The effect of high intensity focused ultrasound on luciferase activity on two tumor cell lines in vitro, under the control of a CMV promoter.
In this study, we compared the effect of high intensity focused ultrasound (HIFU) and thermal stress on the luciferase activity, controlled by a cytomegaly virus (CMV) promoter in an in vitro model using two tumor cell lines (M21, SCCVII). HIFU was applied in a pulsed-wave mode with increasing voltage at constant pulse duration, or thermal stress was delivered over a range of temperatures (36-52 degrees C) for 5 min. The resulting luciferase activity was measured in live cells using a cooled CCD camera. Luciferase activity was measured at set time intervals over a total of 48 h post-stress. Compared to baseline, the luciferase activity of the M21 tumor cell line when exposed to HIFU was approximately 54.2+/-67.5% (p<0.01) higher at a temperature of 42 degrees C, and approximately 52.9+/-128.5% (p<0.01) higher at 44 degrees C. In the SCCVII tumor cell line, the luciferase activity after HIFU application was 55.4+/-66.6% (p<0.01) higher compared to baseline at a temperature of 42 degrees C. The M21 and SCCVII tumor cell line when exposed to thermal stress alone did not increase the luciferase activity. M21 and SCCVII tumor cells exposed to HIFU showed a maximum decrease in cell viability to 45.3+/-7.5% and 10.3+/-7.5%, respectively, and when exposed to thermal stress to 85.3+/-3.5% and 20.4+/-6.5%, respectively, compared to the untreated control. In M21 and SCCVII cells exposed to HIFU, free radicals could be detected using the dichlorofluorescein dye. Our findings demonstrate that HIFU can enhance the luciferase activity controlled by a CMV promoter. However it also has a higher damaging effect on the cells.
Hundt
Walter
W
Department of Radiology, Lucas MRS Research Center, Stanford School of Medicine, Stanford, CA 94305, USA. walter.hundt@web.de
Steinbach
Silke
S
O'Connell-Rodwell
Caitlin E
CE
Bednarski
Mark D
MD
Guccione
Samira
S
D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000503:physiopathology*; D045744:Cell Line, Tumor; D003587:Cytomegalovirus / Q000235:genetics*; D004789:Enzyme Activation / Q000528:radiation effects; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism*; D008545:Melanoma / Q000503:physiopathology*; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics; D013010:Sonication; D014162:Transfection / Q000379:methods*
D008156:Luciferases