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MicroRNA-199a-3p suppresses glioma cell proliferation by regulating the AKT/mTOR signaling pathway.

Shen L, Sun C, Li Y, Li X, Sun T, Liu C, Zhou Y, Du Z - Tumour Biol. (2015)

Bottom Line: The over-expression of miR-199a-3p might target mTOR and restrained cellular growth and proliferation but not invasive and apoptosis capability.Results indicated that cellular proliferation was inhibited to regulate the AKT/mTOR signaling pathway by elevating levels of miR-199a-3p.MiR-199a-3p in glioma cell lines has effects similar to the tumor suppressor gene on cellular proliferation via the AKT/mTOR signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Neurosurgery and Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu, 215006, People's Republic of China.

ABSTRACT
Glioma has been investigated for decades, but the prognosis remains poor because of rapid proliferation, its aggressive potential, and its resistance to chemotherapy or radiotherapy. The mammalian target of rapamycin (mTOR) is highly expressed and regulates cellular proliferation and cell growth. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene transcription and translation via up-regulating or down-regulating the levels of miRNAs. This study was conducted to explore the molecular functions of miR-199a-3p in glioma. We detected the expression of miR-199a-3p in glioma samples by quantitative PCR (qPCR). Then, we transfected the U87 and U251 cell lines with miR-199a-3p. Cellular proliferation, invasion, and apoptosis were assessed to explain the function of miR-199a-3p. PCR confirmed that the expression of miR-199a-3p was lower in glioma samples combined with normal brain tissues. The over-expression of miR-199a-3p might target mTOR and restrained cellular growth and proliferation but not invasive and apoptosis capability. Results indicated that cellular proliferation was inhibited to regulate the AKT/mTOR signaling pathway by elevating levels of miR-199a-3p. MiR-199a-3p in glioma cell lines has effects similar to the tumor suppressor gene on cellular proliferation via the AKT/mTOR signaling pathway.

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a Cellular proliferation was assessed after 72 h of incubation. Cells were treated with NVP-BEZ235 of 100 nmol/L for 24 h and then transfected further with miR-199a-3p (ampersand, P > 0.05), when compared with cells treated with NVP-BEZ235 only. b, c Immunofluorescence analysis showed that the expression of mTOR (asterisk, P < 0.05) and p-AKT (asterisks, P < 0.01) was apparently inhibited in cells by the up-regulation of miR-199a-3p
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Fig6: a Cellular proliferation was assessed after 72 h of incubation. Cells were treated with NVP-BEZ235 of 100 nmol/L for 24 h and then transfected further with miR-199a-3p (ampersand, P > 0.05), when compared with cells treated with NVP-BEZ235 only. b, c Immunofluorescence analysis showed that the expression of mTOR (asterisk, P < 0.05) and p-AKT (asterisks, P < 0.01) was apparently inhibited in cells by the up-regulation of miR-199a-3p

Mentions: Dates from the miRBase database and many studies [15, 17–19] had proved that mTOR was targeted by miR-199a-3p. AIQurashi et al.[20] reported that miR-199a-3p acted a role in tumor suppression by targeting mTORC1 and mTORC2. Moreover, the expression of mTOR in U87 and U51 cell lines obviously decreased after transfection with miR-199a-3p (P < 0.01; Fig. 2d). In addition, to estimate the effect of miR-199a-3p on AKT/mTOR, we also found mTOR protein levels and the phosphorylation of mTOR, AKT, p70S6K, and 4E-BP1 decreased in miR-199a-3p groups by Western blot analysis and immunofluorescence staining (P < 0.01; Figs. 5, 6, and 7). We further assessed the biological significance of miR-199a-3p in the regulation of mTOR expression in glioma cells. Cells treated with NVP-BEZ235 to inhibit mTOR activity and further transfected with miR-199a-3p did not have a significant difference in inhibiting cell proliferation (P > 0.05; Fig. 6a).Fig. 5


MicroRNA-199a-3p suppresses glioma cell proliferation by regulating the AKT/mTOR signaling pathway.

