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Endosialin‑expressing bone sarcoma stem‑like cells are highly tumor‑initiating and invasive.

Sun DX, Liao GJ, Liu KG, Jian H - Mol Med Rep (2015)

Bottom Line: The present study demonstrated that primary human osteosarcoma samples contained a SP of about 3.9% which overexpressed ABC transporters, including ABCA1, ABCB1, ABCB2 and ABCG2, which are associated with drug resistance and may have contributed to multi‑drug resistance of SP cells.Furthermore, these SP cells displayed increased expression of endosialin (CD248) and other stem cell surface proteins, including CD133, octamer‑binding transcription factor 3/4A, Nanog and Nestin, which are ultimately responsible for high self‑renewal and deregulated cell proliferation.In addition, it was shown that endosialin‑overexpressing SP cells were able to regenerate the tumor population and had a high invasive potential.

View Article: PubMed Central - PubMed

Affiliation: Department of Operating Room, Yantaishan Hospital, Yantai, Shandong 264000, P.R. China.

ABSTRACT
It has been reported that the presence of a small group of cancer stem‑like 'side population (SP)' cells is responsible for therapy failure and tumor recurrence. The present study demonstrated that primary human osteosarcoma samples contained a SP of about 3.9% which overexpressed ABC transporters, including ABCA1, ABCB1, ABCB2 and ABCG2, which are associated with drug resistance and may have contributed to multi‑drug resistance of SP cells. Furthermore, these SP cells displayed increased expression of endosialin (CD248) and other stem cell surface proteins, including CD133, octamer‑binding transcription factor 3/4A, Nanog and Nestin, which are ultimately responsible for high self‑renewal and deregulated cell proliferation. In addition, it was shown that endosialin‑overexpressing SP cells were able to regenerate the tumor population and had a high invasive potential. Therefore, the present study suggested that osteosarcoma SP cells were cancer stem cells, as they displayed stem‑like properties; furthermore, endosialin may be a potential target to prevent osteosarcoma recurrence following chemotherapy.

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Related in: MedlinePlus

Comparison of gene and protein expression levels between SP and non-SP cells. (A) Western blots showing elevated expression of endosialin and stem cell surface proteins in SP cells. (B) Quantified results of reverse-transcription quantitative polymerase chain reaction analysis for expression of endosialin and other stemness genes in SP and non-SP cells. The elevated mRNA expression of endosialin, CD133, nestin, Oct-3/4A and Nanog are significantly higher in SP cells. The bar represents the standard deviation and **P<0.01 for non-SP vs. SP. SP, side population; ABC, adenosine triphosphate binding cassette; Oct, octamer-binding transcription factor.
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f2-mmr-12-04-5665: Comparison of gene and protein expression levels between SP and non-SP cells. (A) Western blots showing elevated expression of endosialin and stem cell surface proteins in SP cells. (B) Quantified results of reverse-transcription quantitative polymerase chain reaction analysis for expression of endosialin and other stemness genes in SP and non-SP cells. The elevated mRNA expression of endosialin, CD133, nestin, Oct-3/4A and Nanog are significantly higher in SP cells. The bar represents the standard deviation and **P<0.01 for non-SP vs. SP. SP, side population; ABC, adenosine triphosphate binding cassette; Oct, octamer-binding transcription factor.

Mentions: It was previously shown that in osteosarcoma cell lines, endosialin (CD248) was expressed in SP cells (18). Hence, in the present study, the osteosarcoma SP cells identified by FACS were validated for the expression of endosialin and other stem-cell surface proteins. As shown in Fig. 2A, endosialin expression was elevated in SP cells compared with that in non-SP cells. Similarly, stem cell surface proteins, including CD133, Oct-3/4A (one of the isoforms of Oct-4 proteins) and Nanog were markedly elevated in SP cells (Fig. 2A). The increased expression of the abovementioned stem cell proteins in SP cells were thought to be involved in the maintenance of self-renewal and the high proliferation rate of SP cells. Next, the present study investigated the transcriptional upregulation of these genes by RT-PCR. The relative mRNA expression of endosialin, Nestin, Oct-3/4A and Naong was significantly elevated in SP cells compared with that in non-SP cells (Fig. 2B). In addition, immunofluorescence microscopy revealed that the osteosarcoma SP cells showed positivity for CD133, Oct3/4A (punctate nuclear staining), endosialin and Nanog (punctate nuclear staining), whereas the non-SP cells showed either low or expression (Fig. 3).


