Contents

       Human lung adenocarcinoma cell A549 was transfected stably with the DNA construct, EGFP-hcrmp1, containing 1.9 kb of promoter region of CRMP-1 gene and the reporter (GFP) gene.  The promoter fragment was inserted into the multiple cloning site of pEGFP-1 plasmid (BD Biosciences; Ref. 1).

Product Information

      CRMP-1 promoter/GFP Reporter Cells are homogenous and show proportional increase of fluorescent intensity (Figure 2) during cell growth (Figure 1).  Microscopic analysis indicates that at least over 90% of GFP-expressing cells deliver detectable fluorescent signal (Figure 3).

Figure 1: Growth curve.	Figure 2: Fluorescence signal.

   Figure 3: Microscopic Images.

(Suspension)	(adherent Cells)

Applications

       Screening for agents or compounds, which could induce the expression of hCRMP-1.  The increase of GFP signal after treatment of CRMP-1 promoter/GFP Reporter Cells indirectly means the up-regulation of hCRMP-1 level.  This system will be very suitable for screening active agents, which might be involved in the regulation or related pathways.

Background information

       Human CRMP-1 was reported as “a lung cancer invasion suppressor gene with nerve” (Ref. 2).  Recent discovery showed that the level of expression of the gene encoding CRMP-1 inversely affects cancer invasion and metastasis, (i.e., the higher the level of expression, the lower the incidence of cancer invasion and metastasis) and thus characterized the CRMP-1 gene as an invasion-suppression gene (Ref. 3-5).  Further analysis of around 50 cases of clinical lung tissue samples using real time-quantitative PCR techniques found that low-expression patients of CRMP-1 had more advanced diseases and lymph node metastases, while high-expression patients of CRMP-1 had a significantly longer disease-free and overall survival period.  “CRMP-1 as an excellent target for lung cancer” was interpreted by the web “bioportfolio” (Ref. 6).

References

(1) pEGFP-1 (BD Biosciences; http://orders.clontech.com/clontech/techinfo/vectors_dis/pEGFP-1.shtml

(2) Steeg (2001) J. Natl. Cancer Inst. (2001), 93(18): 1364-1365

(3) Shih et al., J. Natl. Cancer Inst. (2001), 93(18): 1392-1400.

(4) Chu et al., Am. J. Respir. Cell Mol. Biol. (1997), 17:353-360.

(5) Shih et al., Clinical & Exper. Metastasis (2003) 20: 69-76).

(6) CRMP-1 as an excellent target for lung cancer

http://www.bioportfolio.com/LeadDiscovery/Pubmed-110112.htm