Treatment of artificially-induced pulmonary metastases with fractionated doses of vincristine and/or radiation therapy

Abstract

The cytotoxic effects in vivo of vincristine (VC), radiation, or both modalities in combination on murine fibrosarcoma (FSa) cells grown as pulmonary tumors were determined. Fourteen days following the i.v. injection of viable FSa cells, recipient mice developed between 100 and 150 visible pulmonary nodules. At that time, tumor-bearing animals were exposed to either single or combined modality treatments, as well as single and fractionated dose regimens. Animals were sacrificed 1 hour after the last treatment. Tumor nodules were excised and made into a single cell suspension and separated on the basis of cell size by centrifugal elutriation. Flow microfluorometry (FMF) was used to determine the cell-cycle parameters and the relative synchrony of the separated populations, as well as the percentage contamination by normal diploid cells in each of the tumor cell populations. Known numbers of viable cells from each elutriator fraction were injected into recipient mice to determine their colony-forming efficiency (CFE). Surviving fractions were determined by comparing the CFEs of treated FSa cells from each of the separated elutriator fractions with those of appropriate untreated controls. Following a single dose of VC (1 mg/kg), populations of cells enriched in late S and G2 + M were the most sensitive. However, following the administration of five doses (0.25 mg/kg each) over a 24 hour period, populations of cells most enriched in G1 cells exhibited the lowest percentage of survivors. A single dose of radiation (1000 rad) was most effective in killing S-phase cells. When administered in five fractions (250 rad per fraction), each separated by a 6 hour interval, no phase-specific sensitivity was observed. The combination of both modalities exhibited a marked schedule dependence, with a single dose (1000 rad) of radiation followed by five doses of VC (0.25 mg/kg) being the most effective treatment protocol observed.

Effectiveness of AMSA alone or in combination with radiation on murine fibrosarcoma pulmonary nodules

“…The tumor and cell separation systems have been described in detail elsewhere [12]. Briefly, a methylcholanthrene induced fibrosarcoma (FSa) was used in this study.…”

“…Irradiation was performed using a 137Cs source having a dose rate of 240 rad/min [12]. The drug was administered i.v.…”

“…Following treatment, lungs containing tumor nodules from a minimum of 40 animals per treatment group were excised, made into single cell suspension by mincing and trypsinization, and separated by centrifugal elutriation according to methods described in detail elsewhere [12].…”

“…In a previous communication, the effectiveness of vincristine in combination with fractionated doses of ionizing radiation was described [12]. The combination of both modalities exhibited a marked schedule dependence, with the most effective result occurring when vincristine was administered after irradiation.…”

“…An in vivo procedure has been developed which allows for the rapid testing and evaluation of the phase-specific therapeutic effectiveness of selected cytotoxic agents on artificial pulmonary metastases [11][12][13]. In particular, parameters such as the sequencing and timing of therapeutic agents need to be characterized.…”