Endosialin‑expressing bone sarcoma stem‑like cells are highly tumor‑initiating and invasive.

Sun DX, Liao GJ, Liu KG, Jian H - Mol Med Rep (2015)

Comparison of gene and protein expression levels between SP and non-SP cells. (A) Western blots showing elevated expression of endosialin and stem cell surface proteins in SP cells. (B) Quantified results of reverse-transcription quantitative polymerase chain reaction analysis for expression of endosialin and other stemness genes in SP and non-SP cells. The elevated mRNA expression of endosialin, CD133, nestin, Oct-3/4A and Nanog are significantly higher in SP cells. The bar represents the standard deviation and **P<0.01 for non-SP vs. SP. SP, side population; ABC, adenosine triphosphate binding cassette; Oct, octamer-binding transcription factor.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581793&req=5

f2-mmr-12-04-5665: Comparison of gene and protein expression levels between SP and non-SP cells. (A) Western blots showing elevated expression of endosialin and stem cell surface proteins in SP cells. (B) Quantified results of reverse-transcription quantitative polymerase chain reaction analysis for expression of endosialin and other stemness genes in SP and non-SP cells. The elevated mRNA expression of endosialin, CD133, nestin, Oct-3/4A and Nanog are significantly higher in SP cells. The bar represents the standard deviation and **P<0.01 for non-SP vs. SP. SP, side population; ABC, adenosine triphosphate binding cassette; Oct, octamer-binding transcription factor.
Mentions: It was previously shown that in osteosarcoma cell lines, endosialin (CD248) was expressed in SP cells (18). Hence, in the present study, the osteosarcoma SP cells identified by FACS were validated for the expression of endosialin and other stem-cell surface proteins. As shown in Fig. 2A, endosialin expression was elevated in SP cells compared with that in non-SP cells. Similarly, stem cell surface proteins, including CD133, Oct-3/4A (one of the isoforms of Oct-4 proteins) and Nanog were markedly elevated in SP cells (Fig. 2A). The increased expression of the abovementioned stem cell proteins in SP cells were thought to be involved in the maintenance of self-renewal and the high proliferation rate of SP cells. Next, the present study investigated the transcriptional upregulation of these genes by RT-PCR. The relative mRNA expression of endosialin, Nestin, Oct-3/4A and Naong was significantly elevated in SP cells compared with that in non-SP cells (Fig. 2B). In addition, immunofluorescence microscopy revealed that the osteosarcoma SP cells showed positivity for CD133, Oct3/4A (punctate nuclear staining), endosialin and Nanog (punctate nuclear staining), whereas the non-SP cells showed either low or expression (Fig. 3).

Bottom Line: The present study demonstrated that primary human osteosarcoma samples contained a SP of about 3.9% which overexpressed ABC transporters, including ABCA1, ABCB1, ABCB2 and ABCG2, which are associated with drug resistance and may have contributed to multi‑drug resistance of SP cells.Furthermore, these SP cells displayed increased expression of endosialin (CD248) and other stem cell surface proteins, including CD133, octamer‑binding transcription factor 3/4A, Nanog and Nestin, which are ultimately responsible for high self‑renewal and deregulated cell proliferation.In addition, it was shown that endosialin‑overexpressing SP cells were able to regenerate the tumor population and had a high invasive potential.

View Article: PubMed Central - PubMed

Affiliation: Department of Operating Room, Yantaishan Hospital, Yantai, Shandong 264000, P.R. China.

ABSTRACT
It has been reported that the presence of a small group of cancer stem‑like 'side population (SP)' cells is responsible for therapy failure and tumor recurrence. The present study demonstrated that primary human osteosarcoma samples contained a SP of about 3.9% which overexpressed ABC transporters, including ABCA1, ABCB1, ABCB2 and ABCG2, which are associated with drug resistance and may have contributed to multi‑drug resistance of SP cells. Furthermore, these SP cells displayed increased expression of endosialin (CD248) and other stem cell surface proteins, including CD133, octamer‑binding transcription factor 3/4A, Nanog and Nestin, which are ultimately responsible for high self‑renewal and deregulated cell proliferation. In addition, it was shown that endosialin‑overexpressing SP cells were able to regenerate the tumor population and had a high invasive potential. Therefore, the present study suggested that osteosarcoma SP cells were cancer stem cells, as they displayed stem‑like properties; furthermore, endosialin may be a potential target to prevent osteosarcoma recurrence following chemotherapy.

Show MeSH
Related in: MedlinePlus