Cisplatin is one of the most widely used chemotherapeutics in the world today. Unfortunately, chemoresistance often develops hindering the effectiveness of the drug. Mismatch‐repair (MMR) and p53 have previously been shown to be important determinants of cisplatin resistance and can contribute to cisplatin resistance clinically. Here, we have used cDNA microarray to identify several genes as up or downregulated in a previously described, cisplatin resistant, clone of the HCT116 cell line (HCT116‐K). On follow‐up, one gene, APM2, was found to promote cisplatin resistance when overexpressed in sensitive HCT116 clones. Furthermore, silencing APM2 in a panel of cell lines encompassing all combinations of p53 status and MMR proficiency (HCT116‐K, HCT116, SW620, MCF7, PC‐3 and OV2008) resulted in sensitization regardless of these 2 factors. In addition, silencing APM2 stably using shRNA also resulted in the sensitization of cells to cisplatin. More importantly, cisplatin inhibited the growth of APM2 silenced tumor xenografts (HCT116‐K or OV2008 cells) significantly better than it inhibited the growth of xenografts carrying nontargeting control shRNAs. These findings represent a novel strategy that could be exploited to overcome cisplatin resistance in patients regardless of p53 status or ability to perform MMR. © 2009 UICC

Background: As part of our investigation into the genetic basis of tumor cell radioresponse, we have isolated several clones with a wide range of responses to X-radiation (XR) from an unirradiated human colorectal tumor cell line, HCT116. Using human cDNA microarrays, we recently identified a novel gene that was down-regulated by two-fold in an XR-resistant cell clone, HCT116 Clone2_XRR . We have named this gene as X-ray radiation resistance associated 1 (XRRA1) (GenBank BK000541). Here, we present the first report on the molecular cloning, genomic characterization and over-expression of the XRRA1 gene.

Abstract. We previously isolated several clones that were closely-related genetically from a human colorectal tumor (HCT116) cell line. These clones displayed significantly different X-radiation response phenotypes. In this paper, we investigated how a single dose of X-radiation modulated the transcriptomic profiles of either the radiation-resistant (HCT116 Clone2_XRR ) or the radiation-sensitive (HCT116 CloneK_XRS ) clone when each was compared to a reference clone, HCT116Clone10_control . The latter represented a control clone that displayed a similar X-radiation response as the parental HCT116 cells. Pooled RNAs were obtained from HCT116 Clone2_XRR , HCT116 CloneK_XRS or HCT116 Clone10_control cells either before or at 10 min, 6 or 24 h after treatment with 4-Gy X-radiation. Transcriptomic profiles were assessed by cDNA microarrays. At least three independent experiments were carried out for each time point and statistical analysis was performed by paired t-test (p<0.05). From 19,200 genes/ ESTs examined, we identified only 120 genes/ESTs that were differentially expressed at any one of these four time points. Interestingly, different patterns of gene modulation were observed between the radiation-sensitive and radiationresistant clones. However, the fold changes of gene modulation were generally small (2-3 fold). Surprisingly, only 12.7% of 79 genes involved in DNA damage sensor/repair and cell cycle and between 2.6 and 9.2% of 76 genes involved in apoptosis, were significantly modulated in these early time points following irradiation. By comparison, up to 10% of 40 known housekeeping genes were differentially expressed. Thus in our experimental model, we were able to detect the up-regulation or down-regulation of mostly novel genes and/or pathways in the acute period (up to 24 h) following a single dose of 4-Gy X-radiation.