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Micro-ribonucleic acid 29b inhibits cell proliferation and invasion and enhances cell apoptosis and chemotherapy effects of cisplatin via targeting of DNMT3b and AKT3 in prostate cancer.

Yan B, Guo Q, Nan XX, Wang Z, Yin Z, Yi L, Wei YB, Gao YL, Zhou KQ, Yang JR - Onco Targets Ther (2015)

Bottom Line: After miR-29b mimics and inhibitors were successfully transfected into LNCaP, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was then used to investigate cell proliferation and cisplatin sensitivity of PCa cells.Based on bioinformatic methods, Western blot analysis, and dual-luciferase reporter assay, novel target genes of miR-29b were identified. miR-29b was downregulated in PCa tissues compared with matched adjacent nontumor tissues.Taken together, the results showed that miR-29b plays a tumor-suppressive role in PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Second Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT

Background: Micro-ribonucleic acids (miRNAs) are crucial regulators in malignant tumors. miRNA-29b (miR-29b) has been identified as a tumor suppressor in prostate cancer (PCa). However, very few studies have investigated the effects of miR-29b in PCa, especially the mechanism and its association with chemotherapy. Our study aimed to explore the role and mechanism of miR-29b in PCa.

Materials and methods: The expression levels of miR-29b were detected in ten clinical PCa specimens and four different PCa cell lines through quantitative real-time polymerase chain reaction. After miR-29b mimics and inhibitors were successfully transfected into LNCaP, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was then used to investigate cell proliferation and cisplatin sensitivity of PCa cells. Cell cycle, cell apoptosis, and cell invasion were detected via flow cytometry, annexin V-fluorescein isothiocyanate labeling, and transwell assay, respectively. Based on bioinformatic methods, Western blot analysis, and dual-luciferase reporter assay, novel target genes of miR-29b were identified.

Results: miR-29b was downregulated in PCa tissues compared with matched adjacent nontumor tissues. In the androgen-independent PCa cell line (LNCaP-AI), the expression of miR-29b was much lower than the androgen-dependent PCa cell line (LNCaP). Subsequent studies showed that forced expression of miR-29b inhibited cell proliferation and cell invasion and induced cell apoptosis in PCa. Upregulation of miR-29b also enhanced the chemosensitivity of PCa cells to cisplatin. Moreover, we identified DNMT3b and AKT3 as novel target genes of miR-29b in PCa.

Conclusion: Taken together, the results showed that miR-29b plays a tumor-suppressive role in PCa. It inhibits cell biological behavior and enhances the chemotherapy effects of cisplatin through its involvement in epigenetic regulation and PI3K/AKT pathway.

No MeSH data available.


Related in: MedlinePlus

Overexpression of miR-29b enhances chemosensitivity of cisplatin on prostate cancer cells. miR-29b targets DNMT3b and AKT3.Notes: (A) Compared with the blank-control (blank) and negative-control miR (Cont-miR) groups, the relative cell-growth rate in the miR-29b group was significantly lower along with action time, indicating that miR-29b inhibited cell proliferation of prostate cancer. Cell viability in the miR-29b group plus cisplatin was significantly lower than the miR plus cisplatin-control group and the miR-29b group, suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin for prostate cancer. (B) Western blotting of eight potential genes’ expression at the protein level in the blank, negative-control miRNA, miR-29b inhibitor-transfection, and miR-29b mimic-transfection groups. (C) The quantitative protein expression of DNMT3b, AKT3, and Mcl-1 were significantly increased in the miR-29b mimic-transfection group (Mimics) and decreased in the miR-29b inhibitor-transfection group (Inhibitors). *P<0.05.Abbreviation: miR, micro-ribonucleic acid.
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f3-ott-8-557: Overexpression of miR-29b enhances chemosensitivity of cisplatin on prostate cancer cells. miR-29b targets DNMT3b and AKT3.Notes: (A) Compared with the blank-control (blank) and negative-control miR (Cont-miR) groups, the relative cell-growth rate in the miR-29b group was significantly lower along with action time, indicating that miR-29b inhibited cell proliferation of prostate cancer. Cell viability in the miR-29b group plus cisplatin was significantly lower than the miR plus cisplatin-control group and the miR-29b group, suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin for prostate cancer. (B) Western blotting of eight potential genes’ expression at the protein level in the blank, negative-control miRNA, miR-29b inhibitor-transfection, and miR-29b mimic-transfection groups. (C) The quantitative protein expression of DNMT3b, AKT3, and Mcl-1 were significantly increased in the miR-29b mimic-transfection group (Mimics) and decreased in the miR-29b inhibitor-transfection group (Inhibitors). *P<0.05.Abbreviation: miR, micro-ribonucleic acid.

Mentions: There were five groups: the blank control group (Blank), LNCaP transfected with negative-control miRNA (Cont-miR), LNCaP transfected with negative-control miRNA plus cisplatin (Cont-miR + cisplatin), LNCaP transfected with miR-29b mimics (miR-29b), and LNCaP transfected with miR-29b mimics plus cisplatin (miR-29b + cisplatin). The LNCaP cell-growth rate in miR-29b was significant lower than in Blank and Cont-miR (P<0.05), indicating that miR-29b inhibited cell proliferation of PCa. The cell-growth rate in miR-29b + cisplatin was significantly lower than Cont-miR + cisplatin and miR-29b (P<0.05), suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin on PCa (Figure 3A).


