Abstract
Frizzled-7 protein plays a significant role in the formation of several malignant tumors. Up regulation of the Frizzled-7 in cancer cell lines is associated with nuclear accumulation of wild-type β-catenin from the Wnt/β-catenin pathway which is frequently activated in tumors. To analyze activity of the Frizzled-7 promoter in tumor cells, we constructed two recombinant plasmid vectors in which the Frizzled-7 promoter was used to drive the expression of green fluorescent protein (GFP) and Shiga-like toxin I (Stx1) (pFZD7-GFP/Stx1) genes. The Frizzled-7 protein was found to be expressed in the cancer cell lines but not in the normal cell lines. The GFP expression was restricted to the cancer cell lines and xenografts in the BALB/C mice but not to normal cell lines. Moreover, cell proliferation and tumor growth decreased significantly after transfection with the pFZD7-Stx1. Results from this study will help determine a highly effective strategy for gene therapy of tumors.
Highlights
Cancer represents a leading health problem worldwide and surgery is not a treatment option in many cases, because of late diagnosis and poor therapeutic options available
Using reverse-transcriptase polymerase chain reaction (RT-PCR), researchers found that the Wnt mRNAs were over expressed in the hepatocellular carcinoma (HCC) cells, where they detected the over expression of Wnt3, Wnt5A, Wnt6, and Wnt11 among the 19 studied human Wnt ligands [5]
The luciferase gene of pFZD7Luc was replaced with the green fluorescent protein (GFP) or Stx1 genes using the NcoI and XbaI restriction endonucleases (Figure 1B) and the fidelity of the plasmids was confirmed by sequencing analysis
Summary
Cancer represents a leading health problem worldwide and surgery is not a treatment option in many cases, because of late diagnosis and poor therapeutic options available. Gene therapy represents an attractive approach for treatment of cancers and other chronic diseases. Members of the Frizzled family of seven-pass transmembrane proteins serve as receptors for the Wnt signaling glycoproteins. The Wnts are a family of secreted glycoproteins that serve as extracellular signaling molecules involved in cell differentiation, migration, and proliferation during embryonic development. These proteins cause tumor formation when they are aberrantly activated [4,5,6]. Using reverse-transcriptase polymerase chain reaction (RT-PCR), researchers found that the Wnt mRNAs were over expressed in the HCC cells, where they detected the over expression of Wnt, Wnt5A, Wnt, and Wnt among the 19 studied human Wnt ligands [5]
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