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Adriamycin resistance in human lung cancer cell lines
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sensitivity. In the 26 instances, association between in vitro drug sensitivity and clinical response to chemotherapy was observed. Of the 26 instances, 24 (92%) showed a correlation between in vitro sensitivity and clinical response. A true positive rate was 50% and a true negative rate 100%. In summary, the results indicated that the tumor stem cell assay was an excellent technique for testing in vitro sensitivity of anticancer drugs. In Vitro Chem0sensitivity Testing For Human Non-Small Cell Lung Cancer: The Role of Cell Lines. Ruckdeschel, J.C., Gazdar, A.F., Carney, D. N., Russel, E.F., Oie, H. NCI-Navy Medical Oncology Branch, Naval Medical Center, Bethesda, MD, United States. Eleven non-small cell lung cancer (NSCLC) cell lines were established from previously untreated patients (8 continuous, 3 recently cultured) and tested for responsiveness to doxorubicin (D), mitomycin (M), cisplatin (P), and etoposide (E) using a non-clonogenic dye-exclusion assay originally developed by Weisenthal (Ca. Res. 43: 749, 1983). In vitro drug doses were determined empirically (D, 1.2; M, 0.8; P, 6.6; VP 125; all ug/ml) and tested across a 3 log dose range with a 1 hr exposure. These doses more closely approximate the CXT concentrations for these drugs. Total refractoriness was seen to D 40% of the time and to M and P 53% of the time. All cell lines tested were sensitive to the highest dose of E and 33% were sensitive to the "standard" dose. Thirteen percent of tests for D and P and 7% for M were sensitive (< 50% cell survival) at the "standard dose". Only 4 of the continuous cell lines cloned sufficiently in agarose to permit drug testing by the traditional clonogenic assay. Concurrent clonogenic and non-clonogenic assays for 3 of these lines showed a similar pattern of drug activity but different absolute values for % cell survival. This technique permits more rapid evaluation of results (4d vs 2-3w), can be performed on virtually all cell lines, demonstrates consistent dose response curves and shows va~-iation between lines with respect to drug sensitivity. The use of continuous cell lines and a non-clonogenic assay should permit in vitro, phase II testing specific to a particular malignancy. Use of the assay with recently established cultures is currently being tested for individual drug testing.
Adriamycin Resistance in Human Lung Cancer Cell Lines. Twentyman, P.R., Fox, N.E., Wright, K.A., Bleehen, N.M. MRC Clinical Oncology and
Radiotherapeutics Unit, MRC Centre, Cambridge, U.K. We are interested in whether adriamycin (ADM) resistance in human lung cancer cell lines is similar to the pleiotrop~c drug resistance seen in rodent cells (Ling et al, Cancer Treat. Rep., 67, 869, 1983). We have derived ADM resistant sublines of human lung cancer cell lines: NCI-H69 (small cell), MOR (adenocarcinoma) and COR-L23 (large cell anaplastic). The lines were produced by continuous exposure to ADM in the medium followed by a period of drug-free growth and then a return to drug-containing medium at successively increasing doses. The time taken for the resistant sublines to be isolated was 6-9 months compared with 4 weeks for mouse tumor cell lines. The extent of resistance is shown in the table. Cell Line Type
NCI-H69 MOR COB-L23
Continuous ADM dose (ug/ml) to reduce cell growth by 80% Control Resistant
Small cell 0.010 Adeno 0.045 Large cell 0.030
0.85 0.14 0.22
The resistant lines each demonstrate reduced ADM accumulation during a lh exposure. The resistant subline of NCI-H69 also shows resistance to vincristine, colchicine, 4'deoxyadriamycin and 4'epiadriamycin but not to melphalan, CCNU, bleomycin or aclacinomycin A. Its resistance to ADM can at least partly be overcome by the use of verapamil (an inhibitor of membrane calcium transport). We are currently investigating differences in cellular pharmacokinetics for a range of anthracyclines between normal and ADM resistant cells together with possible mechanisms of resistance. Development of Multiple Drug-Resistance in Human Small Cell Lung Cancer (SCLC) Cells by Continuous Exposure to Adriamycin (ADM) in Vitro. Ohnoshi, T., Hiraki, S., Miyamoto, H., Kimura, I. Department of Medicine, Okayama University Hospital, Okayama 700, Japan. A human SCLC cell line resistant to multiple drugs was developed by exposure to increasing concentrations of ADM. After 24 passages of the p a r e n t _ ~ l l line (SB~-3) in the presence of ADM (i0: -M to 5 x 10-M),_a population of cells that survived under i0 6M ADM for 1 hour was collected by cloning in soft agar. The cells (SBC-3/ADM) was 30 times more resistant to ADM than SBC-3. Resistance was stable after 6 months following removal of ADM. SBC-3/ ADM was 28-fold resistant to daunomycin and 26-fold to 4'-epi-ADM when measured in a clonogenic assay. Of interest, SBC-3/ADM was 25-fold resistant to vincristine and 19-fold to mitomycin C. Resistance was intermediate for tetrahydropyranol-ADM, aclarubicin and cisplatin, however, SBC-3/ADM was as equally sensitive to
