Limits...
MiR-181b regulates cisplatin chemosensitivity and metastasis by targeting TGFβR1/Smad signaling pathway in NSCLC.

Wang X, Chen X, Meng Q, Jing H, Lu H, Yang Y, Cai L, Zhao Y - Sci Rep (2015)

Bottom Line: We found that miR-181b expression levels were lower in A549/DDP cells compared with A549 cells.In addition, miR-181b could inactivate the TGFβR1/Smad signaling pathway.We also observed that decreased miR-181b expression and increased TGFβR1 expression were significantly associated with chemosensitivity to DDP and tumor metastasis in NSCLC patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medical Oncology, Harbin medical University Cancer Hospital, Harbin, Heilongjiang Province, China.

ABSTRACT
MicroRNAs (miRNAs) have been identified as important post-transcriptional regulators involved in various biological and pathological processes of cells, but their underlying mechanisms in chemosensitivity and metastasis have not been fully elucidated. The objective of this study was to identify miR-181b and its mechanism in the chemosensitivity and metastasis of NSCLC. We found that miR-181b expression levels were lower in A549/DDP cells compared with A549 cells. Functional assays showed that the overexpression of miR-181b inhibited proliferation, enhanced chemosensitivity to DDP, attenuated migration and metastatic ability in NSCLC cell lines in vitro and in vivo. TGFβR1 was subsequently identified as a novel functional target of miR-181b. TGFβR1 knockdown revealed similar effects as that of ectopic miR-181b expression, whereas overexpression of TGFβR1 rescued the function of miR-181b-mediated growth, chemosensitivity and metastasis in NSCLC cells. In addition, miR-181b could inactivate the TGFβR1/Smad signaling pathway. We also observed that decreased miR-181b expression and increased TGFβR1 expression were significantly associated with chemosensitivity to DDP and tumor metastasis in NSCLC patients. Consequently, miR-181b functions as a tumor suppressor and has an important role in proliferation, chemosensitivity to DDP and metastasis of NSCLC by targeting TGFβR1/Smad signaling pathway.

No MeSH data available.


Related in: MedlinePlus

MiR-181b attenuates scratch, migration and invasion, modulates the epithelial- mesenchymal transition (EMT) of lung cancer cells.(a) Wound-healing assay in A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (b) Transwell migration and invasion assays for A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (c) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549 and A549/DDP after transduction with the miR-181b inhibitors, negative controls, or miR-181b mimics or miR-controls. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664936&req=5

f3: MiR-181b attenuates scratch, migration and invasion, modulates the epithelial- mesenchymal transition (EMT) of lung cancer cells.(a) Wound-healing assay in A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (b) Transwell migration and invasion assays for A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (c) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549 and A549/DDP after transduction with the miR-181b inhibitors, negative controls, or miR-181b mimics or miR-controls. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.

Mentions: We further studied the effects of miR-181b on cell migration and invasion using a scratch and transwell assay. A549/DDP cells transfected with miR-181b mimics showed less migratory ability than in miR-NC at 24 h after wound creation (Fig. 3a). The migratory and invasive abilities of A549/DDP cells were also reduced by 0.45 ± 0.03 and 0.26 ± 0.02 fold by over-expression of miR-181b compared with the capabilities of control cells (P < 0.05; P < 0.01; Fig. 3b). In contrast, A549 cells transfected with miR-181b inhibitors showed more migratory ability than in miR-NC at 24 h after wound creation. Migration and invasion were increased by 4.13 ± 0.24 and 3.60 ± 0.46 fold with miR-181b knockdown treatment in A549 cells (P < 0.05, P < 0.05; Fig. 3b). The same results were confirmed in H1650 cell (Fig. S3a,b).


MiR-181b regulates cisplatin chemosensitivity and metastasis by targeting TGFβR1/Smad signaling pathway in NSCLC.

Wang X, Chen X, Meng Q, Jing H, Lu H, Yang Y, Cai L, Zhao Y - Sci Rep (2015)

MiR-181b attenuates scratch, migration and invasion, modulates the epithelial- mesenchymal transition (EMT) of lung cancer cells.(a) Wound-healing assay in A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (b) Transwell migration and invasion assays for A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (c) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549 and A549/DDP after transduction with the miR-181b inhibitors, negative controls, or miR-181b mimics or miR-controls. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664936&req=5

f3: MiR-181b attenuates scratch, migration and invasion, modulates the epithelial- mesenchymal transition (EMT) of lung cancer cells.(a) Wound-healing assay in A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (b) Transwell migration and invasion assays for A549/DDP and A549 were determined after transduction with the miR-181b mimics, miR-controls, or miR-181b inhibitors or negative controls. (c) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549 and A549/DDP after transduction with the miR-181b inhibitors, negative controls, or miR-181b mimics or miR-controls. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
Mentions: We further studied the effects of miR-181b on cell migration and invasion using a scratch and transwell assay. A549/DDP cells transfected with miR-181b mimics showed less migratory ability than in miR-NC at 24 h after wound creation (Fig. 3a). The migratory and invasive abilities of A549/DDP cells were also reduced by 0.45 ± 0.03 and 0.26 ± 0.02 fold by over-expression of miR-181b compared with the capabilities of control cells (P < 0.05; P < 0.01; Fig. 3b). In contrast, A549 cells transfected with miR-181b inhibitors showed more migratory ability than in miR-NC at 24 h after wound creation. Migration and invasion were increased by 4.13 ± 0.24 and 3.60 ± 0.46 fold with miR-181b knockdown treatment in A549 cells (P < 0.05, P < 0.05; Fig. 3b). The same results were confirmed in H1650 cell (Fig. S3a,b).

Bottom Line: We found that miR-181b expression levels were lower in A549/DDP cells compared with A549 cells.In addition, miR-181b could inactivate the TGFβR1/Smad signaling pathway.We also observed that decreased miR-181b expression and increased TGFβR1 expression were significantly associated with chemosensitivity to DDP and tumor metastasis in NSCLC patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medical Oncology, Harbin medical University Cancer Hospital, Harbin, Heilongjiang Province, China.

ABSTRACT
MicroRNAs (miRNAs) have been identified as important post-transcriptional regulators involved in various biological and pathological processes of cells, but their underlying mechanisms in chemosensitivity and metastasis have not been fully elucidated. The objective of this study was to identify miR-181b and its mechanism in the chemosensitivity and metastasis of NSCLC. We found that miR-181b expression levels were lower in A549/DDP cells compared with A549 cells. Functional assays showed that the overexpression of miR-181b inhibited proliferation, enhanced chemosensitivity to DDP, attenuated migration and metastatic ability in NSCLC cell lines in vitro and in vivo. TGFβR1 was subsequently identified as a novel functional target of miR-181b. TGFβR1 knockdown revealed similar effects as that of ectopic miR-181b expression, whereas overexpression of TGFβR1 rescued the function of miR-181b-mediated growth, chemosensitivity and metastasis in NSCLC cells. In addition, miR-181b could inactivate the TGFβR1/Smad signaling pathway. We also observed that decreased miR-181b expression and increased TGFβR1 expression were significantly associated with chemosensitivity to DDP and tumor metastasis in NSCLC patients. Consequently, miR-181b functions as a tumor suppressor and has an important role in proliferation, chemosensitivity to DDP and metastasis of NSCLC by targeting TGFβR1/Smad signaling pathway.

No MeSH data available.


Related in: MedlinePlus