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BauernOpfer
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SleepyHollow02
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Untersuchte Arbeit:
Seite: 50, Zeilen: 1 ff.
Quelle: Gu et al 2009
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4. RESULTS

4.1 mAR expression in colon cancer cell lines

In my diplom thisis, while analyzing in vivo mAR expression in paraffin blocks generated from xenograft tumor tissues of various origins, we have noticed significant mAR expression in colon cancer xenograft specimens. In line with these findings mAR expression was subsequently detected by confocal laser scanning microscopy using the fluorescent testosterone-HSA-FITC conjugate in cultured HCT116- (Figure5 e,f), or in Caco2-colon cells (Figure 5a,b) while HSA-FITC labeled CaCo2 or HCT116 cells showed no apparent staining (Figures 5 c, d, g, h). These results indicate that mAR in expressed in colon cancer cell lines. Interestingly, mAR staining could not be detected in the non-transformed intestinal cell line IEC06 (Fig. 6A). These staining experiments and the fact that testosterone-HSA-FITC is an impermeable conjugate disclosed mAR expression preferentially in colon cancer cell lines and tumors. In addition, mAR could be also detected in iAR silenced of Caco2 cells by using testosterone-HSA-FITC. These results imply that the molecular identity of mAR is probably not identical with iAR (Fig. 6B)

Figure 5 Membrane staining of mAR in Caco2 and HCT 116 colon cancer cells

Results

mAR expression in specimens of colon tumors and colon cancer cell lines While analyzing paraffin blocks generated from in vivo xenograft tumor tissues of various origins, we noticed significant mAR expression in colon tumors. Specifically, using testosterone-HSA-FITC fluorescent conjugates we detected specific, FITC-related fluorescence in membrane specimens of colon xenograft tumors generated from wild type HCT116 cells (WCL2) (Fig. 1a,a') or HCT116 p53-/- cells (MCL3) (Fig. 1d,d'). Conversely, no apparent staining could be identified in control tissues labeled with HSA-FITC (Fig. 1c,f). Although the apparent visualization of mAR staining in tissue preparations is restricted by technical limitations, in cultured HCT116- (Fig. 1h) or in Caco2-colon cells (Fig. 2A a,b) the membrane staining of mARs was obvious by confocal laser scanning microscopy using the fluorescent testosterone-HSA-FITC conjugate. No apparent staining was evident in HSA-FITC-labeled HCT116 or Caco2 cells (Fig. 1j, 2Ac,d). Interestingly, mAR staining could not be detected in the membrane of the non-transformed intestinal cell line IEC06 (Fig. 2B). These staining experiments and the fact that testosterone- HSA-FITC is an impermeable conjugate disclosed mAR expression preferentially in colon cancer cell lines and tumors.

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