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Targeting of YAP1 by microRNA-15a and microRNA-16-1 exerts tumor suppressor function in gastric adenocarcinoma.

Kang W, Tong JH, Lung RW, Dong Y, Zhao J, Liang Q, Zhang L, Pan Y, Yang W, Pang JC, Cheng AS, Yu J, To KF - Mol. Cancer (2015)

Bottom Line: YAP1 was confirmed to be a functional target of miR-15a and miR-16-1 in GAC.YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in GAC cells.In clinical samples, YAP1 protein expression shows negative correlation with miR-15a and miR-16-1 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomical and Cellular Pathology, State Key Laboratory of Oncology in South China, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, SAR, PR China. weikang@cuhk.edu.hk.

ABSTRACT

Background: MicroRNAs (miRNAs) have been reported to play an important role in tumorigenesis. In this study, the role of miR-15a and miR-16-1 in gastric adenocarcinoma (GAC) was investigated.

Methods: The expression of miR-15a and miR-16-1 in cell lines and primary tumors was examined by miRNA qRT-PCR. Proliferative assays, colony formation, cell invasion and migration, flow cytometry analysis and in vivo study were performed by ectopic expression of miR-15a and miR-16-1. The putative target genes of miR-15a and miR-16-1 were explored by TargetScan and further validated.

Results: We found that miR-15a and miR-16-1 were down-regulated in GAC cell lines and primary tumor samples compared with normal gastric epithelium. Functional study demonstrated that ectopic expression of miR-15a and miR-16-1 suppressed cell proliferation, monolayer colony formation, invasion and migration, and xenograft formation in vivo. In addition, miR-15a and miR-16-1 induced G0/G1 cell cycle arrest which was further confirmed by Western blot and qRT-PCR of related cell cycle regulators. YAP1 was confirmed to be a functional target of miR-15a and miR-16-1 in GAC. YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in GAC cells. In clinical samples, YAP1 protein expression shows negative correlation with miR-15a and miR-16-1 expression.

Conclusion: In conclusion, targeting YAP1 by tumor suppressor miRNA miR-15a and miR-16-1 plays inhibitory effect and this might have a therapeutic potential in GAC.

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Related in: MedlinePlus

YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in NCI-N87 and MGC-803. (A) YAP1 re-expression increased cell proliferation compared with miR-15a or miR-16-1 alone (*, P < 0.05; **, P < 0.001). The SDs were achieved by the 575 nm absorbance readings in 6 wells of each item. (B) Monolayer colony formation assays revealed YAP1 re-expression abolished proliferation-inhibition of miR-15a and miR-16-1 partially (*, P < 0.05; **, P < 0.001). The representative colony formation pictures were shown in the bottom. The experiments was performed in triplicate wells to get SDs. (C) The suppressed cell invasion abilities was partially alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (*, P < 0.05; **, P < 0.001). The cells were counted in 3 random vision fields of each item to get SDs.
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Fig5: YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in NCI-N87 and MGC-803. (A) YAP1 re-expression increased cell proliferation compared with miR-15a or miR-16-1 alone (*, P < 0.05; **, P < 0.001). The SDs were achieved by the 575 nm absorbance readings in 6 wells of each item. (B) Monolayer colony formation assays revealed YAP1 re-expression abolished proliferation-inhibition of miR-15a and miR-16-1 partially (*, P < 0.05; **, P < 0.001). The representative colony formation pictures were shown in the bottom. The experiments was performed in triplicate wells to get SDs. (C) The suppressed cell invasion abilities was partially alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (*, P < 0.05; **, P < 0.001). The cells were counted in 3 random vision fields of each item to get SDs.

Mentions: YAP1 re-expression in rescuing the suppressive phenotypes of cancer cells by ectopic expression of miR-15a and miR-16-1 was investigated. Interestingly, the growth inhibitory phenotypes were partly alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (MTT proliferation assay, P < 0.05, Figure 5A; monolayer colony formation assay, P < 0.05, Figure 5B). The invasion-inhibitory effects of miR-15a and miR-16-1 were also partially diminished by YAP1 re-expression (P < 0.05, Figure 5C).Figure 5


Targeting of YAP1 by microRNA-15a and microRNA-16-1 exerts tumor suppressor function in gastric adenocarcinoma.

