Limits...
MicroRNA-19a mediates gastric carcinoma cell proliferation through the activation of nuclear factor-κB.

Yang F, Wang H, Jiang Z, Hu A, Chu L, Sun Y, Han J - Mol Med Rep (2015)

Bottom Line: The present study predominantly focussed on the effects of microRNA (miR)‑19a on NF‑κB activation.The results of the western blot analysis demonstrated that the protein levels of p65 increased when the MGC‑803 cells were transfected with miR‑19a mimics.In addition, the downstream target genes of miR‑19a, including intercellular adhesion molecule, vascular cell adhesion molecule and monocyte chemoattractant protein‑1, were upregulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Research and Therapy Center, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China.

ABSTRACT
In gastric carcinoma, the nuclear factor‑κB (NF‑κB) signaling pathway is highly active, and the constitutive activation of NF‑κB prompts malignant cell proliferation. MicroRNAs are considered to be important mediators in the regulation of the NF‑κB signaling pathway. The present study predominantly focussed on the effects of microRNA (miR)‑19a on NF‑κB activation. Reverse transcription‑quantitative polymerase chain reaction was used to quantify the relative levels of miR‑19a in gastric carcinoma cells. MTT assays were used to determine the effect of miR‑19a on cellular proliferation. To detect the activation of NF‑κB, western blotting was performed to measure the protein levels of NF‑κB and the products of its downstream target genes. To define the target genes, luciferase reporter assays were used. miR‑19a was found to be markedly upregulated in gastric carcinoma cells. The overexpression of miR‑19a resulted in proliferation and enhanced migratory capabilities of the MGC‑803 gastric carcinoma cell line. The results of the western blot analysis demonstrated that the protein levels of p65 increased when the MGC‑803 cells were transfected with miR‑19a mimics. In addition, the downstream target genes of miR‑19a, including intercellular adhesion molecule, vascular cell adhesion molecule and monocyte chemoattractant protein‑1, were upregulated. The results of the luciferase assay indicated that IκB‑α was the target gene of miR‑19a. Therefore, the results of the present study suggested that miR‑19a enhances malignant gastric cell proliferation by constitutively activating the NF‑κB signaling pathway.

Show MeSH

Related in: MedlinePlus

Cell viability of the MGC-803 human gastric carcinoma cell line is increased by miR-19a. The MGC-803 cells were transfected with (A) miR-19a mimics or (B) miR-19a inhibitors and negative controls for 24, 48 or 72 h, respectively. The relative expression level of miR-19a was determined using reverse transcription-quantitative polymerase chain reaction and RNU48 was used an internal control. Cell viability was determined using an MTT assay. Data are presented as the mean ± standard error of three independent experiments. *P<0.05, vs. control. miR, microRNA; OD, optical density; NC, negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4581753&req=5

f2-mmr-12-04-5780: Cell viability of the MGC-803 human gastric carcinoma cell line is increased by miR-19a. The MGC-803 cells were transfected with (A) miR-19a mimics or (B) miR-19a inhibitors and negative controls for 24, 48 or 72 h, respectively. The relative expression level of miR-19a was determined using reverse transcription-quantitative polymerase chain reaction and RNU48 was used an internal control. Cell viability was determined using an MTT assay. Data are presented as the mean ± standard error of three independent experiments. *P<0.05, vs. control. miR, microRNA; OD, optical density; NC, negative control.

Mentions: To investigate the effect of miR-19a on MGC-803 cell viability, the MGC-803 cells were transfected with miR-19a mimics, inhibitors or negative controls for 24, 48 or 72 h, respectively. In this investigation, the mimics were analogues that enhanced the expression of miR-19a, whereas the inhibitors were analogues that reduced the expression of miR-19a. The MTT assay demonstrated that, when the miR-19a mimics were transfected into the MGC-803 human gastric carcinoma cell line, cell viability was significantly increased by 23 and 35% at 48 and 72 h, respectively (Fig. 2A). Conversely, when the expression of miR-19a was inhibited, cell viability was reduced by 17 and 36% at 48 and 72 h, respectively (Fig. 2B). These results indicated that miR-19a increased MGC-803 cell viability.


