The crucial role of cancer stem cells (CSCs) in the pathology of malignant diseases has been extensively studied during the last decade Nestin, a class VI intermediate filament protein, was originally detected in neural stem cells during development Its expression has also been reported in different tissues under various pathological conditions Specifically, nestin has been shown to be expressed in transformed cells of various human malignancies, and a correlation between its expression and the clinical course of some diseases has been proved Furthermore, the coexpression of nestin with other stem cell markers was described as a CSC phenotype that was subsequently verified using tumorigenicity assays The primary aim of this review is to summarize the recent findings regarding nestin expression in CSCs, its possible role in CSC phenotypes, particularly with respect to capacity for self-renewal, and its utility as a putative marker of CSCs

https://onlinelibrary.wiley.com/doi/pdf/10.1111/cas.12691

Nestin as a marker of cancer stem cells

The challenge of targeting cancer stem cells to halt metastasis.

Side population (SP) cells are previously identified from bone marrow based on their capacity to efflux of the fluorescent dye Hoechst 33342. Recent studies demonstrate that SP cells isolated from various cancer cell lines and primary tumors possess stem-cell-like properties. Thus, targeting tumor SP cells may provide new strategies for treatment in clinic. We previously showed that 1,3,8-trihydroxy-6-methylanthraquinone (emodin), a reactive oxygen species (ROS) generator, enhanced sensitivity of gallbladder cancer SGC-996 cells to cisplatin (CDDP) via generation of ROS and downregulation of multidrug-resistance-associated protein 1 (MRP1). To determine whether emodin also acts effectively on cancer stem cells of gallbladder carcinoma, we use SP cells as a model of cancer stem-cell-like cells. Here, we found that emodin, via ROS-related mechanism and suppressing the function of ATP-binding cassette super-family G member (ABCG2), which is known to be associated with Hoechst dye efflux activity of SP cells, not only reduced the ratio, inhibited clone formation, and eliminated sphere formation of SP cells effectively, but also promoted obviously the intracellular accumulation of doxorubicin, the main substrate of the efflux pump ABCG2. In addition, emodin could sensitize CDDP, via inhibition of expression of ABCG2, to overcome chemoresistance of SP cells. Importantly, similar to the experiment in vitro, emodin/CDDP co-treatment in vivo suppressed the tumor growth derived from SP cells through downregulating ABCG2 expression. Our results suggest that emodin is an effective agent targeting cancer stem-like SP cells of gallbladder carcinoma, either alone or acts as a chemotherapy enhancer.

/pdf/emodin-as-an-effective-agent-in-targeting-cancer-stem-like-3l7corozo9.pdf

Emodin as an effective agent in targeting cancer stem-like side population cells of gallbladder carcinoma.

Tumor development has long been known to resemble abnormal embryogenesis. The embryonic stem cell gene NANOG , a divergent homeodomain transcription factor that is independent of leukemia inhibitory factor, has been reported to be expressed in germ cells and in several tumor types. However, the short-term expression and role of NANOG in cervical cancer remain unclear. In the present study, we demonstrate that NANOG exhibits cellular shuttling behavior and increasing stromal distribution during the progression of cervical cancer. Our molecular data using RT-PCR and restriction enzyme digestion show that NANOG is mainly transcribed from the NANOG gene in cervical cancer. In addition, IHC using confocal microscopy suggests that mesenchymal stem cells (MSCs) are one type of cytoplasmic NANOG-positive cells in cervical cancer stroma. Co-culture of cervical cancer–derived MSCs with SiHa cells showed increased proliferation characteristics in vitro and enhanced tumor growth in vivo . Our results show, for the first time to our knowledge, that MSCs are a source of cytoplasmic NANOG expression in the cervical cancer stroma and that they participate in the progression of cervical cancer both in vitro and in vivo . Our study provides evidence that NANOG is a cervical cancer progression marker and also serves as a starting point for a more extensive exploration of the cellular translocation of NANOG and the multifunctionality of the stromal microenvironment.

