Limits...
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.

Show MeSH

Related in: MedlinePlus

Eag1 blockage inhibits the proliferation and tumorigenicity of liposarcoma cells. (a) Western blot analysis of Eag1 protein levels in liposarcoma cells treated with Eag1 shRNA. Densitometric analysis of the bolts with GAPDH as loading control. The results are expressed as mean ± SD (n = 3). (b) The proliferation of liposarcoma cells is determined by CCK-8 assay after treatment with imipramine or Eag1 shRNA. The proliferation of liposarcoma cells is significantly reduced after treatment with imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 6). (c) The tumorigenicity of liposarcoma cells is determined by colony formation assay. The tumorigenicity of liposarcoma cells is significantly reduced after treatment with 20 μM imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 3). ∗∗P < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4127296&req=5

fig3: Eag1 blockage inhibits the proliferation and tumorigenicity of liposarcoma cells. (a) Western blot analysis of Eag1 protein levels in liposarcoma cells treated with Eag1 shRNA. Densitometric analysis of the bolts with GAPDH as loading control. The results are expressed as mean ± SD (n = 3). (b) The proliferation of liposarcoma cells is determined by CCK-8 assay after treatment with imipramine or Eag1 shRNA. The proliferation of liposarcoma cells is significantly reduced after treatment with imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 6). (c) The tumorigenicity of liposarcoma cells is determined by colony formation assay. The tumorigenicity of liposarcoma cells is significantly reduced after treatment with 20 μM imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 3). ∗∗P < 0.01.

Mentions: To characterize the oncogenic role of Eag1 in liposarcoma, we first inhibited Eag1 expression by Eag1 shRNA. As shown in Figure 3(a), Eag1 protein expression was effectively suppressed by Ad5-Eag1-shRNA. Next the effect of Eag1 knockdown on cell proliferation was determined by CCK-8 assay. The results revealed that the proliferation of SW-872 and 93T449 cells was inhibited by 39% and 31%, respectively, after Ad5-Eag1-shRNA infection. To confirm the oncogenic role of Eag1, we treated liposarcoma cells with imipramine, a nonspecific blocker of Eag1 activity. The results showed that the proliferation of SW-872 and 93T449 cells was inhibited by 28% and 22%, respectively, after the treatment with 20 μM imipramine (Figure 3(b)). Similar results were obtained from the colony formation experiment (Figure 3(c)). Collectively, these results suggest that Eag1 promotes the proliferation of liposarcoma cells in vitro.


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)

Eag1 blockage inhibits the proliferation and tumorigenicity of liposarcoma cells. (a) Western blot analysis of Eag1 protein levels in liposarcoma cells treated with Eag1 shRNA. Densitometric analysis of the bolts with GAPDH as loading control. The results are expressed as mean ± SD (n = 3). (b) The proliferation of liposarcoma cells is determined by CCK-8 assay after treatment with imipramine or Eag1 shRNA. The proliferation of liposarcoma cells is significantly reduced after treatment with imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 6). (c) The tumorigenicity of liposarcoma cells is determined by colony formation assay. The tumorigenicity of liposarcoma cells is significantly reduced after treatment with 20 μM imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 3). ∗∗P < 0.01.
© Copyright Policy
Related In: Results  -  Collection

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

fig3: Eag1 blockage inhibits the proliferation and tumorigenicity of liposarcoma cells. (a) Western blot analysis of Eag1 protein levels in liposarcoma cells treated with Eag1 shRNA. Densitometric analysis of the bolts with GAPDH as loading control. The results are expressed as mean ± SD (n = 3). (b) The proliferation of liposarcoma cells is determined by CCK-8 assay after treatment with imipramine or Eag1 shRNA. The proliferation of liposarcoma cells is significantly reduced after treatment with imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 6). (c) The tumorigenicity of liposarcoma cells is determined by colony formation assay. The tumorigenicity of liposarcoma cells is significantly reduced after treatment with 20 μM imipramine or Eag1 shRNA. Data are presented as mean ± SD (n = 3). ∗∗P < 0.01.
Mentions: To characterize the oncogenic role of Eag1 in liposarcoma, we first inhibited Eag1 expression by Eag1 shRNA. As shown in Figure 3(a), Eag1 protein expression was effectively suppressed by Ad5-Eag1-shRNA. Next the effect of Eag1 knockdown on cell proliferation was determined by CCK-8 assay. The results revealed that the proliferation of SW-872 and 93T449 cells was inhibited by 39% and 31%, respectively, after Ad5-Eag1-shRNA infection. To confirm the oncogenic role of Eag1, we treated liposarcoma cells with imipramine, a nonspecific blocker of Eag1 activity. The results showed that the proliferation of SW-872 and 93T449 cells was inhibited by 28% and 22%, respectively, after the treatment with 20 μM imipramine (Figure 3(b)). Similar results were obtained from the colony formation experiment (Figure 3(c)). Collectively, these results suggest that Eag1 promotes the proliferation of liposarcoma cells in vitro.

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