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Potassium channel ether à go-go1 is aberrantly expressed in human liposarcoma and promotes tumorigenesis.

Wu J, Zhong D, Wei Y, Wu X, Kang L, Ding Z - Biomed Res Int (2014)

Bottom Line: The ether à go-go1 (Eag1) channel is overexpressed in a variety of cancers.Then, the protein expression of Eag1 in 131 different adipose tissues from 109 patients was detected by immunohistochemistry.It was found that Eag1 was aberrantly expressed in over 67% liposarcomas, with a higher frequency than in lipoma, hyperplasia, inflammation, and normal adipose tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, the Affiliated Southeast Hospital of Xiamen University, Orthopaedic Center of People's Liberation Army, Zhanghua Road 269, Zhangzhou 363000, China.

ABSTRACT
The ether à go-go1 (Eag1) channel is overexpressed in a variety of cancers. However, the expression and function of Eag1 in liposarcoma are poorly understood. In the present study, the mRNA expression of Eag1 in different adipose tissue samples was examined by real-time PCR. Then, the protein expression of Eag1 in 131 different adipose tissues from 109 patients was detected by immunohistochemistry. Next, the associations between Eag1 expression and clinicopathological features of liposarcoma were analyzed. In addition, the effects of Eag1 on liposarcoma cell proliferation and cycle were evaluated by CCK-8, colony formation, xenograft mouse model, and flow cytometry, respectively. Finally, the activation of p38 mitogen-activated protein kinase (MAPK) was detected by Western blot analysis to explain the detailed mechanisms of oncogenic potential of Eag1 in liposarcoma. It was found that Eag1 was aberrantly expressed in over 67% liposarcomas, with a higher frequency than in lipoma, hyperplasia, inflammation, and normal adipose tissues. However, Eag1 expression was not correlated with clinicopathological features of liposarcoma. Eag1 inhibitor imipramine or Eag1-shRNA significantly suppressed the proliferation of liposarcoma cells in vitro and in vivo, accompanying with accumulation of cells in the G1 phase. These results suggest that Eag1 plays an important role in regulating the proliferation and cell cycle of liposarcoma cells and might be a potential therapeutic target for liposarcoma.

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The expression of Eag1 in different adipose tissues diseases samples. (a) Real-time PCR reveals increased Eag1 expression in liposarcoma. The mRNA expression level is normalized with GAPDH and the value in normal adipose tissues is set as 1 for the calibration. ∗∗∗P < 0.001. (b) Eag1 immunostaining in a human brain specimen (positive control, (A)), 67.5% liposarcoma (B), 42.1% lipoma (C), 40.0% hyperplasia of adipose tissues (D), 25.0% panniculitis (E), and no positive Eag1 staining in 9 normal adipose tissues (F) and 3 skeletal muscle tissues (negative control, (G)). Images are captured using an OLYMPUS light microscope equipped with a CCD color camera at a 200x magnification.
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fig1: The expression of Eag1 in different adipose tissues diseases samples. (a) Real-time PCR reveals increased Eag1 expression in liposarcoma. The mRNA expression level is normalized with GAPDH and the value in normal adipose tissues is set as 1 for the calibration. ∗∗∗P < 0.001. (b) Eag1 immunostaining in a human brain specimen (positive control, (A)), 67.5% liposarcoma (B), 42.1% lipoma (C), 40.0% hyperplasia of adipose tissues (D), 25.0% panniculitis (E), and no positive Eag1 staining in 9 normal adipose tissues (F) and 3 skeletal muscle tissues (negative control, (G)). Images are captured using an OLYMPUS light microscope equipped with a CCD color camera at a 200x magnification.

Mentions: First, we compared the mRNA expression level of Eag1 in different adipose tissue samples by real-time PCR. There was a clear increase in Eag1 mRNA level in liposarcoma samples (n = 3), compared to other tissues (n = 3) (Figure 1(a)). Next, Eag1 polyclonal antibody was employed for immunohistochemical staining. Positive Eag1 staining was detected in 54/80 (67.5%) liposarcoma, 8/19 (42.1%) lipoma, 6/15 (40.0%) hyperplasia of adipose tissues, and 2/8 panniculitis (25.0%), but negative Eag1 staining was detected in 9 normal adipose tissues (Table 1). The positive signal was detected mainly in the cytoplasm, consistent with previous reports [11, 16]. As a positive control Eag1 staining was detected in a normal human brain sample, negative staining for Eag1 was observed in a skeletal muscle tissue. The representative images of Eag1 staining are shown in Figure 1(b). Statistical analysis indicated that positive expression of Eag1 in human liposarcoma was significantly different from that in other adipose tissues diseases (P < 0.05, Table 1).