Shen L, Sun C, Li Y, Li X, Sun T, Liu C, Zhou Y, Du Z - Tumour Biol. (2015)

a Cellular proliferation was assessed after 72 h of incubation. Cells were treated with NVP-BEZ235 of 100 nmol/L for 24 h and then transfected further with miR-199a-3p (ampersand, P > 0.05), when compared with cells treated with NVP-BEZ235 only. b, c Immunofluorescence analysis showed that the expression of mTOR (asterisk, P < 0.05) and p-AKT (asterisks, P < 0.01) was apparently inhibited in cells by the up-regulation of miR-199a-3p
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4644202&req=5

Fig6: a Cellular proliferation was assessed after 72 h of incubation. Cells were treated with NVP-BEZ235 of 100 nmol/L for 24 h and then transfected further with miR-199a-3p (ampersand, P > 0.05), when compared with cells treated with NVP-BEZ235 only. b, c Immunofluorescence analysis showed that the expression of mTOR (asterisk, P < 0.05) and p-AKT (asterisks, P < 0.01) was apparently inhibited in cells by the up-regulation of miR-199a-3p
Mentions: Dates from the miRBase database and many studies [15, 17–19] had proved that mTOR was targeted by miR-199a-3p. AIQurashi et al.[20] reported that miR-199a-3p acted a role in tumor suppression by targeting mTORC1 and mTORC2. Moreover, the expression of mTOR in U87 and U51 cell lines obviously decreased after transfection with miR-199a-3p (P < 0.01; Fig. 2d). In addition, to estimate the effect of miR-199a-3p on AKT/mTOR, we also found mTOR protein levels and the phosphorylation of mTOR, AKT, p70S6K, and 4E-BP1 decreased in miR-199a-3p groups by Western blot analysis and immunofluorescence staining (P < 0.01; Figs. 5, 6, and 7). We further assessed the biological significance of miR-199a-3p in the regulation of mTOR expression in glioma cells. Cells treated with NVP-BEZ235 to inhibit mTOR activity and further transfected with miR-199a-3p did not have a significant difference in inhibiting cell proliferation (P > 0.05; Fig. 6a).Fig. 5

Bottom Line: The over-expression of miR-199a-3p might target mTOR and restrained cellular growth and proliferation but not invasive and apoptosis capability.Results indicated that cellular proliferation was inhibited to regulate the AKT/mTOR signaling pathway by elevating levels of miR-199a-3p.MiR-199a-3p in glioma cell lines has effects similar to the tumor suppressor gene on cellular proliferation via the AKT/mTOR signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Neurosurgery and Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu, 215006, People's Republic of China.

ABSTRACT
Glioma has been investigated for decades, but the prognosis remains poor because of rapid proliferation, its aggressive potential, and its resistance to chemotherapy or radiotherapy. The mammalian target of rapamycin (mTOR) is highly expressed and regulates cellular proliferation and cell growth. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene transcription and translation via up-regulating or down-regulating the levels of miRNAs. This study was conducted to explore the molecular functions of miR-199a-3p in glioma. We detected the expression of miR-199a-3p in glioma samples by quantitative PCR (qPCR). Then, we transfected the U87 and U251 cell lines with miR-199a-3p. Cellular proliferation, invasion, and apoptosis were assessed to explain the function of miR-199a-3p. PCR confirmed that the expression of miR-199a-3p was lower in glioma samples combined with normal brain tissues. The over-expression of miR-199a-3p might target mTOR and restrained cellular growth and proliferation but not invasive and apoptosis capability. Results indicated that cellular proliferation was inhibited to regulate the AKT/mTOR signaling pathway by elevating levels of miR-199a-3p. MiR-199a-3p in glioma cell lines has effects similar to the tumor suppressor gene on cellular proliferation via the AKT/mTOR signaling pathway.

Show MeSH
Related in: MedlinePlus