Effect of ectopic expression of decorin in a leukemic cell line engineered to express TAL1 and LMO1 proteins

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Kamara, Kandeh.
Olesen, James B.
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Thesis (M.S.)
Department of Biology
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Progress in understanding cancer progression has been hampered over the years by the different types of mutations present and irregular changes of gene regulation associated with any given cancer. In this work, molecular interactions between TALI, LMO1, and decorin were investigated. Numerous studies have shown that ectopic expression of decorin protein induced growth suppression in neoplastic cells of various histogenic origins. Furthermore, ectopic expression of TAL1 and LMOI oncoproteins has been shown to occur in approximately 50% of the cases of T-cell acute lymphoblastic leukemia (T-ALL). It was of interest then to determine the preventive or interactive role decorin played with the oncogenic activity of TAL I and LMO1. In this investigation, decorin was introduced into a murine T-cell line (AKR-DP-603) through the use of the mammalian expression vector pcDNA3.1 (-). This particular cell line was previously engineered to express TALI and LMO1. Protein expression patterns in all cell populations were analyzed using the Western blot technique and a proteoglycan with a molecular weight of 100 kDa before chondroitinase ABC treatment and a core protein of55 kDa after treatment with chondroitinase ABC was seen. This finding is significant since it implies that the pcDNA3. 1(-) vector containing decorin cDNA was present, and the corresponding decorin peptides were expressed in both cytoplasmic and nuclear extracts. Furthermore, Northern blot analysis was performed on total RNA extracts to determine the transcriptional state of endogenous decorin rRNA, as well as exogenously introduced decorin. Northern blot analysis revealed no decorin-specific mRNA transcripts from the various cell populations. This result did not imply a lack of possible regulatory effect on protein and mRNA levels of TALL and LMOI by decorin. Finally, cell growth assays were performed on all cell populations and cell counts were used to assess the growth pattern of each population after serum withdrawal. The results show possible growth suppressive effects of decorin on TAL1 and LMOI expressing cells. Results obtained from this study shed further light on the molecular interactions influencing T-ALL and may also help in the design of potentially beneficial cancer treatments using decorin.