Downregulation of transcription factor SOX2 in cancer stem cells.pptVIP

Downregulation of transcription factor SOX2 in cancer stem cells suppresses growth and metastasis of lung cancer 2012-5-25 Publication: British Journal of Cancer Date: Jun 7, 2011 影响因子：4.831 Experimental Route Background The cancer stem cell hypothesis ： clones are maintained by a small subpopulation of cells——indefinite proliferation and differentiation potentials. Cancer stem cells ：efflux Hoechst 33342 dye —— ‘side population’ (SP). Background The links between SOX transcription factors and cancer. SOX2 —— an immunogenic antigen in 41% of small cell lung cancer patients. SOX2 —— invasion 、metastasis of prostate cancers. Molecular mechanisms ：SOX2 in breast cancer promotes cell proliferation and tumourigenesis by facilitating the G1/S transition. Background The network of SOX2: SOX2, Oct 4, Nanog, FGF-4, UTF1,Fbx15 and Lefty1. Altering the level of Oct 4 have provided useful insights into the developmental potential of ES and EC cells——how altering the levels of SOX2 influences cell fate. How to change SOX2 and SOX2/Oct 4 levels to influence CSCs. Can the CSC concept be applied to a mouse lung tumour model by identifying SP retaining low Hoechst 33342 dye in a murine D121 non-small cell lung carcinoma cell line? Does the SP display stem cell characteristics in vitro and highly express transcription factor SOX2? Dose the SiRNA-mediated knockdown of SOX2 in CSCs significantly suppress tumour growth and metastases? MATERIALS AND METHODS Animal cell lines and reagents The C57BL/6 mice, 6–8 weeks of age. The murine D121 non-small lung carcinoma cell line was provided by Dr Lea Eisenbach. The IFN-γand MHC-I antibody were purchased from Invitrogen. Flow cytometry analysis and cell sorting Reverse transcriptase (RT)–PCR analysis Total RNA from D121-SP and non-SP cell samples was isolated using RNAeasy. The RNA (D121) cells →cDNA RT–PCR ——SuperScript One-Step . Primers : ABCG2 、Sca-1 、Wnt-1 、Wnt-2 、Wnt-10a 、SOX2 、Oct 4 、Notch1 、GAPDH.