Cancer stem cells: Cell culture, markers, and targets for new therapies

Gilbert, Candace A.; Ross, Alonzo H.

doi:10.1002/jcb.22350

Cited by 165 publications

(178 citation statements)

References 70 publications

(83 reference statements)

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“…CSCs play an important role in tumor progression (3,15). To confirm whether the effect of forced Nanog expression on HeLa cell malignant behavior is associated with CSCs, we tested the tumor sphere forming ability of HeLa-Nanog cells.…”

Section: Discussionmentioning

confidence: 99%

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Forced expression of Nanog with mRNA synthesized in vitro to evaluate the malignancy of HeLa cells through acquiring cancer stem cell phenotypes

Ding

Wang

et al. 2016

Oncology Reports

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Abstract. Nanog is a pluripotency-related factor. It was also found to play an important role in tumorigenesis. To date, the mechanisms underlying cervical tumorigenesis still need to be elucidated. In the present study, Nanog mRNA was synthesized in vitro and transfected into HeLa cells. After mRNA transfection, the forced expressed of Nanog in HeLa cells led to markedly increased invasion, migration, resistance to chemotherapeutic agents and dedifferentiation. In a subcutaneous xenograft assay, these cells had significantly increased tumorigenic capacity. Real-time PCR indicated that Nanog-induced dedifferentiation was associated with increased expression of endogenous Oct4, Sox2 and FoxD3. In addition, the dedifferentiated HeLa cells acquired features associated with cancer stem cells (CSCs), such as multipotent differentiation capacity, and expression of CSC markers such as CD133. These data imply that Nanog is a positive regulator of cervical cancer dedifferentiation.

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Section: Discussionmentioning

confidence: 99%

“…Cancer stem cells (CSCs) appear to be responsible for tumor initiation, progression and resistance to conventional treatment (2,3). Nanog is one of the core transcription factors for maintaining the stemness of embryonic stem cells (ESCs) (4).…”

Section: Introductionmentioning

confidence: 99%

Forced expression of Nanog with mRNA synthesized in vitro to evaluate the malignancy of HeLa cells through acquiring cancer stem cell phenotypes

Ding

Wang

et al. 2016

Oncology Reports

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show abstract

“…However, it has been demonstrated that they be isolated by several ways including surface marker sorting (Al-Hajj et al, 2003;Singh et al, 2004), side-population cells selecting (Hadnagy et al, 2006;Moserle et al, 2010), and mammosphere cultures (Ponti et al, 2005;Yu et al, 2008). Most CSCs form clones when nurtured in low adhesive vessels (Chumsri and Burger, 2008;Kobayashi et al, 2008;Gilbert and Ross, 2009), and further generate sphere clones in the present of growth factors such as epidermal growth factor (EGF). Therefore the sphere clone formation assay has been widely used for the separation and identification of CSCs (Ponti et al, 2005;Yu et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD

Yin

Zong

et al. 2011

J. Zhejiang Univ. Sci. B

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Abstract:This paper aims to screen and identify sphere clone cells with characteristics similar to cancer stem cells in human gallbladder cancer cell line GBC-SD. GBC-SD cells were cultured in a serum-free culture medium with different concentrations of the chemotherapeutic drug cisplatin for generating sphere clones. The mRNA expressions of stem cell-related genes CD133, OCT-4, Nanog, and drug resistance genes ABCG2 and MDR-1 in sphere clones were detected by quantitative real-time polymerase chain reaction (PCR). Stem cell markers were also analyzed by flow cytometry and immunofluorescent staining. Different amounts of sphere clones were injected into nude mice to test their abilities to form tumors. Sphere clones were formed in serum-free culture medium containing cisplatin (30 μmol/L). Flow cytometry results demonstrated that the sphere clones expressed high levels of stem cell markers CD133 + (97.6%) and CD44 + (77.9%) and low levels of CD24 + (2.3%). These clones also overexpressed the drug resistance genes ABCG2 and MDR-1. Quantitative real-time PCR showed that sphere clones expressed stem cell genes Nanog and OCT-4 284 and 266 times, respectively, more than those in the original GBC-SD cells. Immunofluorescent staining showed that sphere clones overexpressed OCT-4, Nanog, and SOX-2, and low expressed MUC1 and vimentin. Tumor formation experiments showed that 1×10 3 sphere clone cells could induce much larger tumors in nude mice than 1×10 5 GBC-SD cells. In conclusion, sphere clones of gallbladder cancer with stem cell-like characteristics can be obtained using suspension cultures of GBC-SD cells in serum-free culture medium containing cisplatin.

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“…Due to possessing a strong drug tolerance, CSCs are not easily destroyed, which leads to the failure of treatment. Therefore, a key goal for tumor therapy is to effectively induce the apoptosis of tumor stem cells (23).…”

Section: Discussionmentioning

confidence: 99%

Antitumor effects of matrine on cancer stem like cells isolated from the human liver cancer SMMC‑7721 cell line

et al. 2017

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Abstract. The existence of cancer stem cells (CSCs) or cancer stem-like cells (CSLCs) is regarded as the cause of tumor formation and recurrence. Matrine has been reported to exhibit antitumor effects in cancer cells. In the present study, a preliminary study was performed on the mechanisms of matrine on hepatocellular carcinoma (HCC) stem-like cells. The HCC SMMC-7721 cell line was cultured in tumor stem cell-specific medium to form spheres, and different concentrations (1, 2 and 5 mg/kg) of cisplatin were then used in order to purify the most drug-resistant cells, which were used as CSLCs. An MTT assay was performed to detect the inhibitory effects of matrine against CSLC proliferation. Quantitative polymerase chain reaction (qPCR) and western blot analysis were used to detect changes in cell adhesion regulating gene (CAR), E-cadherin, laminin and fibronectin. As a result, using tryptose sulfite cycloserine medium culture and cisplatin-resistance screening, CSLCs were successfully isolated from the SMMC-7721 cell line. Matrine inhibited the proliferation of CSLCs in vitro. The results of qPCR and western blot analysis demonstrated that matrine upregulated the expression of CAR, E-cadherin, laminin and fibronectin in CSLCs compared with the control treatment. A certain concentration of matrine exhibited antitumor effects on HCC stem like cells.

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Cancer stem cells: Cell culture, markers, and targets for new therapies

Cited by 165 publications

References 70 publications

Forced expression of Nanog with mRNA synthesized in vitro to evaluate the malignancy of HeLa cells through acquiring cancer stem cell phenotypes

Forced expression of Nanog with mRNA synthesized in vitro to evaluate the malignancy of HeLa cells through acquiring cancer stem cell phenotypes

Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD

Antitumor effects of matrine on cancer stem like cells isolated from the human liver cancer SMMC‑7721 cell line

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