Limits...
Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells.

Liu H, Zhang W, Jia Y, Yu Q, Grau GE, Peng L, Ran Y, Yang Z, Deng H, Lou J - Cell Death Dis (2013)

Bottom Line: Here, we show that single-cell-cloned CSCs, accidentally obtained from a human liver cancer microvascular endothelial cells, express classic stem cell markers, genes associated with self-renewal and pluripotent factors and possess colony-forming ability in vitro and the ability of serial transplantation in vivo.Interestingly, this multilineage differentiation potential of single-cell-cloned liver CSCs sharply declined after the specific knockdown of octamer-binding transcription factor 4 (Oct4) alone, even though they were under the same induction conditions (carcinoma microenvironments).Simultaneously, Oct4 in CSCs is indispensable in this process.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, PR China.

ABSTRACT
Cancer stem cells (CSCs) are believed to be a promising target for cancer therapy because these cells are responsible for tumor development, maintenance and chemotherapy resistance. Finding out the critical factors regulating CSC fate is the key for target therapy of CSCs. Just as normal stem cells are regulated by their microenvironment (niche), CSCs are also regulated by cells in the tumor microenvironment. However, whether various tumor microenvironments can induce CSCs to differentiate into different cancer cells is not clear. Here, we show that single-cell-cloned CSCs, accidentally obtained from a human liver cancer microvascular endothelial cells, express classic stem cell markers, genes associated with self-renewal and pluripotent factors and possess colony-forming ability in vitro and the ability of serial transplantation in vivo. The single-cell-cloned CSCs treated with the different tumor cell/tissue-derived conditioned culture medium, which is a mimic of carcinoma microenvironment, could differentiate into corresponding tumor cells and express specific markers of the respective type of tumor cells at the gene, protein and cell levels, respectively. Interestingly, this multilineage differentiation potential of single-cell-cloned liver CSCs sharply declined after the specific knockdown of octamer-binding transcription factor 4 (Oct4) alone, even though they were under the same induction conditions (carcinoma microenvironments). These data support the hypothesis that single-cell-cloned liver CSCs have the potential of differentiating into different types of tumor cells, and the tumor microenvironment does play a crucial role in deciding differentiation directions. Simultaneously, Oct4 in CSCs is indispensable in this process. These factors are promising targets for liver CSC-specific therapy.

Show MeSH

Related in: MedlinePlus

Identification of T3A-A3 cells. (a) RT-PCR analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (b) Flow cytometric analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (c) Evaluation of the self-renewing capacity of T3A-A3 cells. Secondary colony formation ability (the first and second panels). Tumor sphere-forming ability (the middle panel). Histopathology of the primary and the secondary grafted tumor (the last two panels). (d) Evaluation of tumor properties of T3A-A3 cells. Comparison of chromosomal karyotype between human fetal liver cells, human liver cancer cells and T3A-A3 cells (upper panels). Evaluation of tumorigenic and metastatic capacities of T3A-A3 cells in SCID mice (bottom panels)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3824650&req=5

fig3: Identification of T3A-A3 cells. (a) RT-PCR analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (b) Flow cytometric analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (c) Evaluation of the self-renewing capacity of T3A-A3 cells. Secondary colony formation ability (the first and second panels). Tumor sphere-forming ability (the middle panel). Histopathology of the primary and the secondary grafted tumor (the last two panels). (d) Evaluation of tumor properties of T3A-A3 cells. Comparison of chromosomal karyotype between human fetal liver cells, human liver cancer cells and T3A-A3 cells (upper panels). Evaluation of tumorigenic and metastatic capacities of T3A-A3 cells in SCID mice (bottom panels)

Mentions: To further identify the cancer stem-like cell identity of T3A-A3, we detected the expressions of many vital genes and surface markers related to stemness. Using semiquantitative RT-PCR analyses (Figure 3a), we found that single-cell-cloned T3A-A3 cells express a wide variety of ‘stemness' genes such as CD133, SCF, CD117/c-kit, Nestin, CD34 and ABCG216, 17, 18 in different degrees. Moreover, T3A-A3 cells also express those genes associated with proliferation, self-renewal and differentiation of stem cells, such as Wnt/β-catenin, Notch-1, Hedgehog/SMO, Bmi-1 and Oct-4 pathways.19, 20, 21 Additionally, the T3A-A3 cells are positive for pluripotent factors related to induction of induced pluripotent stem (iPS) cells, including Oct-4, Klf4, C-myc, Sox2, Nanog and Lin28.22, 23 In this assay, human embryonic stem cell line (hESC) and human hepatoma cell line (BEL7402) were used as controls. All these markers in T3A-A3 cells were further confirmed by flow cytometry (Figure 3b).


Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells.

