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Relevance and therapeutic possibility of PTEN-long in renal cell carcinoma.

Wang H, Zhang P, Lin C, Yu Q, Wu J, Wang L, Cui Y, Wang K, Gao Z, Li H - PLoS ONE (2015)

Bottom Line: We found that the protein levels of PTEN-Long were drastically reduced in ccRCC, which was correlated with increased levels of phosphorylated Akt (pAkt).When purified PTEN-Long was added into cultured 786-0 cells, it entered cells, blocked Akt activation, and induced apoptosis involving Caspase 3 cleavage.Furthermore, PTEN-Long inhibited proliferation of 786-0 cells in xenograft mouse model.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College, Institute of Urology, Zhifu, Yantai, Shandong, 264000, People's Republic of China.

ABSTRACT
PTEN-Long is a translational variant of PTEN (Phosphatase and Tensin Homolog). Like PTEN, PTEN-Long is able to antagonize the PI3K-Akt pathway and inhibits tumor growth. In this study, we investigated the role PTEN-Long plays in the development and progression of clear cell renal cell carcinoma (ccRCC) and explored the therapeutic possibility using proteinaceous PTEN-Long to treat ccRCC. We found that the protein levels of PTEN-Long were drastically reduced in ccRCC, which was correlated with increased levels of phosphorylated Akt (pAkt). Gain of function experiments showed overexpression of PTEN-Long in the ccRCC cell line 786-0 suppressed PI3K-Akt signaling, inhibited cell proliferation, migration and invasion, and eventually induced cell death. When purified PTEN-Long was added into cultured 786-0 cells, it entered cells, blocked Akt activation, and induced apoptosis involving Caspase 3 cleavage. Furthermore, PTEN-Long inhibited proliferation of 786-0 cells in xenograft mouse model. Our results implicated that understanding the roles of PTEN-Long in renal cell carcinogenesis has therapeutic significance.

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Related in: MedlinePlus

Overexpression of PTEN-Long inhibits cell migration and reduces cell invasion.A, 786-0 cells stably expressing indicated proteins were subjected to in vitro wounding and followed by phase-contrast imaging. The fields in between the white lines indicate the wound track. The scale bar represents 20 µm. B, The number of cells migrated in the wound track from 6 random separate fields were counted for each cell and expressed as a mean ± SD (error bars). C, Invasive properties of 786-0 cells were inhibited by 30% by PTEN overexpression and by 25% by PTEN-Long overexpression, as compared to empty vector control cells.
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pone-0114250-g004: Overexpression of PTEN-Long inhibits cell migration and reduces cell invasion.A, 786-0 cells stably expressing indicated proteins were subjected to in vitro wounding and followed by phase-contrast imaging. The fields in between the white lines indicate the wound track. The scale bar represents 20 µm. B, The number of cells migrated in the wound track from 6 random separate fields were counted for each cell and expressed as a mean ± SD (error bars). C, Invasive properties of 786-0 cells were inhibited by 30% by PTEN overexpression and by 25% by PTEN-Long overexpression, as compared to empty vector control cells.

Mentions: Next, we investigated the contribution of PTEN-Long to the migratory properties of 786-0 by scratch assay. First, we showed that the expression of PTEN inhibited the ability of 786-0 cells to migrate into a wound track, as compared with vector control cells (Fig. 4A, rows 1 and 2). Expression of PTEN-Long also inhibited migration of cells (Fig. 4A, rows 4) and this function was dependent on its lipid phosphatase activity, as both PTENG129R and PTEN-LongG302R mutants failed to produce the same results (Fig. 4A, row 3 and 5). Quantitative analysis of cells migrated into wound area 11 h after scratching revealed a roughly 35% (P<0.001) inhibition in migration of 786-0 cells into the wound track by overexpression of PTEN-Long or PTEN (Fig. 4B). Because the G to R mutants of PTEN and PTEN-Long are still theoretically functional as protein phosphatase, failure of the G to R mutants to inhibit migration of 786-0 indicates that protein phosphatase activity of PTEN and PTEN-Long is not sufficient for this function.


Relevance and therapeutic possibility of PTEN-long in renal cell carcinoma.

