Abstract 1609: Functional analysis of SFRS10: A novel molecular target for anti-cancer drug

Abstract

Abstract [Background] Development of novel molecular-target drugs is actively conducted: A large number of such drugs are under clinical trials. However, an ideal molecular-target drug with high-specificity and low-toxicity is yet to be developed. For epoch-making development of cancer drugs, it is necessary to further identify target candidates by unprecedented approach. [Purpose] Last year, we reported 13 candidate genes that contributed to an immortalization through comprehensive gene expression analysis of human mortal- and immortal-cells and tissues. Under the same procedure, we here added SFRS10 gene to the list. To evaluate its possibility as drug target, we attempted here a functional analysis of SFRS10. [Materials and Methods] Human colon cancer cell lines DLD-1 and HCC48 and pancreatic cancer cells MIAPaCa-2, KLM-1, and BxPC-3 were used in each experiment. Quantitative analyses of gene expression levels were conducted by real-time RT-PCR method using gene specific-primers and Universal Probe Library probes (Roche). Microarray analysis was conducted with 4×44K Whole Human Genome Microarray (Agilent). Pathway analysis was performed through the use of Ingenuity Pathways Analysis (IPA, Ingenuity® Systems, www.ingenuity.com). Knockdown experiments were performed using Stealth RNAi and RNAiMax reagent (Invitrogen). Cell proliferation was measured with MTT assay. Caspase activities were analyzed using ApoAlert Caspase Profiling Plate (Clontech). Cell cycle profile was obtained by measuring PI-stained cells with FACS “Calibur” (BD Biosciences). [Results] SFRS10 knockdown induced inhibition of cell proliferation of all the tested cells. Because knockdown efficiencies and cytostatic activities showed a good correlation in the experiment, we speculated that SFRS10 may have an important role for the cell proliferation. Treatment with SFRS10 siRNA induced cell cycle arrest in the S and G2/M phases and apoptosis in DLD-1 and KLM-1 cells. Moreover, we observed that some cell cycle-related genes, such as MAD2L1 and RFC4, altered their expression levels in response to SFRS10 knockdown as assessed by microarray analysis; cell cycle checkpoint pathways were significantly changed with network analyses. The alteration of gene expression levels were confirmed by real-time RT-PCR analyses. [Conclusion] These results suggested that SFRS10 may have an important role in regulation of cell proliferation and may control cell cycle checkpoints. Taken together, SFRS10 may be one molecule responsible for immortalization of cancer cells and the useful molecular target for cancer therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1609.