Micro-ribonucleic acid 29b inhibits cell proliferation and invasion and enhances cell apoptosis and chemotherapy effects of cisplatin via targeting of DNMT3b and AKT3 in prostate cancer.

Yan B, Guo Q, Nan XX, Wang Z, Yin Z, Yi L, Wei YB, Gao YL, Zhou KQ, Yang JR - Onco Targets Ther (2015)

Overexpression of miR-29b enhances chemosensitivity of cisplatin on prostate cancer cells. miR-29b targets DNMT3b and AKT3.Notes: (A) Compared with the blank-control (blank) and negative-control miR (Cont-miR) groups, the relative cell-growth rate in the miR-29b group was significantly lower along with action time, indicating that miR-29b inhibited cell proliferation of prostate cancer. Cell viability in the miR-29b group plus cisplatin was significantly lower than the miR plus cisplatin-control group and the miR-29b group, suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin for prostate cancer. (B) Western blotting of eight potential genes’ expression at the protein level in the blank, negative-control miRNA, miR-29b inhibitor-transfection, and miR-29b mimic-transfection groups. (C) The quantitative protein expression of DNMT3b, AKT3, and Mcl-1 were significantly increased in the miR-29b mimic-transfection group (Mimics) and decreased in the miR-29b inhibitor-transfection group (Inhibitors). *P<0.05.Abbreviation: miR, micro-ribonucleic acid.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4356695&req=5

f3-ott-8-557: Overexpression of miR-29b enhances chemosensitivity of cisplatin on prostate cancer cells. miR-29b targets DNMT3b and AKT3.Notes: (A) Compared with the blank-control (blank) and negative-control miR (Cont-miR) groups, the relative cell-growth rate in the miR-29b group was significantly lower along with action time, indicating that miR-29b inhibited cell proliferation of prostate cancer. Cell viability in the miR-29b group plus cisplatin was significantly lower than the miR plus cisplatin-control group and the miR-29b group, suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin for prostate cancer. (B) Western blotting of eight potential genes’ expression at the protein level in the blank, negative-control miRNA, miR-29b inhibitor-transfection, and miR-29b mimic-transfection groups. (C) The quantitative protein expression of DNMT3b, AKT3, and Mcl-1 were significantly increased in the miR-29b mimic-transfection group (Mimics) and decreased in the miR-29b inhibitor-transfection group (Inhibitors). *P<0.05.Abbreviation: miR, micro-ribonucleic acid.
Mentions: There were five groups: the blank control group (Blank), LNCaP transfected with negative-control miRNA (Cont-miR), LNCaP transfected with negative-control miRNA plus cisplatin (Cont-miR + cisplatin), LNCaP transfected with miR-29b mimics (miR-29b), and LNCaP transfected with miR-29b mimics plus cisplatin (miR-29b + cisplatin). The LNCaP cell-growth rate in miR-29b was significant lower than in Blank and Cont-miR (P<0.05), indicating that miR-29b inhibited cell proliferation of PCa. The cell-growth rate in miR-29b + cisplatin was significantly lower than Cont-miR + cisplatin and miR-29b (P<0.05), suggesting that overexpression of miR-29b enhanced the chemotherapy of cisplatin on PCa (Figure 3A).

Bottom Line: After miR-29b mimics and inhibitors were successfully transfected into LNCaP, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was then used to investigate cell proliferation and cisplatin sensitivity of PCa cells.Based on bioinformatic methods, Western blot analysis, and dual-luciferase reporter assay, novel target genes of miR-29b were identified. miR-29b was downregulated in PCa tissues compared with matched adjacent nontumor tissues.Taken together, the results showed that miR-29b plays a tumor-suppressive role in PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Second Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China.

ABSTRACT

Background: Micro-ribonucleic acids (miRNAs) are crucial regulators in malignant tumors. miRNA-29b (miR-29b) has been identified as a tumor suppressor in prostate cancer (PCa). However, very few studies have investigated the effects of miR-29b in PCa, especially the mechanism and its association with chemotherapy. Our study aimed to explore the role and mechanism of miR-29b in PCa.

Materials and methods: The expression levels of miR-29b were detected in ten clinical PCa specimens and four different PCa cell lines through quantitative real-time polymerase chain reaction. After miR-29b mimics and inhibitors were successfully transfected into LNCaP, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was then used to investigate cell proliferation and cisplatin sensitivity of PCa cells. Cell cycle, cell apoptosis, and cell invasion were detected via flow cytometry, annexin V-fluorescein isothiocyanate labeling, and transwell assay, respectively. Based on bioinformatic methods, Western blot analysis, and dual-luciferase reporter assay, novel target genes of miR-29b were identified.

Results: miR-29b was downregulated in PCa tissues compared with matched adjacent nontumor tissues. In the androgen-independent PCa cell line (LNCaP-AI), the expression of miR-29b was much lower than the androgen-dependent PCa cell line (LNCaP). Subsequent studies showed that forced expression of miR-29b inhibited cell proliferation and cell invasion and induced cell apoptosis in PCa. Upregulation of miR-29b also enhanced the chemosensitivity of PCa cells to cisplatin. Moreover, we identified DNMT3b and AKT3 as novel target genes of miR-29b in PCa.

Conclusion: Taken together, the results showed that miR-29b plays a tumor-suppressive role in PCa. It inhibits cell biological behavior and enhances the chemotherapy effects of cisplatin through its involvement in epigenetic regulation and PI3K/AKT pathway.

No MeSH data available.


Related in: MedlinePlus