Kang W, Tong JH, Lung RW, Dong Y, Zhao J, Liang Q, Zhang L, Pan Y, Yang W, Pang JC, Cheng AS, Yu J, To KF - Mol. Cancer (2015)

YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in NCI-N87 and MGC-803. (A) YAP1 re-expression increased cell proliferation compared with miR-15a or miR-16-1 alone (*, P < 0.05; **, P < 0.001). The SDs were achieved by the 575 nm absorbance readings in 6 wells of each item. (B) Monolayer colony formation assays revealed YAP1 re-expression abolished proliferation-inhibition of miR-15a and miR-16-1 partially (*, P < 0.05; **, P < 0.001). The representative colony formation pictures were shown in the bottom. The experiments was performed in triplicate wells to get SDs. (C) The suppressed cell invasion abilities was partially alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (*, P < 0.05; **, P < 0.001). The cells were counted in 3 random vision fields of each item to get SDs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4342823&req=5

Fig5: YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in NCI-N87 and MGC-803. (A) YAP1 re-expression increased cell proliferation compared with miR-15a or miR-16-1 alone (*, P < 0.05; **, P < 0.001). The SDs were achieved by the 575 nm absorbance readings in 6 wells of each item. (B) Monolayer colony formation assays revealed YAP1 re-expression abolished proliferation-inhibition of miR-15a and miR-16-1 partially (*, P < 0.05; **, P < 0.001). The representative colony formation pictures were shown in the bottom. The experiments was performed in triplicate wells to get SDs. (C) The suppressed cell invasion abilities was partially alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (*, P < 0.05; **, P < 0.001). The cells were counted in 3 random vision fields of each item to get SDs.
Mentions: YAP1 re-expression in rescuing the suppressive phenotypes of cancer cells by ectopic expression of miR-15a and miR-16-1 was investigated. Interestingly, the growth inhibitory phenotypes were partly alleviated by YAP1 re-expression in NCI-N87 and MGC-803 cells (MTT proliferation assay, P < 0.05, Figure 5A; monolayer colony formation assay, P < 0.05, Figure 5B). The invasion-inhibitory effects of miR-15a and miR-16-1 were also partially diminished by YAP1 re-expression (P < 0.05, Figure 5C).Figure 5

Bottom Line: YAP1 was confirmed to be a functional target of miR-15a and miR-16-1 in GAC.YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in GAC cells.In clinical samples, YAP1 protein expression shows negative correlation with miR-15a and miR-16-1 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomical and Cellular Pathology, State Key Laboratory of Oncology in South China, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, SAR, PR China. weikang@cuhk.edu.hk.

ABSTRACT

Background: MicroRNAs (miRNAs) have been reported to play an important role in tumorigenesis. In this study, the role of miR-15a and miR-16-1 in gastric adenocarcinoma (GAC) was investigated.

Methods: The expression of miR-15a and miR-16-1 in cell lines and primary tumors was examined by miRNA qRT-PCR. Proliferative assays, colony formation, cell invasion and migration, flow cytometry analysis and in vivo study were performed by ectopic expression of miR-15a and miR-16-1. The putative target genes of miR-15a and miR-16-1 were explored by TargetScan and further validated.

Results: We found that miR-15a and miR-16-1 were down-regulated in GAC cell lines and primary tumor samples compared with normal gastric epithelium. Functional study demonstrated that ectopic expression of miR-15a and miR-16-1 suppressed cell proliferation, monolayer colony formation, invasion and migration, and xenograft formation in vivo. In addition, miR-15a and miR-16-1 induced G0/G1 cell cycle arrest which was further confirmed by Western blot and qRT-PCR of related cell cycle regulators. YAP1 was confirmed to be a functional target of miR-15a and miR-16-1 in GAC. YAP1 re-expression partly abrogated the inhibitory effect of miR-15a and miR-16-1 in GAC cells. In clinical samples, YAP1 protein expression shows negative correlation with miR-15a and miR-16-1 expression.

Conclusion: In conclusion, targeting YAP1 by tumor suppressor miRNA miR-15a and miR-16-1 plays inhibitory effect and this might have a therapeutic potential in GAC.

Show MeSH
Related in: MedlinePlus