Potassium channel ether à go-go1 is aberrantly expressed in human liposarcoma and promotes tumorigenesis.

Wu J, Zhong D, Wei Y, Wu X, Kang L, Ding Z - Biomed Res Int (2014)

The expression of Eag1 in different adipose tissues diseases samples. (a) Real-time PCR reveals increased Eag1 expression in liposarcoma. The mRNA expression level is normalized with GAPDH and the value in normal adipose tissues is set as 1 for the calibration. ∗∗∗P < 0.001. (b) Eag1 immunostaining in a human brain specimen (positive control, (A)), 67.5% liposarcoma (B), 42.1% lipoma (C), 40.0% hyperplasia of adipose tissues (D), 25.0% panniculitis (E), and no positive Eag1 staining in 9 normal adipose tissues (F) and 3 skeletal muscle tissues (negative control, (G)). Images are captured using an OLYMPUS light microscope equipped with a CCD color camera at a 200x magnification.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4127296&req=5

fig1: The expression of Eag1 in different adipose tissues diseases samples. (a) Real-time PCR reveals increased Eag1 expression in liposarcoma. The mRNA expression level is normalized with GAPDH and the value in normal adipose tissues is set as 1 for the calibration. ∗∗∗P < 0.001. (b) Eag1 immunostaining in a human brain specimen (positive control, (A)), 67.5% liposarcoma (B), 42.1% lipoma (C), 40.0% hyperplasia of adipose tissues (D), 25.0% panniculitis (E), and no positive Eag1 staining in 9 normal adipose tissues (F) and 3 skeletal muscle tissues (negative control, (G)). Images are captured using an OLYMPUS light microscope equipped with a CCD color camera at a 200x magnification.
Mentions: First, we compared the mRNA expression level of Eag1 in different adipose tissue samples by real-time PCR. There was a clear increase in Eag1 mRNA level in liposarcoma samples (n = 3), compared to other tissues (n = 3) (Figure 1(a)). Next, Eag1 polyclonal antibody was employed for immunohistochemical staining. Positive Eag1 staining was detected in 54/80 (67.5%) liposarcoma, 8/19 (42.1%) lipoma, 6/15 (40.0%) hyperplasia of adipose tissues, and 2/8 panniculitis (25.0%), but negative Eag1 staining was detected in 9 normal adipose tissues (Table 1). The positive signal was detected mainly in the cytoplasm, consistent with previous reports [11, 16]. As a positive control Eag1 staining was detected in a normal human brain sample, negative staining for Eag1 was observed in a skeletal muscle tissue. The representative images of Eag1 staining are shown in Figure 1(b). Statistical analysis indicated that positive expression of Eag1 in human liposarcoma was significantly different from that in other adipose tissues diseases (P < 0.05, Table 1).

Bottom Line: The ether à go-go1 (Eag1) channel is overexpressed in a variety of cancers.Then, the protein expression of Eag1 in 131 different adipose tissues from 109 patients was detected by immunohistochemistry.It was found that Eag1 was aberrantly expressed in over 67% liposarcomas, with a higher frequency than in lipoma, hyperplasia, inflammation, and normal adipose tissues.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, the Affiliated Southeast Hospital of Xiamen University, Orthopaedic Center of People's Liberation Army, Zhanghua Road 269, Zhangzhou 363000, China.

ABSTRACT
The ether à go-go1 (Eag1) channel is overexpressed in a variety of cancers. However, the expression and function of Eag1 in liposarcoma are poorly understood. In the present study, the mRNA expression of Eag1 in different adipose tissue samples was examined by real-time PCR. Then, the protein expression of Eag1 in 131 different adipose tissues from 109 patients was detected by immunohistochemistry. Next, the associations between Eag1 expression and clinicopathological features of liposarcoma were analyzed. In addition, the effects of Eag1 on liposarcoma cell proliferation and cycle were evaluated by CCK-8, colony formation, xenograft mouse model, and flow cytometry, respectively. Finally, the activation of p38 mitogen-activated protein kinase (MAPK) was detected by Western blot analysis to explain the detailed mechanisms of oncogenic potential of Eag1 in liposarcoma. It was found that Eag1 was aberrantly expressed in over 67% liposarcomas, with a higher frequency than in lipoma, hyperplasia, inflammation, and normal adipose tissues. However, Eag1 expression was not correlated with clinicopathological features of liposarcoma. Eag1 inhibitor imipramine or Eag1-shRNA significantly suppressed the proliferation of liposarcoma cells in vitro and in vivo, accompanying with accumulation of cells in the G1 phase. These results suggest that Eag1 plays an important role in regulating the proliferation and cell cycle of liposarcoma cells and might be a potential therapeutic target for liposarcoma.

Show MeSH
Related in: MedlinePlus