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Cowden syndrome-associated germline SDHD variants alter PTEN nuclear translocation through SRC-induced PTEN oxidation.

Yu W, He X, Ni Y, Ngeow J, Eng C - Hum. Mol. Genet. (2014)

Bottom Line: However, very little is known about the underlying crosstalk between SDHD and PTEN in CS-associated thyroid cancer.We show that SRC inhibition could rescue SDHD dysfunction-induced cellular phenotype and tumorigenesis only when wild-type PTEN is expressed, in thyroid cancer lines.Patient lymphoblast cells carrying either SDHD-G12S or SDHD-H50R also show increased nuclear PTEN and more oxidized PTEN after hydrogen peroxide treatment.

View Article: PubMed Central - PubMed

Affiliation: Genomic Medicine Institute, Learner Research Institute.

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Related in: MedlinePlus

HIF-1α was stabilized with H2O2 treatment and inhibited by SRC inhibitor bosutinib in FTC133-PTEN wild-type cells. (A) Western blot of HIF-1α expression in SDHD-wild type, -G12S or -H50R transfected into FTC133-PTEN wild-type cells, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (B) HIF-1α expression in FTC133-PTEN wild-type cells after shSDHD transfection or shCON control-transfection with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (C) HIF-1α expression in FTC133-PTEN wild-type cells transfected with shSRC or shCON control construct, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of two independent experiments. (D) Wound healing migration rates with or without HIF-1 inhibitor chetomin pretreatment in FTC133-PTEN wild-type cells (left) and in FTC236-PTEN  cells (right). The results are the mean ± SE of two independent experiments with n = 4 in each condition. (E) Wound healing migration rates in wild-type PTEN transfected FTC236-PTEN  cells with or without bosutinib. The results are the mean ± SE of three independent experiments with n = 3 in each condition. (F) Wound healing migration rates in PTEN knockout FTC133-PTEN wild-type cells with or without bosutinib. The results are the mean ± SE of two repeated experiments with n = 3 in each condition. *P < 0.05.
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DDU425F4: HIF-1α was stabilized with H2O2 treatment and inhibited by SRC inhibitor bosutinib in FTC133-PTEN wild-type cells. (A) Western blot of HIF-1α expression in SDHD-wild type, -G12S or -H50R transfected into FTC133-PTEN wild-type cells, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (B) HIF-1α expression in FTC133-PTEN wild-type cells after shSDHD transfection or shCON control-transfection with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (C) HIF-1α expression in FTC133-PTEN wild-type cells transfected with shSRC or shCON control construct, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of two independent experiments. (D) Wound healing migration rates with or without HIF-1 inhibitor chetomin pretreatment in FTC133-PTEN wild-type cells (left) and in FTC236-PTEN cells (right). The results are the mean ± SE of two independent experiments with n = 4 in each condition. (E) Wound healing migration rates in wild-type PTEN transfected FTC236-PTEN cells with or without bosutinib. The results are the mean ± SE of three independent experiments with n = 3 in each condition. (F) Wound healing migration rates in PTEN knockout FTC133-PTEN wild-type cells with or without bosutinib. The results are the mean ± SE of two repeated experiments with n = 3 in each condition. *P < 0.05.

Mentions: To further identify the effect of our SRC kinase inhibitor on thyroid cancer cell lines, we examined the activation of HIF-1α by ROS with or without SRC inhibition. In FTC133-PTEN wild-type cells expressing SDHD-G12S or SDHD-H50R, H2O2 treatment dramatically induced HIF-1α expression, with the levels of increased expression in SDHD-G12S or SDHD-H50R-containing cells much more dramatic compared with those from control (SDHD-WT) cells. Pretreatment with bosutinib was associated with blockade of the increased HIF-1α expression from H2O2 exposure in both SDHD-G12S/H50R expressing cells and control (SDHD-wild-type) cells (Fig. 4A). Consistently, we observed further increases of HIF-1α expression in SDHD-silenced cells compared with control cells (transfected with empty plasmid), and this increased HIF-1α expression was also blocked by bosutinib pretreatment (Fig. 4B).Figure 4.


