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Rapid Detection of Lectin Receptors on Human Cancer Cells
Author(s) -
Welty Lily Anne Y.,
Heinrich Eileen L.,
Garcia Karina,
Banner Lisa R.,
Summers Michael L.,
Baresi Larry,
Metzenberg Stan,
CoyleThompson Cathy,
Oppenheimer Steven B.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1105-c
Subject(s) - lectin , receptor , cancer cell , microbiology and biotechnology , cell , chemistry , ligand binding assay , cell culture , fluorescence , biochemistry , biology , cancer , genetics , physics , quantum mechanics
This laboratory has developed novel assays and other procedures for rapidly detecting surface receptors and ligands. Here, we use two human colon cell lines, CCL‐220 cancer, and CRL‐1459 non‐cancer, obtained from ATCC. We compared the binding of lectins from 3 species (Triticum vulgaris, Phaseolus vulgaris and Lens culinaris) to these cell lines using two different assays: a lectin derivatized bead binding assay and a FITC‐ lectin binding assay. We used 3 different cell preparation procedures for these cell types: (1) frozen, thawed and formaldehyde fixed; (2) freshly cultured and formaldehyde fixed; and (3) live. The fluorescence and bead binding results, including haptenic inhibition studies with preferential and non‐preferential binding sugars (0.2 M), were nearly identical. The cancer cells bound well to the 3 lectins, while the non‐cancer cells did not. The fluorescence assay was able to detect lower lectin receptor levels than the bead assay, while the bead assay was much easier to do and was far more rapid than the fluorescence assay. All 3 cell preparation procedures yielded cells that bound the lectins in the same manner. Most important is the finding that bead binding experiments with the frozen, thawed and fixed cells was found to be the method of choice for the most rapid detection of cell surface receptors without the requirement for costly and time consuming cell culture (supported by NIH NIGMS SCORE, RISE, MARC, ITQ program and the Joseph Drown Foundation).