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The expression and activity of thioredoxin reductase 1 splice variants v1 and v2 regulate the expression of genes associated with differentiation and adhesion.

Nalvarte I, Damdimopoulos AE, Rüegg J, Spyrou G - Biosci. Rep. (2015)

Bottom Line: The mammalian redox-active selenoprotein thioredoxin reductase (TrxR1) is a main player in redox homoeostasis.Overexpression of these two splice forms resulted in distinctive effects on various aspects of cellular functions including gene regulation patterns, alteration of growth rate, migration and morphology and susceptibility to selenium-induced toxicity.These data suggest that both TXNRD1_v1 and TXNRD1_v2 have distinct roles in differentiation, possibly by altering the expression of the genes associated with differentiation, and further emphasize the importance in distinguishing each unique action of different TrxR1 splice forms, especially when studying the gene silencing or knockout of TrxR1.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences and Nutrition, Karolinska Institute, SE-141 83 Huddinge, Sweden ivan.nalvarte@ki.se.

No MeSH data available.


Related in: MedlinePlus

Comparison between TrxR, fibronectin 1 and cadherin 11 protein levels in SH-SY5Y cells upon selenium and/or ATRA treatment(A) Western blot analysis of TXNRD1_v1, TXNRD1_v2, fibronectin 1 and cadherin 11 in SH-SY5Y cell extracts upon indicated days of 0.2 μM Se, (B) 5 μM ATRA or (C) 0.2 μM Se and 5 μM ATRA treatment. (D, E, F) Left panels show quantified relative protein levels of TXNRD1_v1 and TXNRD1_v2 normalized to Hsp-90 levels for 5 days of respective treatment. Right panels show quantified relative protein levels of fibronectin 1 and Cadherin 11 normalized to Hsp-90 levels with the same treatment. Error bars correspond to S.D., *P<0.05, **P<0.01.
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Figure 6: Comparison between TrxR, fibronectin 1 and cadherin 11 protein levels in SH-SY5Y cells upon selenium and/or ATRA treatment(A) Western blot analysis of TXNRD1_v1, TXNRD1_v2, fibronectin 1 and cadherin 11 in SH-SY5Y cell extracts upon indicated days of 0.2 μM Se, (B) 5 μM ATRA or (C) 0.2 μM Se and 5 μM ATRA treatment. (D, E, F) Left panels show quantified relative protein levels of TXNRD1_v1 and TXNRD1_v2 normalized to Hsp-90 levels for 5 days of respective treatment. Right panels show quantified relative protein levels of fibronectin 1 and Cadherin 11 normalized to Hsp-90 levels with the same treatment. Error bars correspond to S.D., *P<0.05, **P<0.01.

Mentions: When analysing protein levels of TXNRD1_v1 and TXNRD1_v2, we observed a clear rapid increase upon selenium treatment (Figures 6A and 6D). We also observed an increase in cadherin 11 protein levels, but decreased fibronectin 1 protein levels (in contrast with mRNA levels) at days 1–3 of selenium treatment. This could possibly link increased TrxR1 activity with decreased fibronectin 1 levels and thus the decreased migration observed in Figure 3B. At day 8 both fibronectin 1 and cadherin 11 levels were increased, probably due to cell confluency. As expected, ATRA treatment increased both cadherin 11 and fibronectin 1 levels (days 3–5) (Figures 6B and 6E). Also, TXNRD1_v1 and TXNRD1_v2 levels were drastically increased upon ATRA treatment alone (Figures 6B and 6E). However, TXNRD1_v2 returned to control levels at day 2 whereas the levels of TXNRD1_v1 remained high. Interestingly, the increase in TXNRD1_v1 and TXNRD1_v2 levels upon ATRA treatment appeared earlier than the increase in fibronectin 1 and cadherin 11. Treating selenium-pretreated SH-SY5Y cells with selenium and ATRA for 5 days (Figures 6C and 6F) had initially no drastic effects on fibronectin 1 and cadherin 11 levels. The decrease in fibronectin 1 levels after the first selenium treatment was sustained. However, at later time points both fibronectin 1 and cadherin 11 levels were increased (Figures 6C and 6F). Combined selenium and ATRA treatment also increased TXNRD1_v1 and v2 levels, although we could not observe any additive effects. The opposite effect of selenium on fibronectin 1 and cadherin 11 protein levels suggests that TrxR1 activity to be important in keeping cell–cell contacts but not cell-matrix contacts during differentiation. The fast changes in TXNRD1_v2 protein levels compared with the more subtle changes in TXNRD1_v1 protein levels could imply different roles of TXNRD1_v1 and v2 on the cellular response to selenium and ATRA, the expression of fibronectin 1 and cadherin 11, and thus to migration and differentiation.


