Isolation and Characterization of Stem Cells from Human Central Nervous System Malignancies

Khan, Imad Saeed; Ehtesham, Moneeb

doi:10.1007/978-3-319-16537-0_3

Cited by 3 publications

(4 citation statements)

References 93 publications

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“…Immunohistochemistry. The tumor tissues were fixed in 4% paraformaldehyde, washed with PBS, transferred to 70% ethanol and then embedded in paraffin in accordance with standard procedures (22). The antibodies used in this study were a goat anti-human Cx43 monoclonal antibody (ab87645, 1:2,000; Abcam Trading Company Ltd.) and mouse anti-human SUMO1 monoclonal antibody (sc-5308, 1:400; Santa Cruz Biotechnology).…”

Section: Methodsmentioning

confidence: 99%

“…To further investigate whether Cx43 SUMOylation increases GJIC in liver cancer stem cells, we observed the transmission of Hoechst 33342 in HepG2 cell spheres that were co-transfected with Cx43-wt alone or in combination with GFP-SUMO1. In theory, cancer stem cells are able to efflux the fluorescent dye, Hoechst 33342 (22). Therefore, fluorescence microscopic observations or flow cytometry should detect no staining.…”

Section: Cx43 Is Modified By Sumo1 In Liver Cancer Stem Cellsmentioning

confidence: 99%

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Connexin 43 SUMOylation improves gap junction functions between liver cancer stem cells and enhances their sensitivity to HSVtk/GCV

Shen

et al. 2018

Int J Oncol

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Connexin 43 (Cx43) can be modified and regulated by small ubiquitin-like modifier (SUMO)1; however, its role in liver cancer stem cells is poorly understood. In this study, we found a significant difference in the expression of Cx43 and SUMO1 between cancer stem cells and non-cancer stem cells in liver cancer. In liver cancer stem cells, Cx43 was almost absent, although the level of SUMO1 was significantly higher than that in non-cancer stem cells. Further experiments confirmed that the conjugated site of Cx43 by SUMO1 was located in Lys-144 and Lys-237, both of which are highly conserved among species. By the co-expression of Cx43 and SUMO1 in cancer stem cells, the gap junction intercellular communication (GJIC) of liver cancer stem cells was obviously improved. Using this feature, we verified whether it could effectively increase the sensitivity of cancer stem cells to the herpes simplex virus 1 thymidine kinase (HSVtk) gene in combination with ganciclovir (GCV), a conventional chemotherapeutic drug, in vitro and in vivo. As expected, increasing the expression of Cx43 SUMOylation in liver cancer stem cells effectively enhanced their sensitivity to HSVtk/GCV. On the whole, this study revealed a novel method which may be used to effectively restore GJIC in cancer stem cells in liver cancer, which enhances their sensitivity to conventional chemotherapeutic drugs.

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Section: Methodsmentioning

confidence: 99%

Section: Cx43 Is Modified By Sumo1 In Liver Cancer Stem Cellsmentioning

confidence: 99%

Connexin 43 SUMOylation improves gap junction functions between liver cancer stem cells and enhances their sensitivity to HSVtk/GCV

Shen

et al. 2018

Int J Oncol

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“…Glioblastoma Mutliforme (GBM) is the most common primary brain tumor and has a poor prognosis. The average survival rate is approximately 15 months even if the patient surgical resection combined with chemotherapy and radiotherapy [1]. Cell culture is a key technology for cancer research, as it allows scientists to study the biology of tumor cells in an environment with controlled variables [2].…”

Section: Introductionmentioning

confidence: 99%

Characterization of a new human astrocytoma cell line SHG140: cell proliferation, cell phenotype, karyotype, STR markers and tumorigenicity analysis

