequently in patients with metastatic deposits to the bone.
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Expression of Glutathione S-Transferase x Gene in Human Lung Cancer Cell Lines K. Nakagawa, M. Fukuoka, N. Masuda, M. Takada, S. Kudoh, Y. Kusunoki, S. Kishimoto, Department of internal Medicine, Osaka Prefectural Habikino Hospital Resistance to anticancer agents is one of the most important problems in cancer chemotherapy. Glutathione S-transferase (GST) has recently been reported to be one of the factors of resistance to several anticancer agents. To know whether GST-K, which is the most popular isozyme of GST, is responsible for drug resistance, we examined the levels of GST-x mRNA in human lung cancer cell lines and compared with their clinical response to cancer chemotherapy. The 24 lung cancer cell lines used in this study were 3 SCLC, 18 adenocarcinoma, and 3 large cell carcinoma cell lines. Total RNA was prepared by acid guanidinium thiocyanatephenol-chloroform extraction method reported by Piotr Chomczynski et al. The levels of GST-7c mRNA were estimated by RT-PCR using their levels of f32microclobulin (02 MG) mRNA as an internal standard. The results by RT-PCR were analyzed by densitometer and expressed as GST-rr index(GST-x AU’mml02 MG AU’mm). Average GST-x index of 21 NSCLC and 3 SCLC cell lines were 4.5 and 7.9 respectively. The 8 patients in NSCLC were received with MVP regimen and 7 of 8 patients were evaluable. Average GST-x index of Non-responder (N=4) and responder (N=3) were 7.0 and 1.4, suggesting that high levels of GST-n: mRNA might be a marker of resistance to chemotherapy in NSCLC.
Value of Cytological Assessment in Determining Ability to Obtain a Karyotype in Non-Small Cell Robyn Lukeis, Christina Rudduck, Lung Cancer(NSCLC). Lorna Baird*, Louis Irving* and Margaret Garson. St.Vincent's Dept. of Cytogenetics, Hospital, Repatriation General Hospital, Melbourne, Vic..Aust. To better understand factors contributing to lack of success in obtaining an abnormal karyotype in our we examined cytospin study of primary NSCLC's, preparations made from cell suspensions set up in culture for subsequent chromosome harvest. Tumour samples selected macroscopically to be non-necrotic mechanically disaggregated. Papanicolaou were stained slides were assessed for purity and quality of the tumour cell population and evidence of normal epithelial or haemopoietic cells. Of the 30 samples 14 had assessed, 13 had an abnormal karyotype(A), normal(N) and/or unanalysable(U) metaphases, and 3 had no mitoses(NM). Cytological assessment revealed and 10/14(71%)N,U 3/3(100%)NM had either degenerative tumour cells, few tumour cells and/or a cells. While high proportion of haemopoietic 3/13(23%)A had evidence of some tumour degradation, all others had relatively pure, healthy tumour cell population in some cases with haemopoietic cells 13 samples were shown to have present. Therefore, inadequate quality of tumour cells, while of the 17 samples with cytological evidence of good quality tumour cells, 13(76%) produced abnormal karytoypes of sufficient quality. Normal metaphases probably represent dividing normal cells shown to be present. proportion of A higher sq"amous(9/13) than adenocarcinoma(3/9) or large ce11(0/2) had poor quality tumour cell populations and hence a lower cytogenetic success rate.
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122
Involved Chromosome Regions Specific Nonrandomly in Cytogenetic Abnormalities in Primary Non-Small Cell Lung Cancer(NSCLC). Robyn Garson. Dept. Of Lukeis, Lo" Irving and Margaret St.Vincent's Hospital, Repatriation Cytogenetics, General Hosptial, Melbourne, Vie, Aust. In a study of 18 additional primary NSCLC's we observations of original expand confirm and abnormalities in primary chromosome nonrandom The total series of 28 cases now NSCLC(ref.). includes 9 squamous cell, 13 adenocarcinoma and 6 large cell carcinomas. Despite the complex nature of chromosome distinct karyotypes in most cases, regions are involved in structural abnormalities. The region most frequently involved was 9q13lpll-P22(19/28), followed by p22(21/28), 3qll19pll-P13(14/28), 13,14&/orlSp(15/28), 5qllp23(12/28), 6qll-q15(12/28), 11q11-p15(12/28), lqll-q21(10/28) and Sqll-p12(10/28). q13(11/28), include abnormalities numerical Nonrandom -21(11/28), -16(11/28), +7(11/28), -4(10/28) and -Y overall estimated tumour ploidy. relative to Conclusions from this data confirm loss of 9p as being a frequent finding in primary NSCLC(lS/lS). Rearrangement of lp, lq, lip, 19p, and loss of 3P,8P, 17Pr Sqll-q13 and 6q are also highlighted as consequences of nonrandom chromosome abnormalities. These findings pinpoint regions in which genes whose loss or rearrangement are possibly significant to NSCLC turnour development. Ref.: Genes Chromosomes and Cancer 2:116-124(1990).
CO-AMPLIFICATION OF PARATHYROID HORnONE-RELATED PROTEIN (PTHrP) AND A HUbtAN C-AI-MS-2 IN LUNG CANCER CELL LINE. Christina Rodduck', Lisa Duncanl, Robert o.mrgaret centar2, Garson2. Departments of Medicine1 and Cytogenetics2, St. Vincent's Hospital, Melbourne, Victoria, Australia. Parathyroid hormone-related protein (PTHrP) is a product turnour and the mediator of parathyroid hormone (PTH)-like symptoms of humoral hypercalcaemia of malignancy (HHM). We report on studies of gene amplification in a human lung cancer cell line which overexpresses PTHrP. A large homogeneously staining region (hsr) was found on a marker chromosome and by in situ hybridization we identified this as an area of amplified PTHrP. Using a DNA probe for c-Kiras-2, amplification of this oncogene was also found in the same hsr indicating co-amplification of the c-Ki-ras-2 gene and the gene for PTHrP. These two genes have both been localized to the short arm of chromosome 12. The marker chromosome was dicentric and both centromeres originated from chromosome 16 as demonstrated by in situ hybridization with a centromeric probe for this chromosome. These results are significant as hypercalcaemia is a c0mm0* complication of several C~~C~~.T and occurs frequently in patients with metastatic deposits to the bone.