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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

TGFβR1 has a critical role in miR-181b-mediated cell growth, chemosensitivity to DDP and metastasis of NSCLC cells.(a) CCK analysis of the IC50 values of DDP in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (b) Flow cytometric analysis of apoptosis in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1 with DDP treatment. (c,d) Scratch, migration, invasion assay in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (e) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/ TGFβR1. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
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f5: TGFβR1 has a critical role in miR-181b-mediated cell growth, chemosensitivity to DDP and metastasis of NSCLC cells.(a) CCK analysis of the IC50 values of DDP in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (b) Flow cytometric analysis of apoptosis in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1 with DDP treatment. (c,d) Scratch, migration, invasion assay in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (e) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/ TGFβR1. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.

Mentions: We further investigated whether TGFβR1 was responsible for cell growth, chemosensitivity to DDP and metastasis of NSCLC cells after regulating miR-181b expression. Through the knock-down TGFβR1 in anti-miR-181b-A549 cells, cells were more sensitive to DDP, decreased migratory, invasive capability and inhibited EMT markers than control. Through transfection of TGFβR1 in miR-181b-A549/DDP cells, cells were more resistant to cisplatin, increased migratory, invasive capability and EMT markers expression than control (Fig. 5a–e).


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)

TGFβR1 has a critical role in miR-181b-mediated cell growth, chemosensitivity to DDP and metastasis of NSCLC cells.(a) CCK analysis of the IC50 values of DDP in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (b) Flow cytometric analysis of apoptosis in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1 with DDP treatment. (c,d) Scratch, migration, invasion assay in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (e) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/ TGFβR1. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
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f5: TGFβR1 has a critical role in miR-181b-mediated cell growth, chemosensitivity to DDP and metastasis of NSCLC cells.(a) CCK analysis of the IC50 values of DDP in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (b) Flow cytometric analysis of apoptosis in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1 with DDP treatment. (c,d) Scratch, migration, invasion assay in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/TGFβR1. (e) Western blot detection of E-cadherin, Vimentin, N-cadherin protein expression in A549/miR-181b inhibitors, A549/miR-181b inhibitors/si-TGFβR1, A549/DDP/miR-181b or A549/DDP/miR-181b/ TGFβR1. GAPDH was used as an internal control. *P < 0.05; **P < 0.01; ***P < 0.001.
Mentions: We further investigated whether TGFβR1 was responsible for cell growth, chemosensitivity to DDP and metastasis of NSCLC cells after regulating miR-181b expression. Through the knock-down TGFβR1 in anti-miR-181b-A549 cells, cells were more sensitive to DDP, decreased migratory, invasive capability and inhibited EMT markers than control. Through transfection of TGFβR1 in miR-181b-A549/DDP cells, cells were more resistant to cisplatin, increased migratory, invasive capability and EMT markers expression than control (Fig. 5a–e).

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