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• W3034984195 abstract "3803 Background: In a previous study we showed that a multivariate linear regression model of gene expression and radiation response (as measured by survival fraction at 2Gy, SF2) predicted cellular radiosensitivity and identified at least one novel protein (RbAp48) mechanistically involved in radiation response. The model predicted that three additional known genes (rgs-19, r5pia and topo1) could be involved in regulating radiation response. In this study, we continue the biological testing of the multivariate linear regression model. Methods: The details of the multivariate linear regression model of gene expression and radioresponse in a 35 cell line database have been published previously (1).RGS-19 (Flag -tag) and R5PIA were transfected into MDA-MB231 and MALME-3M cell lines respectively, using Lipofectamine2000 reagent (Invitrogen). Overexpression of both RGS-19 and R5PIA were confirmed by western blot. Cell survival after 2, 4 6,and 8 Gy radiation were determined by the clonogenic assay. RbAp48 siRNA (100 nM) designed by Dharmacon’s SMARTpool technology or a pool of 4 negative controls siRNA (100 nM) was transfected into HCT116 colon cancer cell lines following the DharmaFECT transfection protocol (Dharmacon). Knockdown of RbAp48 was verified 72 hours after transfection, by Western Blot. Results: The model predicted that overexpression of RGS-19 and R5PIA should lead to radiosensitization while downregulation of RbAp48 should lead to radioresistance. Indeed, overexpression of RGS-19 resulted in radiosensitization of MDA-MB231 breast cancer cell lines (SF2, Empty Vector: 0.50 ± 0.06 vs. RGS19 plasmid: 0.35 ± 0.06, n=3, p=0.01) (SF8 value, Empty Vector: 0.096 ± 0.053 and RGS19 plasmid: 0.068 ± 0.06, n=3, p=0.06). In contrast, overexpression of R5PIA had no effects on radiosensitivity of SKMEL28 cancer cell lines as measured by SF2 (Empty Vector: 0.64 ± 0.10 and R5PIA plasmid: 0.68 ± 0.09, n=4). Additionally downregulation of RbAp48 resulted in radiation resistance of HCT-116 (SF2, control siRNA: 0.26 ± 0.04, RbAp48 siRNA: 0.37 ± 0.03, n=6) (SF8 control siRNA: 0.00275 ± 0.0002, RbAp48 siRNA: 0.00575 ± 0.0004, n=6), also consistent with the predictions of the model. Conclusions: These data further confirm the biological validity of the mathematical algorithm applied to this genomic analysis. Two of the three genes tested affected radiation response as predicted by the model. A third gene (top1) is currently a target for radiosensitization in clinical use. References 1. Torres-Roca JF, Eschrich S, Zhao H, Bloom G, Sung J, McCarthy S, Cantor AB, Scuto A, Li C, Zhang S, Jove R, Yeatman T. (2005) Cancer Res 65:7169-76" @default.
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• W3034984195 date "2007-05-01" @default.
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• W3034984195 title "Biological validation of a linear regression mathematical model of gene expression and radiation response" @default.
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