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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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Bosutinib pretreatment abolished accumulation of nuclear oxidized PTEN, rescuing tumorigenic phenotypes in shSDHD-transfected FTC133-PTEN wild-type cells, but not in shSDHD-transfected FTC236-PTEN  cells. (A) Nuclear oxidized PTEN was measured by western blot in FTC-133-PTEN wild-type cells, in the presence or absence of H2O2, with or without bosutinib pretreatment. Western blots were performed independently five times. (B) Wound healing migration rates were measured in shSDHD-transfected FTC133-PTEN wild-type cells and shCON-transfected FTC133-PTEN wild-type cells with or without bosutinib pretreatment (upper); wound healing migration rates were measured in shSDHD-transfected FTC236-PTEN  and shCON-tranfected FTC236-PTEN  cells with or without bosutinib pretreatment (lower). The results are the mean ± SE of three independent experiments with n = 5 in each condition. *P < 0.05. (C) TUNEL staining showing apoptotic cells in FTC133-PTEN wild-type cells (upper panels), and in FTC236-PTEN  cells (lower panels) transfected with either shSDHD or shCON, in the presence or absence of H2O2, with or without bosutinib. Magnification: ×200. Experiments were performed four independent times.
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DDU425F2: Bosutinib pretreatment abolished accumulation of nuclear oxidized PTEN, rescuing tumorigenic phenotypes in shSDHD-transfected FTC133-PTEN wild-type cells, but not in shSDHD-transfected FTC236-PTEN cells. (A) Nuclear oxidized PTEN was measured by western blot in FTC-133-PTEN wild-type cells, in the presence or absence of H2O2, with or without bosutinib pretreatment. Western blots were performed independently five times. (B) Wound healing migration rates were measured in shSDHD-transfected FTC133-PTEN wild-type cells and shCON-transfected FTC133-PTEN wild-type cells with or without bosutinib pretreatment (upper); wound healing migration rates were measured in shSDHD-transfected FTC236-PTEN and shCON-tranfected FTC236-PTEN cells with or without bosutinib pretreatment (lower). The results are the mean ± SE of three independent experiments with n = 5 in each condition. *P < 0.05. (C) TUNEL staining showing apoptotic cells in FTC133-PTEN wild-type cells (upper panels), and in FTC236-PTEN cells (lower panels) transfected with either shSDHD or shCON, in the presence or absence of H2O2, with or without bosutinib. Magnification: ×200. Experiments were performed four independent times.

Mentions: To confirm our observations in SDHD-variant-bearing FTC133-PTEN wild-type and FTC236-PTEN cell lines, we silenced SDHD expression in FTC133-PTEN wild-type and FTC236-PTEN cell lines. More oxidized PTEN was observed in the nuclear fraction upon H2O2 treatment in FTC133-PTEN wild-type cells. Bosutinib pretreatment was associated with lack of oxidized PTEN in both SDHD-silenced and control (sham transfected) cells (Fig. 2A). Consistent with our observations in SDHD-G12S/H50R cells, apoptosis induced by H2O2 was noted in SDHD-silenced FTC133-PTEN wild-type cells that were bosutinib pretreated. Apoptosis resistance in SDHD-silenced FTC236-PTEN cells was not affected by bosutinib (Fig. 2B). Induced migration in SDHD-silenced FTC236-PTEN cells was dramatically decreased with bosutinib pretreatment, whereas SDHD-silenced FTC236-PTEN cells showed no significant change in migration even with bosutinib pretreatment (Fig. 2C).Figure 2.


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)

Bosutinib pretreatment abolished accumulation of nuclear oxidized PTEN, rescuing tumorigenic phenotypes in shSDHD-transfected FTC133-PTEN wild-type cells, but not in shSDHD-transfected FTC236-PTEN  cells. (A) Nuclear oxidized PTEN was measured by western blot in FTC-133-PTEN wild-type cells, in the presence or absence of H2O2, with or without bosutinib pretreatment. Western blots were performed independently five times. (B) Wound healing migration rates were measured in shSDHD-transfected FTC133-PTEN wild-type cells and shCON-transfected FTC133-PTEN wild-type cells with or without bosutinib pretreatment (upper); wound healing migration rates were measured in shSDHD-transfected FTC236-PTEN  and shCON-tranfected FTC236-PTEN  cells with or without bosutinib pretreatment (lower). The results are the mean ± SE of three independent experiments with n = 5 in each condition. *P < 0.05. (C) TUNEL staining showing apoptotic cells in FTC133-PTEN wild-type cells (upper panels), and in FTC236-PTEN  cells (lower panels) transfected with either shSDHD or shCON, in the presence or absence of H2O2, with or without bosutinib. Magnification: ×200. Experiments were performed four independent times.
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DDU425F2: Bosutinib pretreatment abolished accumulation of nuclear oxidized PTEN, rescuing tumorigenic phenotypes in shSDHD-transfected FTC133-PTEN wild-type cells, but not in shSDHD-transfected FTC236-PTEN cells. (A) Nuclear oxidized PTEN was measured by western blot in FTC-133-PTEN wild-type cells, in the presence or absence of H2O2, with or without bosutinib pretreatment. Western blots were performed independently five times. (B) Wound healing migration rates were measured in shSDHD-transfected FTC133-PTEN wild-type cells and shCON-transfected FTC133-PTEN wild-type cells with or without bosutinib pretreatment (upper); wound healing migration rates were measured in shSDHD-transfected FTC236-PTEN and shCON-tranfected FTC236-PTEN cells with or without bosutinib pretreatment (lower). The results are the mean ± SE of three independent experiments with n = 5 in each condition. *P < 0.05. (C) TUNEL staining showing apoptotic cells in FTC133-PTEN wild-type cells (upper panels), and in FTC236-PTEN cells (lower panels) transfected with either shSDHD or shCON, in the presence or absence of H2O2, with or without bosutinib. Magnification: ×200. Experiments were performed four independent times.
Mentions: To confirm our observations in SDHD-variant-bearing FTC133-PTEN wild-type and FTC236-PTEN cell lines, we silenced SDHD expression in FTC133-PTEN wild-type and FTC236-PTEN cell lines. More oxidized PTEN was observed in the nuclear fraction upon H2O2 treatment in FTC133-PTEN wild-type cells. Bosutinib pretreatment was associated with lack of oxidized PTEN in both SDHD-silenced and control (sham transfected) cells (Fig. 2A). Consistent with our observations in SDHD-G12S/H50R cells, apoptosis induced by H2O2 was noted in SDHD-silenced FTC133-PTEN wild-type cells that were bosutinib pretreated. Apoptosis resistance in SDHD-silenced FTC236-PTEN cells was not affected by bosutinib (Fig. 2B). Induced migration in SDHD-silenced FTC236-PTEN cells was dramatically decreased with bosutinib pretreatment, whereas SDHD-silenced FTC236-PTEN cells showed no significant change in migration even with bosutinib pretreatment (Fig. 2C).Figure 2.

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