Davy T. Lee

Identification of a novel function of Twist, a bHLH protein, in the development of acquired taxol resistance in human cancer cells

Taxol is one of the widely used chemotherapeutic drugs against many types of human cancer. While it is considered as one of the most effective anticancer drugs, treatment failure often occurs due to development of acquired resistance. Therefore, it is important to understand the molecular mechanisms responsible for the development of drug resistance. Although it is generally believed that taxol induces cell death through interfering with microtubules leading to mitotic arrest, recent evidence has suggested that taxol-induced cell death also occurs through pathways independent of either microtubule or mitotic arrest. In this study, we report the identification of a novel role for TWIST, a basic helix–loop–helix protein, which plays a central role in cell type determination and differentiation, during generation of acquired resistance to taxol in a nasopharyngeal carcinoma cell line, HNE1-T3, using comparative genome hybridization (CGH) and subsequent RT–PCR and Western blotting. We found that upregulation of TWIST was associated with cellular resistance to taxol but not other drugs with different mechanisms of action. The fact that increased TWIST protein levels were also associated with another microtubule-targeting anticancer drug, vincristine, in four types of human cancer including nasopharyngeal, bladder, ovarian and prostate, indicates that it may play a central role in the resistance to microtubule-disrupting agents. In addition, ectopic expression of TWIST into human cancer cells also led to increased resistance to both taxol and vincristine. Our results indicate a novel mechanism that leads to resistance to microtubule-disrupting anticancer drugs through upregulation of TWIST. Our evidence provides a therapeutic strategy to overcome acquired resistance through inactivation of TWIST expression in human cancer.

Decreased adhesiveness, resistance to anoikis and suppression of GRP94 are integral to the survival of circulating tumor cells in prostate cancer

The presence of circulating tumor cells (CTC) is common in prostate cancer patients, however until recently their clinical significance was unknown. The CTC stage is essential for the formation of distant metastases, and their continuing presence after radical prostatectomy has been shown to predict recurrent or latent disease. Despite their mechanistic and prognostic importance, due both to their scarcity and difficulties in their isolation, little is known about the characteristics that enable their production and survival. The aim of this study was to investigate the molecular mechanisms underlying the survival of CTC cells. A novel CTC cell line from the bloodstream of an orthotopic mouse model of castration-resistant prostate cancer was established and compared with the primary tumor using attachment assays, detachment culture, Western blot, flow cytometry and 2D gel electrophoresis. Decreased adhesiveness and expression of adhesion molecules E-cadherin, β4-integrin and γ-catenin, together with resistance to detachment and drug-induced apoptosis and upregulation of Bcl-2 were integral to the development of CTC and their survival. Using proteomic studies, we observed that the GRP94 glycoprotein was suppressed in CTC. GRP94 was also shown to be suppressed in a tissue microarray study of 79 prostate cancer patients, indicating its possible role in prostate cancer progression. Overall, this study suggests molecular alterations accounting for the release and survival of CTC, which may be used as drug targets for either anti-metastatic therapy or the suppression of latent disease. We also indicate the novel involvement of GRP94 suppression in prostate cancer metastasis.

Evidence of a novel docetaxel sensitizer, garlic-derived S-allylmercaptocysteine, as a treatment option for hormone refractory prostate cancer

The recent introduction of docetaxel in the treatment of hormone refractory prostate cancer (HRPC) has made a small but significant impact on patient survival. However, its effect is limited by intolerance and resistance. The aim of our study was to investigate if the garlic‐derived compound, S‐allylmercaptocysteine (SAMC), was able to act as a docetaxel sensitizing agent. First, the effect of SAMC on docetaxel sensitivity was examined on 3 HRPC cell lines by colony forming assay. We found that SAMC increased the efficacy of docetaxel on colony forming inhibition by 9–50% compared to single agent treatment. Second, using the HRPC CWR22R nude mice model, we found that the combination of SAMC and docetaxel was 53% more potent than docetaxel alone (p = 0.037). In addition, there was no additive toxicity in the mice treated with the combination therapy evidenced by histological and functional analysis of liver, kidney and bone marrow. These results suggest that SAMC is able to increase the anticancer effect of docetaxel without causing additional toxic effect in vivo. Third, flow cytometry and Western blotting analysis on HRPC cell lines demonstrated that SAMC promoted docetaxel‐induced G2/M phase cell cycle arrest and apoptotic induction. In addition, immunohistochemistry on CWR22R xenograft revealed a suppression of Bcl‐2 expression and upregulation of E‐cadherin in the SAMC and docetaxel treated animals. These results suggest that SAMC may promote docetaxel‐induced cell death through promoting G2/M cell cycle arrest and apoptosis. Our study implies a potential role for SAMC in improving docetaxel based chemotherapy for the treatment of HRPC.

Changes in Serum and Tissue Zinc Levels in Sex Hormone-Induced Prostatic Carcinogenesis in the Noble Rat

We investigated the changes in serum and tissue zinc levels in the Noble rat prostate gland under different pathological conditions induced by the administration of a combination of testosterone and 17β-estradiol. The results showed that there were significant differences in serum zinc values between normal and hormone-treated rats with prostatic hyperplasia, dysplasia and prostatic carcinoma (p < 0.05), although there was no significant difference among rats with different forms of prostatic lesions (i.e. hyperplasia, dysplasia and prostatic adenocarcinoma). There was also a difference in zinc content between the lateral prostate (LP), ventral prostate (VP) and dorsal prostate (DP) in normal rats. The zinc levels of LP were several times greater than those of either VP or DP (p < 0.01). There was also a great difference in zinc levels between the normal and the hyperplastic/dysplastic and carcinomatous LP and VP (p < 0.05). The levels of zinc in both LP and VP were increased in hyperplasia/dysplasia and carcinoma. On the other hand, the zinc levels of LP were much higher than those of VP in hyperplasia/dysplasia and carcinoma (p < 0.01), which may be correlated with the incidence of prostate cancers in these lobes (i.e. higher in LP and much lower in VP). In contrast, in DP, the levels of zinc were not affected, which may be correlated with the very low incidence of carcinoma in this lobe. Our data suggest that the difference in zinc levels among these lobes reflect the heterogeneity in zinc content in various lobes of the rat prostate. The growth and development of prostatic lesions in LP and VP may be positively correlated with the significant increase in tissue zinc levels in these lobes. On the other hand, the lack of response of DP to carcinogenesis may be due to its relatively stable low zinc levels. It is suggested that tissue zinc content may be used as a marker for prostatic lesions, including hyperplasia, dysplasia and carcinoma, while serum zinc levels may be a useful indicator for abnormal prostatic growth.