sensitivity. In the 26 instances, association between in vitro drug sensitivity and clinical response to chemotherapy was observed. Of the 26 instances, 24 (92%) showed a correlation between in vitro sensitivity and clinical response. A true positive rate was 50% and a true negative rate 100%. In summary, the results indicated that the tumor stem cell assay was an excellent technique for testing in vitro sensitivity of anticancer drugs. In Vitro Chem0sensitivity Testing For Human Non-Small Cell Lung Cancer: The Role of Cell Lines. Ruckdeschel, J.C., Gazdar, A.F., Carney, D. N., Russel, E.F., Oie, H. NCI-Navy Medical Oncology Branch, Naval Medical Center, Bethesda, MD, United States. Eleven non-small cell lung cancer (NSCLC) cell lines were established from previously untreated patients (8 continuous, 3 recently cultured) and tested for responsiveness to doxorubicin (D), mitomycin (M), cisplatin (P), and etoposide (E) using a non-clonogenic dye-exclusion assay originally developed by Weisenthal (Ca. Res. 43: 749, 1983). In vitro drug doses were determined empirically (D, 1.2; M, 0.8; P, 6.6; VP 125; all ug/ml) and tested across a 3 log dose range with a 1 hr exposure. These doses more closely approximate the CXT concentrations for these drugs. Total refractoriness was seen to D 40% of the time and to M and P 53% of the time. All cell lines tested were sensitive to the highest dose of E and 33% were sensitive to the "standard" dose. Thirteen percent of tests for D and P and 7% for M were sensitive (< 50% cell survival) at the "standard dose". Only 4 of the continuous cell lines cloned sufficiently in agarose to permit drug testing by the traditional clonogenic assay. Concurrent clonogenic and non-clonogenic assays for 3 of these lines showed a similar pattern of drug activity but different absolute values for % cell survival. This technique permits more rapid evaluation of results (4d vs 2-3w), can be performed on virtually all cell lines, demonstrates consistent dose response curves and shows va~-iation between lines with respect to drug sensitivity. The use of continuous cell lines and a non-clonogenic assay should permit in vitro, phase II testing specific to a particular malignancy. Use of the assay with recently established cultures is currently being tested for individual drug testing.
Adriamycin Resistance in Human Lung Cancer Cell Lines. Twentyman, P.R., Fox, N.E., Wright, K.A., Bleehen, N.M. MRC Clinical Oncology and
Radiotherapeutics Unit, MRC Centre, Cambridge, U.K. We are interested in whether adriamycin (ADM) resistance in human lung cancer cell lines is similar to the pleiotrop~c drug resistance seen in rodent cells (Ling et al, Cancer Treat. Rep., 67, 869, 1983). We have derived ADM resistant sublines of human lung cancer cell lines: NCI-H69 (small cell), MOR (adenocarcinoma) and COR-L23 (large cell anaplastic). The lines were produced by continuous exposure to ADM in the medium followed by a period of drug-free growth and then a return to drug-containing medium at successively increasing doses. The time taken for the resistant sublines to be isolated was 6-9 months compared with 4 weeks for mouse tumor cell lines. The extent of resistance is shown in the table. Cell Line Type
NCI-H69 MOR COB-L23
Continuous ADM dose (ug/ml) to reduce cell growth by 80% Control Resistant
Small cell 0.010 Adeno 0.045 Large cell 0.030
0.85 0.14 0.22
The resistant lines each demonstrate reduced ADM accumulation during a lh exposure. The resistant subline of NCI-H69 also shows resistance to vincristine, colchicine, 4'deoxyadriamycin and 4'epiadriamycin but not to melphalan, CCNU, bleomycin or aclacinomycin A. Its resistance to ADM can at least partly be overcome by the use of verapamil (an inhibitor of membrane calcium transport). We are currently investigating differences in cellular pharmacokinetics for a range of anthracyclines between normal and ADM resistant cells together with possible mechanisms of resistance. Development of Multiple Drug-Resistance in Human Small Cell Lung Cancer (SCLC) Cells by Continuous Exposure to Adriamycin (ADM) in Vitro. Ohnoshi, T., Hiraki, S., Miyamoto, H., Kimura, I. Department of Medicine, Okayama University Hospital, Okayama 700, Japan. A human SCLC cell line resistant to multiple drugs was developed by exposure to increasing concentrations of ADM. After 24 passages of the p a r e n t _ ~ l l line (SB~-3) in the presence of ADM (i0: -M to 5 x 10-M),_a population of cells that survived under i0 6M ADM for 1 hour was collected by cloning in soft agar. The cells (SBC-3/ADM) was 30 times more resistant to ADM than SBC-3. Resistance was stable after 6 months following removal of ADM. SBC-3/ ADM was 28-fold resistant to daunomycin and 26-fold to 4'-epi-ADM when measured in a clonogenic assay. Of interest, SBC-3/ADM was 25-fold resistant to vincristine and 19-fold to mitomycin C. Resistance was intermediate for tetrahydropyranol-ADM, aclarubicin and cisplatin, however, SBC-3/ADM was as equally sensitive to