Liu H, Zhang W, Jia Y, Yu Q, Grau GE, Peng L, Ran Y, Yang Z, Deng H, Lou J - Cell Death Dis (2013)

Identification of T3A-A3 cells. (a) RT-PCR analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (b) Flow cytometric analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (c) Evaluation of the self-renewing capacity of T3A-A3 cells. Secondary colony formation ability (the first and second panels). Tumor sphere-forming ability (the middle panel). Histopathology of the primary and the secondary grafted tumor (the last two panels). (d) Evaluation of tumor properties of T3A-A3 cells. Comparison of chromosomal karyotype between human fetal liver cells, human liver cancer cells and T3A-A3 cells (upper panels). Evaluation of tumorigenic and metastatic capacities of T3A-A3 cells in SCID mice (bottom panels)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3824650&req=5

fig3: Identification of T3A-A3 cells. (a) RT-PCR analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (b) Flow cytometric analysis for the expressions of classic stem cell markers and genes associated with the proliferation and self-renewal of stem cells. (c) Evaluation of the self-renewing capacity of T3A-A3 cells. Secondary colony formation ability (the first and second panels). Tumor sphere-forming ability (the middle panel). Histopathology of the primary and the secondary grafted tumor (the last two panels). (d) Evaluation of tumor properties of T3A-A3 cells. Comparison of chromosomal karyotype between human fetal liver cells, human liver cancer cells and T3A-A3 cells (upper panels). Evaluation of tumorigenic and metastatic capacities of T3A-A3 cells in SCID mice (bottom panels)
Mentions: To further identify the cancer stem-like cell identity of T3A-A3, we detected the expressions of many vital genes and surface markers related to stemness. Using semiquantitative RT-PCR analyses (Figure 3a), we found that single-cell-cloned T3A-A3 cells express a wide variety of ‘stemness' genes such as CD133, SCF, CD117/c-kit, Nestin, CD34 and ABCG216, 17, 18 in different degrees. Moreover, T3A-A3 cells also express those genes associated with proliferation, self-renewal and differentiation of stem cells, such as Wnt/β-catenin, Notch-1, Hedgehog/SMO, Bmi-1 and Oct-4 pathways.19, 20, 21 Additionally, the T3A-A3 cells are positive for pluripotent factors related to induction of induced pluripotent stem (iPS) cells, including Oct-4, Klf4, C-myc, Sox2, Nanog and Lin28.22, 23 In this assay, human embryonic stem cell line (hESC) and human hepatoma cell line (BEL7402) were used as controls. All these markers in T3A-A3 cells were further confirmed by flow cytometry (Figure 3b).

Bottom Line: Here, we show that single-cell-cloned CSCs, accidentally obtained from a human liver cancer microvascular endothelial cells, express classic stem cell markers, genes associated with self-renewal and pluripotent factors and possess colony-forming ability in vitro and the ability of serial transplantation in vivo.Interestingly, this multilineage differentiation potential of single-cell-cloned liver CSCs sharply declined after the specific knockdown of octamer-binding transcription factor 4 (Oct4) alone, even though they were under the same induction conditions (carcinoma microenvironments).Simultaneously, Oct4 in CSCs is indispensable in this process.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, PR China.

ABSTRACT
Cancer stem cells (CSCs) are believed to be a promising target for cancer therapy because these cells are responsible for tumor development, maintenance and chemotherapy resistance. Finding out the critical factors regulating CSC fate is the key for target therapy of CSCs. Just as normal stem cells are regulated by their microenvironment (niche), CSCs are also regulated by cells in the tumor microenvironment. However, whether various tumor microenvironments can induce CSCs to differentiate into different cancer cells is not clear. Here, we show that single-cell-cloned CSCs, accidentally obtained from a human liver cancer microvascular endothelial cells, express classic stem cell markers, genes associated with self-renewal and pluripotent factors and possess colony-forming ability in vitro and the ability of serial transplantation in vivo. The single-cell-cloned CSCs treated with the different tumor cell/tissue-derived conditioned culture medium, which is a mimic of carcinoma microenvironment, could differentiate into corresponding tumor cells and express specific markers of the respective type of tumor cells at the gene, protein and cell levels, respectively. Interestingly, this multilineage differentiation potential of single-cell-cloned liver CSCs sharply declined after the specific knockdown of octamer-binding transcription factor 4 (Oct4) alone, even though they were under the same induction conditions (carcinoma microenvironments). These data support the hypothesis that single-cell-cloned liver CSCs have the potential of differentiating into different types of tumor cells, and the tumor microenvironment does play a crucial role in deciding differentiation directions. Simultaneously, Oct4 in CSCs is indispensable in this process. These factors are promising targets for liver CSC-specific therapy.

Show MeSH
Related in: MedlinePlus