Characterization of a side population of astrocytoma cells in response to temozolomide

Chua, Constance; Zaiden, Norazean; Chong, K T; See, Siew Ju; Wong, Michael; Ang, Beng-Ti; Tang, Carol

doi:10.3171/jns/2008/109/11/0856

Cited by 66 publications

(60 citation statements)

References 58 publications

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“…With regards to the sorting techniques, there are two basic methods available, one is immunomagnetic beads or fluorescent activated cell sorting (FACS) based on the surface marker CD133; and the other method is the SP cell sorting technique based on a drug-resistant gene, such as an ATP-binding cassette or a multidrug resistance gene (MDR) (17,20). In either case, the supposed BGSCs are sorted, then identified and cultured.…”

Section: Discussionmentioning

confidence: 99%

A simplified and modified procedure to culture brain glioma stem cells from clinical specimens

et al. 2011

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Abstract. In recent years, the theory of brain glioma stem cells (BGSCs) has facilitated the study of gliomas. BGSCs have been accepted as the origin of gliomas and determine their biological features. Numerous efforts have been made to probe into the biological characteristics and behaviors of BGSCs. However, the culturing of target cells remains the essential first step for research on BGSCs. In this study, we established a simplified procedure to culture and isolate BGSCs from samples of clinical glioma patients. Samples of 17 glioma patients were included in the study, and the processed glioma cells were grown in serum-free stem cell media. After the tumor spheres appeared, a proliferation assay, a single-cellderived colonies formation assay and an induced differentiation assay were carried out, followed by an immunocytochemistry assay. Serial passage was used to purify the target cells, whereas neither animal experiments nor sorting techniques were included. As a result, CD133 + BGSCs from 8 out of 17 patients were grown and maintained in a serum-free condition combined with EGF, FGF and B-27 supplements. The tumor sphere cells were serially passaged and showed pluripotency in an induced differentiation assay. Immunocytochemistry identified the committed markers (CD133, and confirmed the cells were BGSCs and their progeny. The results proved that CD133 + BGSCs from resected glioma tissue may be cultured in serum-free stem cell media, and may also be purified by conditioned culture combining serial passage, which is time-saving and cost-effective, and allows the cells to be used for subsequent research. The cell sorting techniques and animal experiments of tumorigenecity are optional. Thus, this modified procedure is more practical and feasible than other available procedures.

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Section: Discussionmentioning

confidence: 99%

A simplified and modified procedure to culture brain glioma stem cells from clinical specimens

et al. 2011

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“…Several mechanisms are thought to account for such resistance, and DNA repair-related genes such as MGMT, MSH2, and MSH6 have been identified as key players involved in tumor survival after treatment with alkylating agents (1,(3)(4)(5). Moreover, a small population of cells with self-renewal capacity and immature phenotype, called "glioma stem cells," have been shown to be highly resistant to chemotherapy and radiation (6)(7)(8). In fact, several authors have hypothesized that these tumor stem cells are the source of the recurrent tumors after treatment.…”

Section: Introductionmentioning

confidence: 99%

“…In fact, several authors have hypothesized that these tumor stem cells are the source of the recurrent tumors after treatment. The phenotype, ex vivo assays and markers that define such stem cells are debated topics (6)(7)(8)(9). Bao et al used CD133 as a putative marker for characterization of these treatmentresistant cells in the setting of radiotherapy (8).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of Sonic Hedgehog and Notch Pathways Enhances Sensitivity of CD133+ Glioma Stem Cells to Temozolomide Therapy

Ulasov

Nandi

Dey

et al. 2010

Mol Med

168

118

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“…Some recent studies revealed that cellular stresses like drug exposure (Chua et al, 2008;Calcagno et al, 2010;Fong et al, 2010), hypoxia (Bar et al, 2010) and UV lighting (Liang et al, 2010) enriched for a CSC phenotype in cancer cell lines. In this study, we detected 1% cells positive for SP in the primary line SW1116 and observed a remarkable increase to 6%, 44% and 97% after HCPT treatment to 3 months, 6 months and 41 years, respectively.…”

Section: Discussionmentioning

confidence: 99%

205 MicroRNA Expression Profiling Identifies MiR-328 Regulates Cancer Stem Cell-Like SP Cells in Colorectal Cancer

Xu¹,

Xu²,

Tong³

et al. 2012

Gastroenterology

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BACKGROUND: Side population (SP) cells and their relationship to stem cell-like properties have been insufficiently studied in colorectal cancer (CRC). MicroRNAs (miRNAs) have attracted much attention but their roles in the maintenance of SP phenotype remain unclear. METHODS: The SPs from CRC cell lines and primary cell cultures were analysed for stem cell-like properties. MiRNA microarray analysis identified miR-328 as a potential stemness miRNA of SP phenotype. The level of miR-328 expression in clinical samples and its correlation with SP fraction were determined. Gain-of-function and loss-of-function studies were performed to examine its roles in cancer stem-like SP cells. Furthermore, bioinformatics prediction and experimental validation were used to identify miR-328 target genes. RESULTS: The SP cells sorted from CRC possess cancer stem cell (CSC)-like properties, including self-renewal, differentiation, resistance to chemotherapy, invasive and strong tumour formation ability. MiR-328 expression was significantly reduced in SP cells compared with Non-SP cells (Po0.05). Moreover, miR-328 expression was downregulated in CRC (n ¼ 33, Po0.05) and low miR-328 expression tend to correlate with high SP fraction (n ¼ 15, r ¼ 0.6559, Po0.05, Pearson's correlation). Functional studies indicated that miR-328 expression affects the number of SP cells. In addition, miR-328 overexpression reversed drug resistance and inhibited cell invasion of SP cells. Furthermore, luciferase reporter assay demonstrated that miR-328 directly targets ABCG2 and MMP16 and affects the levels of mRNA and protein expression in SP cells. CONCLUSION: These findings indicate that CRC contain cancer stem-like SP cells. MiR-328 has an important role in maintaining cancer stem-like SP phenotype that may be a potential target for effective CRC therapy.

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Characterization of a side population of astrocytoma cells in response to temozolomide

Abstract: Knockdown of ABCG2 transporters did not abrogate the SP cell response to temozolomide. Upregulation of several other ABC drug transporter genes is proposed to account for this chemoresistance.

Cited by 66 publications

References 58 publications

A simplified and modified procedure to culture brain glioma stem cells from clinical specimens

A simplified and modified procedure to culture brain glioma stem cells from clinical specimens

Inhibition of Sonic Hedgehog and Notch Pathways Enhances Sensitivity of CD133+ Glioma Stem Cells to Temozolomide Therapy

205 MicroRNA Expression Profiling Identifies MiR-328 Regulates Cancer Stem Cell-Like SP Cells in Colorectal Cancer

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