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MicroRNA‑34a induces apoptosis in PC12 cells by reducing B‑cell lymphoma 2 and sirtuin‑1 expression.

Lin Q, Mao Y, Song Y, Huang D - Mol Med Rep (2015)

Bottom Line: The effect of miR‑34a in PC12 cells has not yet been reported.It was demonstrated that induction of miR‑34a promotes apoptosis and senescence, inhibits proliferation, and leads to marked alterations in SIRT1, Bcl‑12 and acetyl (ac)‑p53 expression.These data indicate that miR‑34a may be important in neuropathy.

View Article: PubMed Central - PubMed

Affiliation: Department of Rehabilitation Medicine, The First Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510080, P.R. China.

ABSTRACT
MicroRNA‑34a (miR‑34a) is a direct target of p53 and was reported to induce cell cycle arrest, apoptosis and senescence. Inhibition of the NAD‑dependent deacetylase sirtuin‑1 (SIRT1) by miR‑34a leads to an increase in acetylated p53, which promotes cell apoptosis. B‑cell lymphoma 2 (Bcl‑2) is also involved in apoptosis, and was originally characterized with respect to its role in controlling outer mitochondrial membrane integrity. The effect of miR‑34a in PC12 cells has not yet been reported. In the present study, it was hypothesized that Bcl‑2 and SIRT1 may be critical downstream targets of miR‑34a that participate in apoptosis induction. miR‑34a mimics and inhibitors were transfected into PC12 cells, and the apoptosis and proliferation rates were compared between groups. It was demonstrated that induction of miR‑34a promotes apoptosis and senescence, inhibits proliferation, and leads to marked alterations in SIRT1, Bcl‑12 and acetyl (ac)‑p53 expression. These data indicate that miR‑34a may be important in neuropathy.

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Related in: MedlinePlus

Effects of miR-34a mimics and inhibitors on cell apoptosis rates analyzed by fluorescence-activated cell sorting. Cells were dually stained with Annexin V/PI and analyzed by flow cytometry to determine the population of cells in early and late apoptosis stages in the (A) negative control, (B) cells treated with miR-34a inhibitor and (C) cells treated with miR-34a mimics. Left lower quadrant, viable cells; right lower quadrant, early apoptotic cells; right upper quadrant, late apoptotic cells; left upper quadrant, necrotic cells. (D) Quantification of the total percentage of apoptotic cells in each group. **P<0.01, n=3 for each group. miR, microRNA; PI, propidium iodide.
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f2-mmr-12-04-5709: Effects of miR-34a mimics and inhibitors on cell apoptosis rates analyzed by fluorescence-activated cell sorting. Cells were dually stained with Annexin V/PI and analyzed by flow cytometry to determine the population of cells in early and late apoptosis stages in the (A) negative control, (B) cells treated with miR-34a inhibitor and (C) cells treated with miR-34a mimics. Left lower quadrant, viable cells; right lower quadrant, early apoptotic cells; right upper quadrant, late apoptotic cells; left upper quadrant, necrotic cells. (D) Quantification of the total percentage of apoptotic cells in each group. **P<0.01, n=3 for each group. miR, microRNA; PI, propidium iodide.

Mentions: It is well known that miR-34a is an important component of the p53 tumor suppressor protein transcriptional network, which regulates cell proliferation and cell cycle progression. In order to investigate the biological effects of miR-34a in nerve cells, PC12 cells were transiently transfected with miR-34a mimics or miR-34a inhibitors, and the proportions of apoptotic cells were quantified using an Annexin V-FITC/PI dual staining assay. As shown in Fig. 2, after transfection, the total proportion of apoptotic cells in the miR-34a mimic group was significantly increased compared with that of the control group (P<0.01). The apoptotic rate of miR-34a inhibitor group was marginally lower than that of the control group, however, no significant difference was identified (P>0.05). The total apoptotic rates for control, miR-34a inhibitor and miR-34a mimic transfection groups were 12.2±1.06, 11.4±0.34 and 19.2±0.84%, respectively.