MicroRNA-19a mediates gastric carcinoma cell proliferation through the activation of nuclear factor-κB.

Yang F, Wang H, Jiang Z, Hu A, Chu L, Sun Y, Han J - Mol Med Rep (2015)

Cell viability of the MGC-803 human gastric carcinoma cell line is increased by miR-19a. The MGC-803 cells were transfected with (A) miR-19a mimics or (B) miR-19a inhibitors and negative controls for 24, 48 or 72 h, respectively. The relative expression level of miR-19a was determined using reverse transcription-quantitative polymerase chain reaction and RNU48 was used an internal control. Cell viability was determined using an MTT assay. Data are presented as the mean ± standard error of three independent experiments. *P<0.05, vs. control. miR, microRNA; OD, optical density; NC, negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581753&req=5

f2-mmr-12-04-5780: Cell viability of the MGC-803 human gastric carcinoma cell line is increased by miR-19a. The MGC-803 cells were transfected with (A) miR-19a mimics or (B) miR-19a inhibitors and negative controls for 24, 48 or 72 h, respectively. The relative expression level of miR-19a was determined using reverse transcription-quantitative polymerase chain reaction and RNU48 was used an internal control. Cell viability was determined using an MTT assay. Data are presented as the mean ± standard error of three independent experiments. *P<0.05, vs. control. miR, microRNA; OD, optical density; NC, negative control.
Mentions: To investigate the effect of miR-19a on MGC-803 cell viability, the MGC-803 cells were transfected with miR-19a mimics, inhibitors or negative controls for 24, 48 or 72 h, respectively. In this investigation, the mimics were analogues that enhanced the expression of miR-19a, whereas the inhibitors were analogues that reduced the expression of miR-19a. The MTT assay demonstrated that, when the miR-19a mimics were transfected into the MGC-803 human gastric carcinoma cell line, cell viability was significantly increased by 23 and 35% at 48 and 72 h, respectively (Fig. 2A). Conversely, when the expression of miR-19a was inhibited, cell viability was reduced by 17 and 36% at 48 and 72 h, respectively (Fig. 2B). These results indicated that miR-19a increased MGC-803 cell viability.

Bottom Line: The present study predominantly focussed on the effects of microRNA (miR)‑19a on NF‑κB activation.The results of the western blot analysis demonstrated that the protein levels of p65 increased when the MGC‑803 cells were transfected with miR‑19a mimics.In addition, the downstream target genes of miR‑19a, including intercellular adhesion molecule, vascular cell adhesion molecule and monocyte chemoattractant protein‑1, were upregulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Tumor Research and Therapy Center, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, P.R. China.

ABSTRACT
In gastric carcinoma, the nuclear factor‑κB (NF‑κB) signaling pathway is highly active, and the constitutive activation of NF‑κB prompts malignant cell proliferation. MicroRNAs are considered to be important mediators in the regulation of the NF‑κB signaling pathway. The present study predominantly focussed on the effects of microRNA (miR)‑19a on NF‑κB activation. Reverse transcription‑quantitative polymerase chain reaction was used to quantify the relative levels of miR‑19a in gastric carcinoma cells. MTT assays were used to determine the effect of miR‑19a on cellular proliferation. To detect the activation of NF‑κB, western blotting was performed to measure the protein levels of NF‑κB and the products of its downstream target genes. To define the target genes, luciferase reporter assays were used. miR‑19a was found to be markedly upregulated in gastric carcinoma cells. The overexpression of miR‑19a resulted in proliferation and enhanced migratory capabilities of the MGC‑803 gastric carcinoma cell line. The results of the western blot analysis demonstrated that the protein levels of p65 increased when the MGC‑803 cells were transfected with miR‑19a mimics. In addition, the downstream target genes of miR‑19a, including intercellular adhesion molecule, vascular cell adhesion molecule and monocyte chemoattractant protein‑1, were upregulated. The results of the luciferase assay indicated that IκB‑α was the target gene of miR‑19a. Therefore, the results of the present study suggested that miR‑19a enhances malignant gastric cell proliferation by constitutively activating the NF‑κB signaling pathway.

Show MeSH
Related in: MedlinePlus