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Apurinic/apyrimidinic endonuclease 1 regulates angiogenesis in a transforming growth factor β-dependent manner in human osteosarcoma.

Jiang X, Shan J, Dai N, Zhong Z, Qing Y, Yang Y, Zhang S, Li C, Sui J, Ren T, Li M, Wang D - Cancer Sci. (2015)

Bottom Line: In addition, APE1-siRNA led to suppression of angiogenesis in vitro based on HUVECs in Transwell and Matrigel tube formation assays.Reduced secretory protein level of TGFβ of culture medium also resulted in decreased phosphorylation of Smad3 of HUVECs.In a mouse xenograft model, siRNA-mediated silencing of APE1 downregulated TGFβ expression, tumor size, and MVD.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center, Daping Hospital and Research Institute of Surgery, Third Military Medical University, Chongqing, China.

No MeSH data available.


Related in: MedlinePlus

Osteosarcoma 9901 cells transfected with two apurinic/apyrimidinic endonuclease 1 (APE1)-siRNAs, negative control (scramble), or Optimum I as blank (control). (a) Western blot images. (b,c) Western blot analysis shows that APE1-siRNA suppresses expression of both APE1 and transforming growth factor β1 (TGFβ1). (d) ELISA assays show the concentration of TGFβ1 in the supernatant of the control, scramble, and treatment groups. **P < 0.01.
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fig03: Osteosarcoma 9901 cells transfected with two apurinic/apyrimidinic endonuclease 1 (APE1)-siRNAs, negative control (scramble), or Optimum I as blank (control). (a) Western blot images. (b,c) Western blot analysis shows that APE1-siRNA suppresses expression of both APE1 and transforming growth factor β1 (TGFβ1). (d) ELISA assays show the concentration of TGFβ1 in the supernatant of the control, scramble, and treatment groups. **P < 0.01.

Mentions: We then tested whether TGFβ1 expression is affected by APE1 exogenous manipulation in 9901 cells by Western blot. As shown in Figure 3, the expression of APE1 was decreased in APE1-siRNA transfected cells when compared with the scramble-siRNA transfected cells and blank control. When APE1 was knocked down, TGFβ1 expression was downregulated. The percentage inhibition of APE1 and TGFβ1 protein was 88.75% and 63.1% compared to the blank control, respectively (P < 0.01), and there were no differences between the scramble and blank control. To evaluate the secretory level of TGFβ1 affected by APE1 deficiency, an ELISA was carried out. The results indicated that knocking down APE1 in 9901 cells results in a significant decrease in TGFβ1 levels in the supernatant when compared with control groups (435.23 ± 22.80 pg/mL for the APE1-siRNA group, 731.610 ± 48.949 pg/mL for the blank control group, and 755.75 ± 51.36 pg/mL for the scramble group with an inhibition level of 40.51% (Fig. 3d). To investigate the mRNA level of TGFβ1 expression after knocking down APE1-siRNA, we carried out an RT-PCR assay (P < 0.01; Fig. 4a). In APE1-siRNA transfected 9901 cells, the expression of APE1 was decreased by 50.81% (P < 0.01; Fig. 4b). As a result of APE1 knockdown, TGFβ1 expression was downregulated by 49.54% (P < 0.01; Fig. 4c). The above experiments confirmed that exogenous manipulation of APE1 results in altered expression of TGFβ1, suggesting a regulatory role of APE1 in TGFβ1 signaling.


Apurinic/apyrimidinic endonuclease 1 regulates angiogenesis in a transforming growth factor β-dependent manner in human osteosarcoma.

Jiang X, Shan J, Dai N, Zhong Z, Qing Y, Yang Y, Zhang S, Li C, Sui J, Ren T, Li M, Wang D - Cancer Sci. (2015)

Osteosarcoma 9901 cells transfected with two apurinic/apyrimidinic endonuclease 1 (APE1)-siRNAs, negative control (scramble), or Optimum I as blank (control). (a) Western blot images. (b,c) Western blot analysis shows that APE1-siRNA suppresses expression of both APE1 and transforming growth factor β1 (TGFβ1). (d) ELISA assays show the concentration of TGFβ1 in the supernatant of the control, scramble, and treatment groups. **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4638009&req=5

fig03: Osteosarcoma 9901 cells transfected with two apurinic/apyrimidinic endonuclease 1 (APE1)-siRNAs, negative control (scramble), or Optimum I as blank (control). (a) Western blot images. (b,c) Western blot analysis shows that APE1-siRNA suppresses expression of both APE1 and transforming growth factor β1 (TGFβ1). (d) ELISA assays show the concentration of TGFβ1 in the supernatant of the control, scramble, and treatment groups. **P < 0.01.
Mentions: We then tested whether TGFβ1 expression is affected by APE1 exogenous manipulation in 9901 cells by Western blot. As shown in Figure 3, the expression of APE1 was decreased in APE1-siRNA transfected cells when compared with the scramble-siRNA transfected cells and blank control. When APE1 was knocked down, TGFβ1 expression was downregulated. The percentage inhibition of APE1 and TGFβ1 protein was 88.75% and 63.1% compared to the blank control, respectively (P < 0.01), and there were no differences between the scramble and blank control. To evaluate the secretory level of TGFβ1 affected by APE1 deficiency, an ELISA was carried out. The results indicated that knocking down APE1 in 9901 cells results in a significant decrease in TGFβ1 levels in the supernatant when compared with control groups (435.23 ± 22.80 pg/mL for the APE1-siRNA group, 731.610 ± 48.949 pg/mL for the blank control group, and 755.75 ± 51.36 pg/mL for the scramble group with an inhibition level of 40.51% (Fig. 3d). To investigate the mRNA level of TGFβ1 expression after knocking down APE1-siRNA, we carried out an RT-PCR assay (P < 0.01; Fig. 4a). In APE1-siRNA transfected 9901 cells, the expression of APE1 was decreased by 50.81% (P < 0.01; Fig. 4b). As a result of APE1 knockdown, TGFβ1 expression was downregulated by 49.54% (P < 0.01; Fig. 4c). The above experiments confirmed that exogenous manipulation of APE1 results in altered expression of TGFβ1, suggesting a regulatory role of APE1 in TGFβ1 signaling.

Bottom Line: In addition, APE1-siRNA led to suppression of angiogenesis in vitro based on HUVECs in Transwell and Matrigel tube formation assays.Reduced secretory protein level of TGFβ of culture medium also resulted in decreased phosphorylation of Smad3 of HUVECs.In a mouse xenograft model, siRNA-mediated silencing of APE1 downregulated TGFβ expression, tumor size, and MVD.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center, Daping Hospital and Research Institute of Surgery, Third Military Medical University, Chongqing, China.

No MeSH data available.


Related in: MedlinePlus