MicroRNA‑34a induces apoptosis in PC12 cells by reducing B‑cell lymphoma 2 and sirtuin‑1 expression.

Lin Q, Mao Y, Song Y, Huang D - Mol Med Rep (2015)

Effects of miR-34a mimics and inhibitors on cell apoptosis rates analyzed by fluorescence-activated cell sorting. Cells were dually stained with Annexin V/PI and analyzed by flow cytometry to determine the population of cells in early and late apoptosis stages in the (A) negative control, (B) cells treated with miR-34a inhibitor and (C) cells treated with miR-34a mimics. Left lower quadrant, viable cells; right lower quadrant, early apoptotic cells; right upper quadrant, late apoptotic cells; left upper quadrant, necrotic cells. (D) Quantification of the total percentage of apoptotic cells in each group. **P<0.01, n=3 for each group. miR, microRNA; PI, propidium iodide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581806&req=5

f2-mmr-12-04-5709: Effects of miR-34a mimics and inhibitors on cell apoptosis rates analyzed by fluorescence-activated cell sorting. Cells were dually stained with Annexin V/PI and analyzed by flow cytometry to determine the population of cells in early and late apoptosis stages in the (A) negative control, (B) cells treated with miR-34a inhibitor and (C) cells treated with miR-34a mimics. Left lower quadrant, viable cells; right lower quadrant, early apoptotic cells; right upper quadrant, late apoptotic cells; left upper quadrant, necrotic cells. (D) Quantification of the total percentage of apoptotic cells in each group. **P<0.01, n=3 for each group. miR, microRNA; PI, propidium iodide.
Mentions: It is well known that miR-34a is an important component of the p53 tumor suppressor protein transcriptional network, which regulates cell proliferation and cell cycle progression. In order to investigate the biological effects of miR-34a in nerve cells, PC12 cells were transiently transfected with miR-34a mimics or miR-34a inhibitors, and the proportions of apoptotic cells were quantified using an Annexin V-FITC/PI dual staining assay. As shown in Fig. 2, after transfection, the total proportion of apoptotic cells in the miR-34a mimic group was significantly increased compared with that of the control group (P<0.01). The apoptotic rate of miR-34a inhibitor group was marginally lower than that of the control group, however, no significant difference was identified (P>0.05). The total apoptotic rates for control, miR-34a inhibitor and miR-34a mimic transfection groups were 12.2±1.06, 11.4±0.34 and 19.2±0.84%, respectively.

Bottom Line: The effect of miR‑34a in PC12 cells has not yet been reported.It was demonstrated that induction of miR‑34a promotes apoptosis and senescence, inhibits proliferation, and leads to marked alterations in SIRT1, Bcl‑12 and acetyl (ac)‑p53 expression.These data indicate that miR‑34a may be important in neuropathy.

View Article: PubMed Central - PubMed

Affiliation: Department of Rehabilitation Medicine, The First Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510080, P.R. China.

ABSTRACT
MicroRNA‑34a (miR‑34a) is a direct target of p53 and was reported to induce cell cycle arrest, apoptosis and senescence. Inhibition of the NAD‑dependent deacetylase sirtuin‑1 (SIRT1) by miR‑34a leads to an increase in acetylated p53, which promotes cell apoptosis. B‑cell lymphoma 2 (Bcl‑2) is also involved in apoptosis, and was originally characterized with respect to its role in controlling outer mitochondrial membrane integrity. The effect of miR‑34a in PC12 cells has not yet been reported. In the present study, it was hypothesized that Bcl‑2 and SIRT1 may be critical downstream targets of miR‑34a that participate in apoptosis induction. miR‑34a mimics and inhibitors were transfected into PC12 cells, and the apoptosis and proliferation rates were compared between groups. It was demonstrated that induction of miR‑34a promotes apoptosis and senescence, inhibits proliferation, and leads to marked alterations in SIRT1, Bcl‑12 and acetyl (ac)‑p53 expression. These data indicate that miR‑34a may be important in neuropathy.

Show MeSH
Related in: MedlinePlus