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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

Accumulation of oxidized nuclear PTEN from SDHD-G12S and SDHD-H50R was abolished with bosutinib pretreatment in FTC133-PTEN wild-type cell, but not in FTC236-PTEN  cells. (A) Lipid peroxidation levels were measured in FTC133-PTEN wild-type and FTC23-PTEN  cells transfected with SDHD-wild type, -G12S and -H50R. Means ± SE from three representative experiments with n = 3 in each condition. (B) Nuclear PTEN was measured in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R, respectively, after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed three independent times, and the best blot was shown. (C) Nuclear oxidized PTEN was measured in FTC133-PTEN wild-type cells after transfection with wild type, G12S and H50R SDHD constructs after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed four independent times and the best blot was shown. (D) Apoptotic rates in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment (left). Apoptotic rates in FTC236-PTEN  cells with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment. Results are the mean ± SE of two independent experiments with n = 3 under each condition. (E) Wound healing migration rates were measured in FTC133-PTEN wild type containing SDHD-WT, -G12S or -H50R with/without bosutinib pretreatment (left) and in FTC236-PTEN  transfected with SDHD-wild type, -G12S and -H50R with or without bosutinib pretreatment (right). The results are the mean ± SE of three independent experiments with n = 5 in each condition. **P < 0.005, *P < 0.05.
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DDU425F1: Accumulation of oxidized nuclear PTEN from SDHD-G12S and SDHD-H50R was abolished with bosutinib pretreatment in FTC133-PTEN wild-type cell, but not in FTC236-PTEN cells. (A) Lipid peroxidation levels were measured in FTC133-PTEN wild-type and FTC23-PTEN cells transfected with SDHD-wild type, -G12S and -H50R. Means ± SE from three representative experiments with n = 3 in each condition. (B) Nuclear PTEN was measured in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R, respectively, after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed three independent times, and the best blot was shown. (C) Nuclear oxidized PTEN was measured in FTC133-PTEN wild-type cells after transfection with wild type, G12S and H50R SDHD constructs after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed four independent times and the best blot was shown. (D) Apoptotic rates in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment (left). Apoptotic rates in FTC236-PTEN cells with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment. Results are the mean ± SE of two independent experiments with n = 3 under each condition. (E) Wound healing migration rates were measured in FTC133-PTEN wild type containing SDHD-WT, -G12S or -H50R with/without bosutinib pretreatment (left) and in FTC236-PTEN transfected with SDHD-wild type, -G12S and -H50R with or without bosutinib pretreatment (right). The results are the mean ± SE of three independent experiments with n = 5 in each condition. **P < 0.005, *P < 0.05.

Mentions: We previously found that cellular ROS is significantly increased in CS/CSL patient samples harboring germline SDHx variants compared with normal controls (16). To determine if the SDHD variants in thyroid cancer cells can result in damage to lipids by, e.g. lipid peroxidation, we measured the byproducts of polyunsaturated fatty acid peroxides upon decomposition, namely, malondialdehyde (MDA) and 4-hydroxyalkenals (24) in two thyroid cancer cell lines follicular thyroid carcinoma (FTC) 133-PTEN wild-type and FTC236-PTEN cells transfected with SDHD-G12S or -H50R. Compared with control SDHD-wild-type (25) transfected cells, no significant increase of lipid peroxidation was observed in FTC133-PTEN wild-type cells with SDHD-G12S or SDHD-H50R. In contrast, a slight increase in lipid peroxidation was observed in SDHD-G12S or SDHD-H50R transfected FTC236-PTEN cells (Fig. 1A). We therefore surmised that wild-type PTEN in FTC133-PTEN wild type could play a protective role against SDHD variant-induced oxidative stress.Figure 1.


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)

Accumulation of oxidized nuclear PTEN from SDHD-G12S and SDHD-H50R was abolished with bosutinib pretreatment in FTC133-PTEN wild-type cell, but not in FTC236-PTEN  cells. (A) Lipid peroxidation levels were measured in FTC133-PTEN wild-type and FTC23-PTEN  cells transfected with SDHD-wild type, -G12S and -H50R. Means ± SE from three representative experiments with n = 3 in each condition. (B) Nuclear PTEN was measured in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R, respectively, after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed three independent times, and the best blot was shown. (C) Nuclear oxidized PTEN was measured in FTC133-PTEN wild-type cells after transfection with wild type, G12S and H50R SDHD constructs after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed four independent times and the best blot was shown. (D) Apoptotic rates in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment (left). Apoptotic rates in FTC236-PTEN  cells with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment. Results are the mean ± SE of two independent experiments with n = 3 under each condition. (E) Wound healing migration rates were measured in FTC133-PTEN wild type containing SDHD-WT, -G12S or -H50R with/without bosutinib pretreatment (left) and in FTC236-PTEN  transfected with SDHD-wild type, -G12S and -H50R with or without bosutinib pretreatment (right). The results are the mean ± SE of three independent experiments with n = 5 in each condition. **P < 0.005, *P < 0.05.
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DDU425F1: Accumulation of oxidized nuclear PTEN from SDHD-G12S and SDHD-H50R was abolished with bosutinib pretreatment in FTC133-PTEN wild-type cell, but not in FTC236-PTEN cells. (A) Lipid peroxidation levels were measured in FTC133-PTEN wild-type and FTC23-PTEN cells transfected with SDHD-wild type, -G12S and -H50R. Means ± SE from three representative experiments with n = 3 in each condition. (B) Nuclear PTEN was measured in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R, respectively, after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed three independent times, and the best blot was shown. (C) Nuclear oxidized PTEN was measured in FTC133-PTEN wild-type cells after transfection with wild type, G12S and H50R SDHD constructs after H2O2 exposure with or without bosutinib pretreatment. Western blots were performed four independent times and the best blot was shown. (D) Apoptotic rates in FTC133-PTEN wild-type cells transfected with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment (left). Apoptotic rates in FTC236-PTEN cells with SDHD-wild type, -G12S and -H50R with or without H2O2, with or without bosutinib pretreatment. Results are the mean ± SE of two independent experiments with n = 3 under each condition. (E) Wound healing migration rates were measured in FTC133-PTEN wild type containing SDHD-WT, -G12S or -H50R with/without bosutinib pretreatment (left) and in FTC236-PTEN transfected with SDHD-wild type, -G12S and -H50R with or without bosutinib pretreatment (right). The results are the mean ± SE of three independent experiments with n = 5 in each condition. **P < 0.005, *P < 0.05.
Mentions: We previously found that cellular ROS is significantly increased in CS/CSL patient samples harboring germline SDHx variants compared with normal controls (16). To determine if the SDHD variants in thyroid cancer cells can result in damage to lipids by, e.g. lipid peroxidation, we measured the byproducts of polyunsaturated fatty acid peroxides upon decomposition, namely, malondialdehyde (MDA) and 4-hydroxyalkenals (24) in two thyroid cancer cell lines follicular thyroid carcinoma (FTC) 133-PTEN wild-type and FTC236-PTEN cells transfected with SDHD-G12S or -H50R. Compared with control SDHD-wild-type (25) transfected cells, no significant increase of lipid peroxidation was observed in FTC133-PTEN wild-type cells with SDHD-G12S or SDHD-H50R. In contrast, a slight increase in lipid peroxidation was observed in SDHD-G12S or SDHD-H50R transfected FTC236-PTEN cells (Fig. 1A). We therefore surmised that wild-type PTEN in FTC133-PTEN wild type could play a protective role against SDHD variant-induced oxidative stress.Figure 1.

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