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Se-methylselenocysteine inhibits apoptosis induced by clusterin knockdown in neuroblastoma N2a and SH-SY5Y cell lines.

Wang C, Zeng Z, Liu Q, Zhang R, Ni J - Int J Mol Sci (2014)

Bottom Line: Its biological function is also associated with cell apoptosis.Se-methylselenocysteine (MSC) at an optimum concentration of 1 μM could reverse the alteration in antioxidative capacity, Bcl2/Bax ratio and caspase-8 activity caused by Clu-knockdown, thus inhibiting apoptosis and maintaining cell viability.The results hereby imply the potentiality of Clu and Se in neuroprotection.

View Article: PubMed Central - PubMed

Affiliation: Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China. raulw2003@163.com.

ABSTRACT
Apoptosis, as a programmed cell death process, is essential for the maintenance of tissue function in organisms. Alteration of this process is linked to many diseases. Over-expression of clusterin (Clu) can antagonize apoptosis in various cells. Selenium (Se) is an essential trace element for human health. Its biological function is also associated with cell apoptosis. To explore the function of Clu and the impact of Se in the process of apoptosis, several short-hairpin RNAs (shRNA) were designed for the construction of two sets of recombinant plasmids: one set for plasmid-transfection of mouse neuroblastoma N2a cells (N2a cells); and the other set for lentiviral infection of human neuroblastoma SH-SY5Y cells (SH-SY5Y cells). These shRNAs specifically and efficiently interfered with the intracellular expression of Clu at both the mRNA and protein levels. The Clu-knockdown cells showed apoptosis-related features, including down-regulation of antioxidative capacity and the Bcl-2/Bax ratio and up-regulation of caspase-8 activity. Se-methylselenocysteine (MSC) at an optimum concentration of 1 μM could reverse the alteration in antioxidative capacity, Bcl2/Bax ratio and caspase-8 activity caused by Clu-knockdown, thus inhibiting apoptosis and maintaining cell viability. The results hereby imply the potentiality of Clu and Se in neuroprotection.

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Related in: MedlinePlus

Altered expression levels of apoptosis-related genes Bax, Bcl-2 and Bad in the Clu-knockdown N2a cells treated with 1 μM MSC. Cells transfected with the plasmid pGPU-Clu-mus-sh644 (sh644) were used as the control. (A) Real-time PCR detection of Bax, Bcl-2 and Bad mRNA expression; (B) Western blot analyses of Bax and Bcl-2 protein expression. **p < 0.01 and ***p < 0.001 vs. the control.
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ijms-15-21331-f006: Altered expression levels of apoptosis-related genes Bax, Bcl-2 and Bad in the Clu-knockdown N2a cells treated with 1 μM MSC. Cells transfected with the plasmid pGPU-Clu-mus-sh644 (sh644) were used as the control. (A) Real-time PCR detection of Bax, Bcl-2 and Bad mRNA expression; (B) Western blot analyses of Bax and Bcl-2 protein expression. **p < 0.01 and ***p < 0.001 vs. the control.

Mentions: The effects of MSC on apoptosis-related proteins in the Clu-knockdown cells were also investigated. After transfecting plasmid sh644 for 48 h in N2a cells, MSC (1 μM) was added into the culture medium to treat cells for another 24 h. Then, the cells were collected and extracted for RNAs and proteins for real-time PCR and western blot analyses. At the mRNA level, the expression level of Bcl-2 was increased to 132% (p < 0.01), while the levels of Bax and Bad were decreased to 91% (p < 0.01) and 90% (p < 0.001), respectively, compared to the sh644 plasmid-transfected cells (Figure 6A). Obviously, MSC also increased Bcl-2 expression and decreased Bax expression at protein levels (Figure 6B).


Se-methylselenocysteine inhibits apoptosis induced by clusterin knockdown in neuroblastoma N2a and SH-SY5Y cell lines.

Wang C, Zeng Z, Liu Q, Zhang R, Ni J - Int J Mol Sci (2014)

Altered expression levels of apoptosis-related genes Bax, Bcl-2 and Bad in the Clu-knockdown N2a cells treated with 1 μM MSC. Cells transfected with the plasmid pGPU-Clu-mus-sh644 (sh644) were used as the control. (A) Real-time PCR detection of Bax, Bcl-2 and Bad mRNA expression; (B) Western blot analyses of Bax and Bcl-2 protein expression. **p < 0.01 and ***p < 0.001 vs. the control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4264228&req=5

ijms-15-21331-f006: Altered expression levels of apoptosis-related genes Bax, Bcl-2 and Bad in the Clu-knockdown N2a cells treated with 1 μM MSC. Cells transfected with the plasmid pGPU-Clu-mus-sh644 (sh644) were used as the control. (A) Real-time PCR detection of Bax, Bcl-2 and Bad mRNA expression; (B) Western blot analyses of Bax and Bcl-2 protein expression. **p < 0.01 and ***p < 0.001 vs. the control.
Mentions: The effects of MSC on apoptosis-related proteins in the Clu-knockdown cells were also investigated. After transfecting plasmid sh644 for 48 h in N2a cells, MSC (1 μM) was added into the culture medium to treat cells for another 24 h. Then, the cells were collected and extracted for RNAs and proteins for real-time PCR and western blot analyses. At the mRNA level, the expression level of Bcl-2 was increased to 132% (p < 0.01), while the levels of Bax and Bad were decreased to 91% (p < 0.01) and 90% (p < 0.001), respectively, compared to the sh644 plasmid-transfected cells (Figure 6A). Obviously, MSC also increased Bcl-2 expression and decreased Bax expression at protein levels (Figure 6B).

Bottom Line: Its biological function is also associated with cell apoptosis.Se-methylselenocysteine (MSC) at an optimum concentration of 1 μM could reverse the alteration in antioxidative capacity, Bcl2/Bax ratio and caspase-8 activity caused by Clu-knockdown, thus inhibiting apoptosis and maintaining cell viability.The results hereby imply the potentiality of Clu and Se in neuroprotection.

View Article: PubMed Central - PubMed

Affiliation: Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China. raulw2003@163.com.

ABSTRACT
Apoptosis, as a programmed cell death process, is essential for the maintenance of tissue function in organisms. Alteration of this process is linked to many diseases. Over-expression of clusterin (Clu) can antagonize apoptosis in various cells. Selenium (Se) is an essential trace element for human health. Its biological function is also associated with cell apoptosis. To explore the function of Clu and the impact of Se in the process of apoptosis, several short-hairpin RNAs (shRNA) were designed for the construction of two sets of recombinant plasmids: one set for plasmid-transfection of mouse neuroblastoma N2a cells (N2a cells); and the other set for lentiviral infection of human neuroblastoma SH-SY5Y cells (SH-SY5Y cells). These shRNAs specifically and efficiently interfered with the intracellular expression of Clu at both the mRNA and protein levels. The Clu-knockdown cells showed apoptosis-related features, including down-regulation of antioxidative capacity and the Bcl-2/Bax ratio and up-regulation of caspase-8 activity. Se-methylselenocysteine (MSC) at an optimum concentration of 1 μM could reverse the alteration in antioxidative capacity, Bcl2/Bax ratio and caspase-8 activity caused by Clu-knockdown, thus inhibiting apoptosis and maintaining cell viability. The results hereby imply the potentiality of Clu and Se in neuroprotection.

Show MeSH
Related in: MedlinePlus