Cowden syndrome-associated germline SDHD variants alter PTEN nuclear translocation through SRC-induced PTEN oxidation.

Yu W, He X, Ni Y, Ngeow J, Eng C - Hum. Mol. Genet. (2014)

HIF-1α was stabilized with H2O2 treatment and inhibited by SRC inhibitor bosutinib in FTC133-PTEN wild-type cells. (A) Western blot of HIF-1α expression in SDHD-wild type, -G12S or -H50R transfected into FTC133-PTEN wild-type cells, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (B) HIF-1α expression in FTC133-PTEN wild-type cells after shSDHD transfection or shCON control-transfection with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (C) HIF-1α expression in FTC133-PTEN wild-type cells transfected with shSRC or shCON control construct, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of two independent experiments. (D) Wound healing migration rates with or without HIF-1 inhibitor chetomin pretreatment in FTC133-PTEN wild-type cells (left) and in FTC236-PTEN  cells (right). The results are the mean ± SE of two independent experiments with n = 4 in each condition. (E) Wound healing migration rates in wild-type PTEN transfected FTC236-PTEN  cells with or without bosutinib. The results are the mean ± SE of three independent experiments with n = 3 in each condition. (F) Wound healing migration rates in PTEN knockout FTC133-PTEN wild-type cells with or without bosutinib. The results are the mean ± SE of two repeated experiments with n = 3 in each condition. *P < 0.05.
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DDU425F4: HIF-1α was stabilized with H2O2 treatment and inhibited by SRC inhibitor bosutinib in FTC133-PTEN wild-type cells. (A) Western blot of HIF-1α expression in SDHD-wild type, -G12S or -H50R transfected into FTC133-PTEN wild-type cells, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (B) HIF-1α expression in FTC133-PTEN wild-type cells after shSDHD transfection or shCON control-transfection with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of three independent experiments. (C) HIF-1α expression in FTC133-PTEN wild-type cells transfected with shSRC or shCON control construct, with or without H2O2, with or without bosutinib. The bar graph summarizes the relative expression of two independent experiments. (D) Wound healing migration rates with or without HIF-1 inhibitor chetomin pretreatment in FTC133-PTEN wild-type cells (left) and in FTC236-PTEN cells (right). The results are the mean ± SE of two independent experiments with n = 4 in each condition. (E) Wound healing migration rates in wild-type PTEN transfected FTC236-PTEN cells with or without bosutinib. The results are the mean ± SE of three independent experiments with n = 3 in each condition. (F) Wound healing migration rates in PTEN knockout FTC133-PTEN wild-type cells with or without bosutinib. The results are the mean ± SE of two repeated experiments with n = 3 in each condition. *P < 0.05.
Mentions: To further identify the effect of our SRC kinase inhibitor on thyroid cancer cell lines, we examined the activation of HIF-1α by ROS with or without SRC inhibition. In FTC133-PTEN wild-type cells expressing SDHD-G12S or SDHD-H50R, H2O2 treatment dramatically induced HIF-1α expression, with the levels of increased expression in SDHD-G12S or SDHD-H50R-containing cells much more dramatic compared with those from control (SDHD-WT) cells. Pretreatment with bosutinib was associated with blockade of the increased HIF-1α expression from H2O2 exposure in both SDHD-G12S/H50R expressing cells and control (SDHD-wild-type) cells (Fig. 4A). Consistently, we observed further increases of HIF-1α expression in SDHD-silenced cells compared with control cells (transfected with empty plasmid), and this increased HIF-1α expression was also blocked by bosutinib pretreatment (Fig. 4B).Figure 4.

Bottom Line: However, very little is known about the underlying crosstalk between SDHD and PTEN in CS-associated thyroid cancer.We show that SRC inhibition could rescue SDHD dysfunction-induced cellular phenotype and tumorigenesis only when wild-type PTEN is expressed, in thyroid cancer lines.Patient lymphoblast cells carrying either SDHD-G12S or SDHD-H50R also show increased nuclear PTEN and more oxidized PTEN after hydrogen peroxide treatment.

View Article: PubMed Central - PubMed

Affiliation: Genomic Medicine Institute, Learner Research Institute.

Show MeSH
Related in: MedlinePlus