Cytoplasmic NANOG-Positive Stromal Cells Promote Human Cervical Cancer Progression

Context: Carcinoma of the gallbladder (GBC) is the most common biliary tree cancer in the world. Beside gallstones, no specific risk factors for GBC are currently established. Several published studies have identified various prognostic gene expression markers in GBC.Objective: The present article reviewed published studies on gene expression biomarkers and gallbladder cancer susceptibility.Methods: We searched the PubMed, Medline, and Embase databases using the search terms “Gallbladder”, “cancer/carcinoma”, “expression”, “genes”, “proteins”, and “biomarker” updated until June 2012 and limited to English language papers. The online searching was accompanied by checking reference lists from the identified articles for potentially eligible original reports.Results: Potential GBC biomarkers identified by different studies were summarized.Conclusion: To infer, the present article highlights a few potential biomarkers in GBC. However, none of the markers identified so far are effective as a routine screening ...

/pdf/potential-biomarkers-in-gallbladder-cancer-present-status-4u40gmzc3w.pdf

Potential biomarkers in gallbladder cancer: present status and future directions

AIM: To identify cancer stem cells (CSCs) in human gallbladder carcinomas (GBCs).

METHODS: Primary GBC cells were cultured under serum-free conditions to produce floating spheres. The stem-cell properties of the sphere-forming cells, including self-renewal, differentiation potential, chemoresistance and tumorigenicity, were determined in vitro or in vivo. Cell surface expression of CD133 was investigated in primary tumors and in spheroid cells using flow cytometry. The sphere-colony-formation ability and tumorigenicity of CD133+ cells were assayed.

RESULTS: In vitro culture experiments revealed that floating spheroids were generated from primary GBC cells, and these sphere-forming cells could generate new progeny spheroids in serum-free media. Spheroid cells were differentiated under serum-containing conditions with downregulation of the stem cell markers Oct-4, Nanog, and nestin (P < 0.05). The differentiated cells showed lower spheroid-colony-formation ability than the original spheroid cells (P < 0.05). Spheroid cells were more resistant to chemotherapeutic reagents than the congenetic adherent cells (P < 0.05). Flow cytometry showed enriched CD133+ population in sphere-forming cells (P < 0.05). CD133+ cells possessed high colony-formation ability than the CD133- population (P < 0.01). CD133+ cells injected into nude mice revealed higher tumorigenicity than their antigen-negative counterparts (P < 0.05).

CONCLUSION: CD133 may be a cell surface marker for CSCs in GBC.

CD133+ gallbladder carcinoma cells exhibit self-renewal ability and tumorigenicity

Organ-specific tumor cell adhesion to extracellular matrix (ECM) components and cell migration into host organs often involve integrin-mediated cellular processes. Direct integrin-mediated cell adhesion to ECM components in the space of Disse appears to be required for the successful liver metastatic formation of colon cancer. In the present study, human colon cancer HT-29 cells were transfected by liposome with integrin-β1 antisense oligodeoxynucleotide (ASODN). The integrin-β1 gene expression in HT-29 cells was significantly down-regulated. The migration of HT-29 cells was assayed using transwell cell culture chambers in vitro. The number of migrating HT-29 cells in experimental group was far less than that in control group (P<0.05). The models of hepatic metastasis in nude mice were established by the intrasplenic injection of transfected HT-29 cells. Thirty days later, the nude mice were killed and the average number of hepatic metastases (4.00±0.93 per mouse), average volume (10.10±6.50 mm3 per mouse), average weight (0.0440±0.0008 g per mouse) in experimental group were remarkably reduced as compared with those in control group (P<0.05). Integrin-β1 expression in the hepatic metastasis was studied by immunohistochemistry (SP). Positive cell percentage of hepatic metastases in experimental group was markedly decreased as compared with that in control group (P<0.05). It was concluded that integrin-β1 may take part in hepatic metastasis, and down-regulation of integrin-β1 expression may play a key role in decreasing migration and hepatic metastasis of human colon carcinoma cells (HT-29).

Effects of down-regulation of integrin-β1 expression on migration and hepatic metastasis of human colon carcinoma

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CD133+ gallbladder carcinoma cells exhibit self-renewal ability and tumorigenicity

Effects of down-regulation of integrin-β1 expression on migration and hepatic metastasis of human colon carcinoma