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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

MiR-181b regulates chemosensitivity to DDP and metastasis in vivo.Mice were treated with DDP (3.0 mg kg−1 body weight; i.p., thrice) or with 0.1 ml PBS (pH 7.4; i.p., thrice). (a) Representative images of tumor growth 28 days after injection using miR-181b agomir or miR-181b-NC agomir. Representative images of tumor volume growth curves (left). (b) Western blotting detect the expression of PARP and cleaved PARP in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. (c) Immunostaining of TGFβR1 protein expression in tumors developed from A549/DDP/ miR-181b agomir or A549/DDP/ miR-NC agomir cells treated with DDP or PBS. Up: H&E staining; middle: TUNEL assay; down: immunostaining. Original magnification: × 200. The bar graph is the apoptosis rate calculated of TUNEL assay. (d) Representative photos of mouse lungs and images of the histological inspection of mouse lungs for the presence of microscopic lesions at 7 weeks after tail vein injection with A549/DDP cells and injected subcutaneously miR-181b agomir or miR-181b-NC agomir. (e) qPCR and Western blotting detect the expression of miR-181b , TGFβR1 in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. Data were presented as mean ± s. d., (f) Hypothesized mechanisms of miR-181b regulate cisplatin-resistant and metastasis in NSCLC. *P < 0.05; **P < 0.01.
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f8: MiR-181b regulates chemosensitivity to DDP and metastasis in vivo.Mice were treated with DDP (3.0 mg kg−1 body weight; i.p., thrice) or with 0.1 ml PBS (pH 7.4; i.p., thrice). (a) Representative images of tumor growth 28 days after injection using miR-181b agomir or miR-181b-NC agomir. Representative images of tumor volume growth curves (left). (b) Western blotting detect the expression of PARP and cleaved PARP in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. (c) Immunostaining of TGFβR1 protein expression in tumors developed from A549/DDP/ miR-181b agomir or A549/DDP/ miR-NC agomir cells treated with DDP or PBS. Up: H&E staining; middle: TUNEL assay; down: immunostaining. Original magnification: × 200. The bar graph is the apoptosis rate calculated of TUNEL assay. (d) Representative photos of mouse lungs and images of the histological inspection of mouse lungs for the presence of microscopic lesions at 7 weeks after tail vein injection with A549/DDP cells and injected subcutaneously miR-181b agomir or miR-181b-NC agomir. (e) qPCR and Western blotting detect the expression of miR-181b , TGFβR1 in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. Data were presented as mean ± s. d., (f) Hypothesized mechanisms of miR-181b regulate cisplatin-resistant and metastasis in NSCLC. *P < 0.05; **P < 0.01.

Mentions: To explore whether miR-181b could influence lung cancer chemosensitivity to DDP in vivo, s.c. tumors were developed in nude mice followed by treatment with agomir-181b or agomir-NC. All Nude mice were treated with DDP. As shown in Fig. 8a, the tumors treated with agomir-181b grew significantly slower than those with agomir-NC after the treatment with DDP. At 35 days after inoculation, the average tumor volume of A549/DDP/agomir-181b groups was significantly lower than that of A549/DDP/agomir-NC following DDP treatment (P < 0.01). Agomir-181b with DDP increased the expression of cleaved PARP compared to NC-agomir with DDP (Fig. 8b). TUNEL assay showed that the apoptotic rate of tumors developed from A549/DDP/agomir-181b groups (22.33 ± 0.88%) was significantly higher than that of tumors developed from A549/DDP/agomir-NC groups (11.67 ± 0.88%) following DDP treatment (P < 0.01; Fig. 8b,c).


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 regulates chemosensitivity to DDP and metastasis in vivo.Mice were treated with DDP (3.0 mg kg−1 body weight; i.p., thrice) or with 0.1 ml PBS (pH 7.4; i.p., thrice). (a) Representative images of tumor growth 28 days after injection using miR-181b agomir or miR-181b-NC agomir. Representative images of tumor volume growth curves (left). (b) Western blotting detect the expression of PARP and cleaved PARP in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. (c) Immunostaining of TGFβR1 protein expression in tumors developed from A549/DDP/ miR-181b agomir or A549/DDP/ miR-NC agomir cells treated with DDP or PBS. Up: H&E staining; middle: TUNEL assay; down: immunostaining. Original magnification: × 200. The bar graph is the apoptosis rate calculated of TUNEL assay. (d) Representative photos of mouse lungs and images of the histological inspection of mouse lungs for the presence of microscopic lesions at 7 weeks after tail vein injection with A549/DDP cells and injected subcutaneously miR-181b agomir or miR-181b-NC agomir. (e) qPCR and Western blotting detect the expression of miR-181b , TGFβR1 in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. Data were presented as mean ± s. d., (f) Hypothesized mechanisms of miR-181b regulate cisplatin-resistant and metastasis in NSCLC. *P < 0.05; **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f8: MiR-181b regulates chemosensitivity to DDP and metastasis in vivo.Mice were treated with DDP (3.0 mg kg−1 body weight; i.p., thrice) or with 0.1 ml PBS (pH 7.4; i.p., thrice). (a) Representative images of tumor growth 28 days after injection using miR-181b agomir or miR-181b-NC agomir. Representative images of tumor volume growth curves (left). (b) Western blotting detect the expression of PARP and cleaved PARP in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. (c) Immunostaining of TGFβR1 protein expression in tumors developed from A549/DDP/ miR-181b agomir or A549/DDP/ miR-NC agomir cells treated with DDP or PBS. Up: H&E staining; middle: TUNEL assay; down: immunostaining. Original magnification: × 200. The bar graph is the apoptosis rate calculated of TUNEL assay. (d) Representative photos of mouse lungs and images of the histological inspection of mouse lungs for the presence of microscopic lesions at 7 weeks after tail vein injection with A549/DDP cells and injected subcutaneously miR-181b agomir or miR-181b-NC agomir. (e) qPCR and Western blotting detect the expression of miR-181b , TGFβR1 in tumors developed from miR-181b agomir or miR-181b-NC agomir treated. Data were presented as mean ± s. d., (f) Hypothesized mechanisms of miR-181b regulate cisplatin-resistant and metastasis in NSCLC. *P < 0.05; **P < 0.01.
Mentions: To explore whether miR-181b could influence lung cancer chemosensitivity to DDP in vivo, s.c. tumors were developed in nude mice followed by treatment with agomir-181b or agomir-NC. All Nude mice were treated with DDP. As shown in Fig. 8a, the tumors treated with agomir-181b grew significantly slower than those with agomir-NC after the treatment with DDP. At 35 days after inoculation, the average tumor volume of A549/DDP/agomir-181b groups was significantly lower than that of A549/DDP/agomir-NC following DDP treatment (P < 0.01). Agomir-181b with DDP increased the expression of cleaved PARP compared to NC-agomir with DDP (Fig. 8b). TUNEL assay showed that the apoptotic rate of tumors developed from A549/DDP/agomir-181b groups (22.33 ± 0.88%) was significantly higher than that of tumors developed from A549/DDP/agomir-NC groups (11.67 ± 0.88%) following DDP treatment (P < 0.01; Fig. 8b,c).

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