Cancer Res

Cancer Res. cells. These findings suggested that SAHA treatment created a more drug-resistant state in residual ALDHbr cells. The imaging technique may facilitate searching and characterization of CSCs. by non-adherent suspension culture in serum-free medium, and they have been widely used to study underlying key molecular pathways [3]. Mounting evidences suggest that the tumor microenvironment is responsible for conditioning the stem cell status itself. The system has been questioned because of the entire differences between and systems in microenvironment. Side population (SP) technique and flow cytometry using cell surface markers have been applied to isolate CSC, but the specificity of these two methods is usually under debate. Previous studies reported that non-SP cells and CD133 cells can also generate tumors in NOD/SCID mice [4, 5]. Regarding the limitations in the isolation procedures, especially those used stem cell surface markers, would result in CSC injury, we designed an method using intracellular markers of stem cells which were identified in various human cancers to isolate CSCs from xenograft tumors in animal model. Aldehyde dehydrogenases (ALDHs) are detoxifying enzymes within a superfamily. In fact, Adamts5 the expression level of ALDH in stem cells usually high enough to protect them against oxidative insult, suggesting their well-known longevity. ALDH converts retinol to retinoic acid, a modulator of cell and stem cell proliferation. Elevation of the level of ALDH activity has been seen in stem cell populations of breast cancer [6], lung cancer [7], liver cancer [8] and colon cancer [9]. An ALDEFLUOR kit (Stem Cell Technologies) designed for precise identification and isolation of ALDH-bright CSCs using specific inhibitor for ALDH activity diethylaminobenzaldefyde (DEAB) Sofalcone was thus applied in this study. Histone deacetylase inhibitors (HDACIs) can induce hyperacetylation of specific proteins, recently considered as a new solution to inhibit cell proliferation and promote differentiation of various hematologic and solid tumors [10, 11]. Suberoylanilide hydroxamic acid (SAHA, Vorinostat), an HDACI, was approved by FDA for treatment of cutaneous T-cell lymphoma in 2004 [12]. Recent investigations Sofalcone exhibited that SAHA treatment can suppress the expression of the stem cell marker CD133 in glioma [13]. In addition, SAHA can also inhibit the ability to proliferation, self-renewal, migration, and invasion in human pancreatic CSCs by up-regulation of miR-34a [14]. These results implied that SAHA could be a potential agent for the therapy against CSCs. However, some studies revealed that SAHA leads to the increase of the stem cell markers in epithelialCmesenchymal transitions (EMT) phenotypic prostate cancer cells [15, 16]. These findings are in consistent with the clinical results of HDACIs, which have shown promise efficacy in hematological malignancies while disappointed effects in epithelial cell-derived cancers. The detailed mechanism of this phenomenon remains to be elucidated. In the current study, we aim to determine the significance role of SAHA in the mediation of CSCs in lung cancer. The ALDEFLUOR assay and FACS analysis were used to isolate CSCs from human lung carcinoma grown as xenografts on nude mice. The results showed that SAHA retards the growth of H1299 xenografts and decreases CSC population, but induces EMT phenotype and activates pluripotency associated program in the residual CSCs. Our results provide a possible mechanism for the limited treatment response of HDACIs in the clinical trials around the epithelial cell-derived cancer. RESULTS SAHA enhances the expression of CSC characteristics was examined in the H1299 human non-small cell lung cancer xenograft, which was inoculated subcutaneously in nude mice. The Sofalcone tumor progression rate was assessed by luciferase bioluminescent imaging. In the treatment group, the signals in the tumor are significant lower than that in vehicle-treated control tumor (Fig. ?(Fig.2A).2A). Daily administration of SAHA with the dosage of 100 mg/kg/day caused significant suppression of the growth of established H1299 tumors; reduction of 63% tumor volume compared with that of the vehicle-treated control animals (Fig. ?(Fig.2B).2B). Each animal receiving 100 mg/kg/day SAHA survived for at least 10 days. These results indicate that SAHA effectively reduces the tumor Sofalcone growth of H1299 xenografts at the dose of 100 mg/kg/day. Open in a separate window Physique 2 SAHA effectively inhibits the growth of H1299 tumor cells(A) Bioluminescent images of mice bearing H1299-CMV-Luc tumors before and after SAHA treatment. (B) Tumor growth curve of the subcutaneous H1299-CMV-Luc lung cancer xenograft in mice Sofalcone daily treated with vehicle alone or SAHA (100 mg/kg, i.p.). Data are presented as the mean tumor volume S.E. of the surviving animals in each groups..