The expression and activity of thioredoxin reductase 1 splice variants v1 and v2 regulate the expression of genes associated with differentiation and adhesion.

Nalvarte I, Damdimopoulos AE, Rüegg J, Spyrou G - Biosci. Rep. (2015)

Comparison between TrxR, fibronectin 1 and cadherin 11 protein levels in SH-SY5Y cells upon selenium and/or ATRA treatment(A) Western blot analysis of TXNRD1_v1, TXNRD1_v2, fibronectin 1 and cadherin 11 in SH-SY5Y cell extracts upon indicated days of 0.2 μM Se, (B) 5 μM ATRA or (C) 0.2 μM Se and 5 μM ATRA treatment. (D, E, F) Left panels show quantified relative protein levels of TXNRD1_v1 and TXNRD1_v2 normalized to Hsp-90 levels for 5 days of respective treatment. Right panels show quantified relative protein levels of fibronectin 1 and Cadherin 11 normalized to Hsp-90 levels with the same treatment. Error bars correspond to S.D., *P<0.05, **P<0.01.
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Figure 6: Comparison between TrxR, fibronectin 1 and cadherin 11 protein levels in SH-SY5Y cells upon selenium and/or ATRA treatment(A) Western blot analysis of TXNRD1_v1, TXNRD1_v2, fibronectin 1 and cadherin 11 in SH-SY5Y cell extracts upon indicated days of 0.2 μM Se, (B) 5 μM ATRA or (C) 0.2 μM Se and 5 μM ATRA treatment. (D, E, F) Left panels show quantified relative protein levels of TXNRD1_v1 and TXNRD1_v2 normalized to Hsp-90 levels for 5 days of respective treatment. Right panels show quantified relative protein levels of fibronectin 1 and Cadherin 11 normalized to Hsp-90 levels with the same treatment. Error bars correspond to S.D., *P<0.05, **P<0.01.
Mentions: When analysing protein levels of TXNRD1_v1 and TXNRD1_v2, we observed a clear rapid increase upon selenium treatment (Figures 6A and 6D). We also observed an increase in cadherin 11 protein levels, but decreased fibronectin 1 protein levels (in contrast with mRNA levels) at days 1–3 of selenium treatment. This could possibly link increased TrxR1 activity with decreased fibronectin 1 levels and thus the decreased migration observed in Figure 3B. At day 8 both fibronectin 1 and cadherin 11 levels were increased, probably due to cell confluency. As expected, ATRA treatment increased both cadherin 11 and fibronectin 1 levels (days 3–5) (Figures 6B and 6E). Also, TXNRD1_v1 and TXNRD1_v2 levels were drastically increased upon ATRA treatment alone (Figures 6B and 6E). However, TXNRD1_v2 returned to control levels at day 2 whereas the levels of TXNRD1_v1 remained high. Interestingly, the increase in TXNRD1_v1 and TXNRD1_v2 levels upon ATRA treatment appeared earlier than the increase in fibronectin 1 and cadherin 11. Treating selenium-pretreated SH-SY5Y cells with selenium and ATRA for 5 days (Figures 6C and 6F) had initially no drastic effects on fibronectin 1 and cadherin 11 levels. The decrease in fibronectin 1 levels after the first selenium treatment was sustained. However, at later time points both fibronectin 1 and cadherin 11 levels were increased (Figures 6C and 6F). Combined selenium and ATRA treatment also increased TXNRD1_v1 and v2 levels, although we could not observe any additive effects. The opposite effect of selenium on fibronectin 1 and cadherin 11 protein levels suggests that TrxR1 activity to be important in keeping cell–cell contacts but not cell-matrix contacts during differentiation. The fast changes in TXNRD1_v2 protein levels compared with the more subtle changes in TXNRD1_v1 protein levels could imply different roles of TXNRD1_v1 and v2 on the cellular response to selenium and ATRA, the expression of fibronectin 1 and cadherin 11, and thus to migration and differentiation.

Bottom Line: The mammalian redox-active selenoprotein thioredoxin reductase (TrxR1) is a main player in redox homoeostasis.Overexpression of these two splice forms resulted in distinctive effects on various aspects of cellular functions including gene regulation patterns, alteration of growth rate, migration and morphology and susceptibility to selenium-induced toxicity.These data suggest that both TXNRD1_v1 and TXNRD1_v2 have distinct roles in differentiation, possibly by altering the expression of the genes associated with differentiation, and further emphasize the importance in distinguishing each unique action of different TrxR1 splice forms, especially when studying the gene silencing or knockout of TrxR1.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences and Nutrition, Karolinska Institute, SE-141 83 Huddinge, Sweden ivan.nalvarte@ki.se.

No MeSH data available.


Related in: MedlinePlus