Sun

Chen

et al. 2021

J. Cancer

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Background: Primary tumor Cell was an important tool for tumor research. Here, a new astrocytoma cell line SHG-140 was established and its proliferation, phenotype, karyotype, STR authentication, pathological characteristics, and characteristics of the cells' intrancranial xenografts of nude mice were studied. Methods: Primary SHG-140 culture was performed in DMEM/F12 medium with 10% FBS. Cell proliferation, karyotype analysis, cell immunofluorescence and STR authentication of SHG140 cells were performed. HE staining and immunohistochemistry, Whole oncogene high flux sequencing of the patient sample were carried out. SHG140 cells were injected into the brain of nude mice, HE staining and immunohistochemistry of intracranial xenograft tumor were detected. Results: Cell immunofluorescence demonstrated that SHG140 cells were positive for A2B5 (Glial precursors ganglioside), GFAP (Glial fibrillary acidic protein), Nestin, S-100 (Acid calcium bingding protein), Olig2 (Oligodendrocyte transcription factor 2) and Ki67 (Nuclear-associated antigen), cells negatively stained for Vimentin. Cell proliferation curve revealed that SHG140 proliferated slightly within 48 h, which then significantly proliferated to the fourth day. Karyotype analysis demonstrated its total number of chromosomes was 55, having trisomy of chromosome 6, 7, 8, 9 and X, and tetrad of chromosome 1 and 21, chromosomal deletion and rearrangement were observed. STR markers analysis showed the cells were derived from human male. SHG140 cells had tumorigenic properties - the intracranial injection of these cells into nude mice gave rise to growing tumors. We found that the glioma tissue was diffusively positive for GFAP, Nestin, slightly positive for Olig2, S-100; the positive rate of Ki-67 was 65% and negative for Vimentin. SHG140 cells were tumorigenic, GFAP, Nestin, S-100 Olig-2, the proliferation marker Ki-67 were expressed in its intracranial xenograft, Vimentin was negative expressed. Whole oncogene high flux sequencing of the patient tissue showed TP53, PTEN, IDH1 and PTCH1 mutation were existed. Conclusions: Our study showed that SHG140 was an astrocytoma glioma continuous cell line derived from a human adult male, having a strong tumorigenicity in nude mice, which made it wound be a useful model for the study of human glioblastoma multiforme.

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“…A previous study revealed the existence of tumor stem cells (TSCs), which have the same functions as neural stem cells inside tumor cells including the ability to self-proliferate (3). There are a number of similarities between TSCs and neural stem cells that suggest the origin of TSCs (4), a variety of solid tumors, including brain tumors (5) and expression of numerous similar proteins (6). The discovery of TSCs has provided novel options for the treatment of brain tumors (7), including therapies targeting specific signaling pathways and transcription factors, and niche therapies targeting cancer stem cells (8,9).…”

Section: Introductionmentioning

confidence: 99%

Effects of NOTCH1 signaling inhibitor γ‑secretase inhibitor II on growth of cancer stem cells

Ding

Wang

et al. 2018

Oncol Lett

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The present study aimed to observe the effect of the Notch1 signaling inhibitor γ-secretase inhibitor II (GSI II) on the growth and differentiation of tumor cells. The tumor cell line U87 was grown in serum-free media, and cell growth was evaluated using immunofluorescence. Single-cell wall-adherent growing conditions were prepared, GSI II was added, and the differentiation and growth of single tumor cells was evaluated. Immunofluorescence demonstrated positive results for the expression of Nestin and cluster of differentiation 133. The cell proliferation rate was reduced following the addition of GSI II (P<0.05). GSI II may significantly inhibit the proliferation and differentiation of U87 tumor stem cells.

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Isolation and Characterization of Stem Cells from Human Central Nervous System Malignancies

Cited by 3 publications

References 93 publications

Connexin 43 SUMOylation improves gap junction functions between liver cancer stem cells and enhances their sensitivity to HSVtk/GCV

Connexin 43 SUMOylation improves gap junction functions between liver cancer stem cells and enhances their sensitivity to HSVtk/GCV

Characterization of a new human astrocytoma cell line SHG140: cell proliferation, cell phenotype, karyotype, STR markers and tumorigenicity analysis

Effects of NOTCH1 signaling inhibitor γ‑secretase inhibitor II on growth of cancer stem cells

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