Lethal Effects of Radiation and Platinum Analogues on Multicellular Spheroids of HeLa Cells

Hong, Seong Eon

Abstract

Multicellul ar tumor spheroids of HeLa cells have been grown in a static culture system. Samples of spheroids were exposed for 2 h to graded concentration of sis-platinum and its analogue, carboplatin, and then response assayed by survival of clonogenic cells. The purpose of present experiment is to clarify the effectiveness of these platinum compounds and to evaluate intrinsic radiosensitivity of cells using spheroids of HeLa cells as an experimental in vitro model. Variations of the drug sensitivity of monolayers as well as spheroids were also evaluated in cell-survival curves. In cia-platinum concentration-survival cutie, there was a large shoulder extending as far as Cq=3.4 mM, after which there was exponential decrease in survival curve having a Co Value of 1.2 mu in spheroids. While the Co for the spheroids was essentially no significant change, but Cq value was larger than that of monolayers. This suggest that the effect of cis-platinum is greater in the monolayer with actively proliferaing cells than hypoxic one. In the carboplatin concentration-survival curves, the Co value of spheroids was 15.0 mM and the ratio with the Co from monolayer cell (32.5 mM) was 0.46, thus indicating that the spheroids had a greater sensitivity to carboplatin than monolayers. Therefore, the effect of carboplatin is mainly on the deeper layers of spheroids acting as hypoxic cell sensitizer. The enhanced effect was obtained for monolayer cells using combined X-ray and carboplatin treatment 2 hours before irradiation. The result shown in isobologram analysis for the level of surviving fraction at 0.01 indicated that the effect of two agents was truely supra-additive. From this experimental data, carboplatin has excited much recept interest as one of the most promising, since it is almost without nephrotoxicity and causes less gastrointestinal toxicity than cia-platinum. Interaction between carboplatin and radiation might play an important role for more effective local tumor control.