Assessments of the Sensitivities of Cultured Human Neuroblastoma Cells to Anti-Tumour Drugs

Abstract

SummaryWhen logarithmically growing neuroblastoma cells were exposed in vitro for 24 hr to a range of concentrations of various antitumour drugs and then cloned in agar to assess colony-forming ability, the dose-response curves obtained were of two types only, namely exponential-plateau or exponential. The former category, included hydroxyurea, 1.1'-propylene bispiperazine-3.5-dione (ICRF-159). vincristine and vindesine. where the plateau was reached before 1 log cell kill, and 4'-demethylepipodophllotoxin thenylidine glucoside (NSC 122819) (VM-26) and 4'-demethyle p i p o d o p h l l o t o x i n -9 -( 4 , 6 -o e t h y l i d i n e -~y r a n o d e (NSC 141540) . where the plateau occurred at approximately 5% survival. Exponential survival curves were obtained with actinomycin D, Adriamycin, bleomycin, cis-dichlorodiammine platinum (11) (cis-platinum), dihromodulcitol, Ifluorouracil, 4'-(9-acridinylamino)methanesuIfon-m-anisidine (NSC 249992) (mAMSA), melphalan, and peptichemio. A comparison of drug concentrations required to reduce survival in vitro by 70%. with a 2 4 h r drug exposure, was attempted with those concentrations achievable as plasma levels in man after conventional drug therapy. The results obtained, after many and varied assumptions, allowed the division of the drugs tested into three main groups: (1) most effective agents, including mAMSA, VM-26, and Adriamycin; (2) agents with some activity-vincristine, vindesine, VP-16-213, cis-platinum, melphalan, and peptichemio, although only when the latter three drugs were used at the higher dosage levels; (3) agents with little, if any, activity-actinomycin D. bleomycin, dibromodulcitol, 5-fluorouracil, hydroxyurea, and ICRF-159. In terms of attempting clinical correlations, available data suggest definite value in treat-, ing neurohlastoma with VM-26 and Adriamycin [from our group ( I ) ) and also with vincristine, melphalan (high dose), peptichemio, and cis-platinum [from our group (2)l. No significant clinical activity has been reported for actinomycin D nor for bleomycin in previously treated children, and both these drugs proved negative in this model test system. Among the newer agents tested experimentally, our results suggest that although clinical evaluation of mAMSA, VP-16-213, and vindesine might be profitahle, dibromodulcitol and ICRF-159 may prove inactive. SpeculationThe dismal prognosis for the majority of children with neuroblastoma serves to highlight the need for new and more effective therapies. One approach is to attempt to screen experimentally in vitro for drugs effective in this disease, Our preliminary studies, using only one cell line derived from one portion of a human tumour, have suggested, with many reservations, some correlations between in virro and clinical data. This work is now being extended to consider other human neuroblastoma cell lines and to monitor retrospectively drug responses in vitro with clinical responses. These studies are essential before any attempts are made to extrapolate any of the in ~i t r o results to...

The survival of cytochalasin‐induced polykaryons following exposure to cytotoxic agents.

“…This drug has previously been found to yield biphasic curves, and the initial slope has been suggested to represent the elimination of a sensitive (possibly S) subpopulation (Madoc-Jones and. Mauro,1968;Drewinko and Barlogie,1981;Hill and Whelan, 1981). The 16C+ curve in CHO-Kl approximated to an exponential whose slope was similar to the second clonogenic phase.…”

High dose melphalan in the treatment of advanced neuroblastoma: Results of a randomised trial (ENSG‐1) by the European Neuroblastoma Study Group

“…The log‐linear in vitro dose‐response of cultured neuroblastoma cells to melphalan and other alkylating agents 10 provides a rationale for “high dose consolidation” chemotherapy (“megatherapy”) to try to overcome drug resistance 11. Since its introduction into clinical practice in 1977, high dose melphalan has been used by many centres to treat patients with advanced neuroblastoma, often in combination with either total body irradiation (TBI) or other high dose cytotoxic drugs 12–14.…”

Use of human neuroblastoma continuous cell lines for in vitro drug sensitivity screening

“…It can now be seen, for example, after using the three lines that in our original study [1] the CHP100 cells were hypersensitive to amsacrine, methotrexate and vincristine, yet refractory to cisplatin and VP-16. So if we applied the effectiveness criteria, described earlier [1], on the basis of the three cell lines we would now exclude amsacrine as the most effective agent, but promote VP-16 and 'high-dose' cisplatin to join adriamycin and VM 26 and downgrade the Vinca alkaloids. Such a classification appears to fit more closely with clinical experience, since it has now been shown that amsacrine is ineffective against neuroblastoma [12], whilst VP-16 plus high-dose cisplatin has demonstrated activity in this disease [13].…”

“…The same method described in detail earlier [1] was used for these neuroblastoma cell lines. Briefly, cells were plated at a density of 2 • 104/ml into 2 cm dishes and two days later, a range of concentration of each drug were added in 50/zl volumes to each dish.…”

“…In initial studies [1] we showed that it was possible, using one human neuroblastoma cell line in vitro, to evaluate the effectiveness of a range of antitumor drugs using colonyforming assays and to attempt correlations between the effective lethal drug concentrations in vitro with those achievable clinically. We have now extended these studies using three continuous human neuroblastoma cell lines and have also evaluated certain newer investigational agents.…”