Wang H, Zhang P, Lin C, Yu Q, Wu J, Wang L, Cui Y, Wang K, Gao Z, Li H - PLoS ONE (2015)

Overexpression of PTEN-Long inhibits cell migration and reduces cell invasion.A, 786-0 cells stably expressing indicated proteins were subjected to in vitro wounding and followed by phase-contrast imaging. The fields in between the white lines indicate the wound track. The scale bar represents 20 µm. B, The number of cells migrated in the wound track from 6 random separate fields were counted for each cell and expressed as a mean ± SD (error bars). C, Invasive properties of 786-0 cells were inhibited by 30% by PTEN overexpression and by 25% by PTEN-Long overexpression, as compared to empty vector control cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340966&req=5

pone-0114250-g004: Overexpression of PTEN-Long inhibits cell migration and reduces cell invasion.A, 786-0 cells stably expressing indicated proteins were subjected to in vitro wounding and followed by phase-contrast imaging. The fields in between the white lines indicate the wound track. The scale bar represents 20 µm. B, The number of cells migrated in the wound track from 6 random separate fields were counted for each cell and expressed as a mean ± SD (error bars). C, Invasive properties of 786-0 cells were inhibited by 30% by PTEN overexpression and by 25% by PTEN-Long overexpression, as compared to empty vector control cells.
Mentions: Next, we investigated the contribution of PTEN-Long to the migratory properties of 786-0 by scratch assay. First, we showed that the expression of PTEN inhibited the ability of 786-0 cells to migrate into a wound track, as compared with vector control cells (Fig. 4A, rows 1 and 2). Expression of PTEN-Long also inhibited migration of cells (Fig. 4A, rows 4) and this function was dependent on its lipid phosphatase activity, as both PTENG129R and PTEN-LongG302R mutants failed to produce the same results (Fig. 4A, row 3 and 5). Quantitative analysis of cells migrated into wound area 11 h after scratching revealed a roughly 35% (P<0.001) inhibition in migration of 786-0 cells into the wound track by overexpression of PTEN-Long or PTEN (Fig. 4B). Because the G to R mutants of PTEN and PTEN-Long are still theoretically functional as protein phosphatase, failure of the G to R mutants to inhibit migration of 786-0 indicates that protein phosphatase activity of PTEN and PTEN-Long is not sufficient for this function.

Bottom Line: We found that the protein levels of PTEN-Long were drastically reduced in ccRCC, which was correlated with increased levels of phosphorylated Akt (pAkt).When purified PTEN-Long was added into cultured 786-0 cells, it entered cells, blocked Akt activation, and induced apoptosis involving Caspase 3 cleavage.Furthermore, PTEN-Long inhibited proliferation of 786-0 cells in xenograft mouse model.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, the Affiliated Yantai Yuhuangding Hospital of Qingdao University Medical College, Institute of Urology, Zhifu, Yantai, Shandong, 264000, People's Republic of China.

ABSTRACT
PTEN-Long is a translational variant of PTEN (Phosphatase and Tensin Homolog). Like PTEN, PTEN-Long is able to antagonize the PI3K-Akt pathway and inhibits tumor growth. In this study, we investigated the role PTEN-Long plays in the development and progression of clear cell renal cell carcinoma (ccRCC) and explored the therapeutic possibility using proteinaceous PTEN-Long to treat ccRCC. We found that the protein levels of PTEN-Long were drastically reduced in ccRCC, which was correlated with increased levels of phosphorylated Akt (pAkt). Gain of function experiments showed overexpression of PTEN-Long in the ccRCC cell line 786-0 suppressed PI3K-Akt signaling, inhibited cell proliferation, migration and invasion, and eventually induced cell death. When purified PTEN-Long was added into cultured 786-0 cells, it entered cells, blocked Akt activation, and induced apoptosis involving Caspase 3 cleavage. Furthermore, PTEN-Long inhibited proliferation of 786-0 cells in xenograft mouse model. Our results implicated that understanding the roles of PTEN-Long in renal cell carcinogenesis has therapeutic significance.

Show MeSH
Related in: MedlinePlus