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

Evaluation of multi-differentiation potential of T3A-A3 cells. The multi-differentiation potential of T3A-A3 cells was analyzed after induction with melanoma, lymphoma and prostate cancer cell- or tissue-derived conditioned culture medium. The morphological changes of T3A-A3 after induction were photographed. The tumor-specific markers were compared between T3A-A3 cells before and after induction by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo. (a) Morphological changes and expression of gp100 in T3A-A3 after induction by melanoma-derived conditioned culture medium. (b) Morphological changes and expression of CD10 in T3A-A3 after induction by lymphoma-derived conditioned culture medium. (c) Morphological changes and expression of PSA in T3A-A3 after induction by prostate cancer-derived conditioned culture medium. (1: positive control cells (corresponding cancer cell line); 2: T3A-A3 cells; 3: T3A-A3 cells after induction with tumor cell-derived conditioned culture medium; 4: T3A-A3 cells induced with tumor tissue-derived conditioned culture medium)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Evaluation of multi-differentiation potential of T3A-A3 cells. The multi-differentiation potential of T3A-A3 cells was analyzed after induction with melanoma, lymphoma and prostate cancer cell- or tissue-derived conditioned culture medium. The morphological changes of T3A-A3 after induction were photographed. The tumor-specific markers were compared between T3A-A3 cells before and after induction by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo. (a) Morphological changes and expression of gp100 in T3A-A3 after induction by melanoma-derived conditioned culture medium. (b) Morphological changes and expression of CD10 in T3A-A3 after induction by lymphoma-derived conditioned culture medium. (c) Morphological changes and expression of PSA in T3A-A3 after induction by prostate cancer-derived conditioned culture medium. (1: positive control cells (corresponding cancer cell line); 2: T3A-A3 cells; 3: T3A-A3 cells after induction with tumor cell-derived conditioned culture medium; 4: T3A-A3 cells induced with tumor tissue-derived conditioned culture medium)

Mentions: Phase-contrast microscopy analysis revealed changes in the cell morphology of the T3A-A3 cells following melanoma cell-directed differentiation after 72 h induction, from polygonal-like into longitudinal stretched cells, which are similar to melanoma cell line SK-MEL-1, especially in the group of tissue-conditioned medium (Figure 4a). Furthermore, melanoma-specific expressing marker gp100 detected by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo, respectively at the gene, protein and cell levels became significantly positive in T3A-A3 cells after induction, but T3A-A3 cells did not express gp100 before induction.


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)

Evaluation of multi-differentiation potential of T3A-A3 cells. The multi-differentiation potential of T3A-A3 cells was analyzed after induction with melanoma, lymphoma and prostate cancer cell- or tissue-derived conditioned culture medium. The morphological changes of T3A-A3 after induction were photographed. The tumor-specific markers were compared between T3A-A3 cells before and after induction by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo. (a) Morphological changes and expression of gp100 in T3A-A3 after induction by melanoma-derived conditioned culture medium. (b) Morphological changes and expression of CD10 in T3A-A3 after induction by lymphoma-derived conditioned culture medium. (c) Morphological changes and expression of PSA in T3A-A3 after induction by prostate cancer-derived conditioned culture medium. (1: positive control cells (corresponding cancer cell line); 2: T3A-A3 cells; 3: T3A-A3 cells after induction with tumor cell-derived conditioned culture medium; 4: T3A-A3 cells induced with tumor tissue-derived conditioned culture medium)
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig4: Evaluation of multi-differentiation potential of T3A-A3 cells. The multi-differentiation potential of T3A-A3 cells was analyzed after induction with melanoma, lymphoma and prostate cancer cell- or tissue-derived conditioned culture medium. The morphological changes of T3A-A3 after induction were photographed. The tumor-specific markers were compared between T3A-A3 cells before and after induction by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo. (a) Morphological changes and expression of gp100 in T3A-A3 after induction by melanoma-derived conditioned culture medium. (b) Morphological changes and expression of CD10 in T3A-A3 after induction by lymphoma-derived conditioned culture medium. (c) Morphological changes and expression of PSA in T3A-A3 after induction by prostate cancer-derived conditioned culture medium. (1: positive control cells (corresponding cancer cell line); 2: T3A-A3 cells; 3: T3A-A3 cells after induction with tumor cell-derived conditioned culture medium; 4: T3A-A3 cells induced with tumor tissue-derived conditioned culture medium)
Mentions: Phase-contrast microscopy analysis revealed changes in the cell morphology of the T3A-A3 cells following melanoma cell-directed differentiation after 72 h induction, from polygonal-like into longitudinal stretched cells, which are similar to melanoma cell line SK-MEL-1, especially in the group of tissue-conditioned medium (Figure 4a). Furthermore, melanoma-specific expressing marker gp100 detected by RT-PCR, western blot, immunofluorescence staining in vitro and immunohistochemistry staining on graft tumor tissue in vivo, respectively at the gene, protein and cell levels became significantly positive in T3A-A3 cells after induction, but T3A-A3 cells did not express gp100 before induction.

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