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Endothelial cells promote stem-like phenotype of glioma cells through activating the Hedgehog pathway.

Yan GN, Yang L, Lv YF, Shi Y, Shen LL, Yao XH, Guo QN, Zhang P, Cui YH, Zhang X, Bian XW, Guo DY - J. Pathol. (2014)

Bottom Line: Microenvironmental regulation of cancer stem cells (CSCs) strongly influences the onset and spread of cancer.We observed that CD133(+) GSCs were located closely to Shh(+) endothelial cells in specimens of human glioblastoma multiforme (GBM).Knockdown of Smo in glioma cells led to a significant reduction of their CSC-like phenotype formation in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology and Southwest Cancer Centre, Southwest Hospital, Third Military Medical University and Key Laboratory of Tumour Immunopathology, Ministry of Education of China, Chongqing, 400038, People's Republic of China.

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Endothelial cells up-regulate expression of CSC-associated genes in glioma cells. As compared to GL261 alone, expression of GSC-associated genes Olig2, Bmi1 and Sox2 were elevated in GL261 cells co-cultured with b.END3 cells: (A) real-time RT–PCR, *p < 0.05; (B) western blot, tubulin was used as control. As compared to U251 cells alone, expressions of Olig2, Bmi1 and Sox2 were elevated in U251 cells co-cultured with HUVECs; (C) real-time RT–PCR, *p < 0.05; (D) western blot, tubulin was used as control. (E) Immunofluorescence staining revealed that expressions of Bmi1 (upper), Sox2 (middle) and Olig2 (lower) were increased in GL261 cells co-cultured with b.END3 cells. Right panels are partial enlargements of the corresponding left panels.
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fig03: Endothelial cells up-regulate expression of CSC-associated genes in glioma cells. As compared to GL261 alone, expression of GSC-associated genes Olig2, Bmi1 and Sox2 were elevated in GL261 cells co-cultured with b.END3 cells: (A) real-time RT–PCR, *p < 0.05; (B) western blot, tubulin was used as control. As compared to U251 cells alone, expressions of Olig2, Bmi1 and Sox2 were elevated in U251 cells co-cultured with HUVECs; (C) real-time RT–PCR, *p < 0.05; (D) western blot, tubulin was used as control. (E) Immunofluorescence staining revealed that expressions of Bmi1 (upper), Sox2 (middle) and Olig2 (lower) were increased in GL261 cells co-cultured with b.END3 cells. Right panels are partial enlargements of the corresponding left panels.

Mentions: Olig2, Sox2 and Bmi1, key genes in maintaining the stemness of GSCs 18,27,28, could be detected in both GL261 and U251 cells. As shown in Figure 3, Olig2, Bmi1 and Sox2 were all over-expressed in GL261 cells co-cultured with b.END3 cells, and the expression change of Olig2 was most obvious among them (Figure 3A), which was also in parallel with their corresponding protein levels, as demonstrated by western blot analysis (Figure 3B). In the U251 cells cultured with HUVECs, Olig2, Bmi1 and Sox2 were also over-expressed at the levels of both mRNA (Figure 3C) and protein (Figure 3D). Immunofluorescence confocal microscopy not only revealed increased intensities of Olig2, Bmi1 and Sox2 but also that Olig2 was translocated from the cytoplasm to the nuclei in GL261 cells after co-culture with ECs (Figure 3E). These data demonstrate that ECs are able to up-regulate the expression of stemness-related genes in glioma cells upon their interaction with each other.


Endothelial cells promote stem-like phenotype of glioma cells through activating the Hedgehog pathway.

Yan GN, Yang L, Lv YF, Shi Y, Shen LL, Yao XH, Guo QN, Zhang P, Cui YH, Zhang X, Bian XW, Guo DY - J. Pathol. (2014)

Endothelial cells up-regulate expression of CSC-associated genes in glioma cells. As compared to GL261 alone, expression of GSC-associated genes Olig2, Bmi1 and Sox2 were elevated in GL261 cells co-cultured with b.END3 cells: (A) real-time RT–PCR, *p < 0.05; (B) western blot, tubulin was used as control. As compared to U251 cells alone, expressions of Olig2, Bmi1 and Sox2 were elevated in U251 cells co-cultured with HUVECs; (C) real-time RT–PCR, *p < 0.05; (D) western blot, tubulin was used as control. (E) Immunofluorescence staining revealed that expressions of Bmi1 (upper), Sox2 (middle) and Olig2 (lower) were increased in GL261 cells co-cultured with b.END3 cells. Right panels are partial enlargements of the corresponding left panels.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260128&req=5

fig03: Endothelial cells up-regulate expression of CSC-associated genes in glioma cells. As compared to GL261 alone, expression of GSC-associated genes Olig2, Bmi1 and Sox2 were elevated in GL261 cells co-cultured with b.END3 cells: (A) real-time RT–PCR, *p < 0.05; (B) western blot, tubulin was used as control. As compared to U251 cells alone, expressions of Olig2, Bmi1 and Sox2 were elevated in U251 cells co-cultured with HUVECs; (C) real-time RT–PCR, *p < 0.05; (D) western blot, tubulin was used as control. (E) Immunofluorescence staining revealed that expressions of Bmi1 (upper), Sox2 (middle) and Olig2 (lower) were increased in GL261 cells co-cultured with b.END3 cells. Right panels are partial enlargements of the corresponding left panels.
Mentions: Olig2, Sox2 and Bmi1, key genes in maintaining the stemness of GSCs 18,27,28, could be detected in both GL261 and U251 cells. As shown in Figure 3, Olig2, Bmi1 and Sox2 were all over-expressed in GL261 cells co-cultured with b.END3 cells, and the expression change of Olig2 was most obvious among them (Figure 3A), which was also in parallel with their corresponding protein levels, as demonstrated by western blot analysis (Figure 3B). In the U251 cells cultured with HUVECs, Olig2, Bmi1 and Sox2 were also over-expressed at the levels of both mRNA (Figure 3C) and protein (Figure 3D). Immunofluorescence confocal microscopy not only revealed increased intensities of Olig2, Bmi1 and Sox2 but also that Olig2 was translocated from the cytoplasm to the nuclei in GL261 cells after co-culture with ECs (Figure 3E). These data demonstrate that ECs are able to up-regulate the expression of stemness-related genes in glioma cells upon their interaction with each other.

Bottom Line: Microenvironmental regulation of cancer stem cells (CSCs) strongly influences the onset and spread of cancer.We observed that CD133(+) GSCs were located closely to Shh(+) endothelial cells in specimens of human glioblastoma multiforme (GBM).Knockdown of Smo in glioma cells led to a significant reduction of their CSC-like phenotype formation in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology and Southwest Cancer Centre, Southwest Hospital, Third Military Medical University and Key Laboratory of Tumour Immunopathology, Ministry of Education of China, Chongqing, 400038, People's Republic of China.

Show MeSH
Related in: MedlinePlus