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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

Expression of miR-181b in NSCLC tissues is negatively correlated with TGFβR1 expression and responses of NSCLC patients to DDP-based chemotherapy.(a) qRT-PCR detection of relative miR-181b expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of miR-181b was normalized to U6 RNA. (b) qRT-PCR detection of relative TGFβR1 mRNA expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of TGFβR1 was normalized to GAPDH. (c) Immunostaining of TGFβR1 protein expression in sensitive and insensitive NSCLC tissues. (d) The OS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (e) The DFS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (f) A statistically significant inverse correlation between miR-181b and TGFβR1 mRNA levels in 38 cases of NSCLC tissues.
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f7: Expression of miR-181b in NSCLC tissues is negatively correlated with TGFβR1 expression and responses of NSCLC patients to DDP-based chemotherapy.(a) qRT-PCR detection of relative miR-181b expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of miR-181b was normalized to U6 RNA. (b) qRT-PCR detection of relative TGFβR1 mRNA expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of TGFβR1 was normalized to GAPDH. (c) Immunostaining of TGFβR1 protein expression in sensitive and insensitive NSCLC tissues. (d) The OS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (e) The DFS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (f) A statistically significant inverse correlation between miR-181b and TGFβR1 mRNA levels in 38 cases of NSCLC tissues.

Mentions: To investigate the correlation between miR-181b/TGFβR1 dysregulation and response to DDP and prognosis of patients, the expression of miR-181b and TGFβR1 mRNA was detected in a total of 38 NSCLC patients who received surgery, primary culture and subsequent MTT assay. Based on the patient’s response to DDP, they were divided into two groups: IR > 30% as sensitive; and IR≤30% as resistant. First, qPCR was used to detect the expression of miR-181b and TGFβR1 mRNA expression, and we showed that the relative level of miR-181b expression in sensitive tissues (n = 20) was significantly higher than that in insensitive tissues (n = 18) (P < 0.05; Fig. 7a). However, the relative level of TGFβR1 mRNA expression in sensitive groups was significantly lower than that in insensitive groups (P < 0.001; Fig. 7b). Immunostaining of TGFβR1 protein expression indicated that the staining of TGFβR1 protein was stronger in the insensitive tissues than in the sensitive tissues (Fig. 7c).


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)

Expression of miR-181b in NSCLC tissues is negatively correlated with TGFβR1 expression and responses of NSCLC patients to DDP-based chemotherapy.(a) qRT-PCR detection of relative miR-181b expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of miR-181b was normalized to U6 RNA. (b) qRT-PCR detection of relative TGFβR1 mRNA expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of TGFβR1 was normalized to GAPDH. (c) Immunostaining of TGFβR1 protein expression in sensitive and insensitive NSCLC tissues. (d) The OS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (e) The DFS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (f) A statistically significant inverse correlation between miR-181b and TGFβR1 mRNA levels in 38 cases of NSCLC tissues.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f7: Expression of miR-181b in NSCLC tissues is negatively correlated with TGFβR1 expression and responses of NSCLC patients to DDP-based chemotherapy.(a) qRT-PCR detection of relative miR-181b expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of miR-181b was normalized to U6 RNA. (b) qRT-PCR detection of relative TGFβR1 mRNA expression in sensitive group (n = 20) and insensitive group (n = 18) NSCLC tissues. Abundance of TGFβR1 was normalized to GAPDH. (c) Immunostaining of TGFβR1 protein expression in sensitive and insensitive NSCLC tissues. (d) The OS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (e) The DFS curves for the high-miR-181b expression group (n = 21) and the low-miR-181b expression group (n = 17). (f) A statistically significant inverse correlation between miR-181b and TGFβR1 mRNA levels in 38 cases of NSCLC tissues.
Mentions: To investigate the correlation between miR-181b/TGFβR1 dysregulation and response to DDP and prognosis of patients, the expression of miR-181b and TGFβR1 mRNA was detected in a total of 38 NSCLC patients who received surgery, primary culture and subsequent MTT assay. Based on the patient’s response to DDP, they were divided into two groups: IR > 30% as sensitive; and IR≤30% as resistant. First, qPCR was used to detect the expression of miR-181b and TGFβR1 mRNA expression, and we showed that the relative level of miR-181b expression in sensitive tissues (n = 20) was significantly higher than that in insensitive tissues (n = 18) (P < 0.05; Fig. 7a). However, the relative level of TGFβR1 mRNA expression in sensitive groups was significantly lower than that in insensitive groups (P < 0.001; Fig. 7b). Immunostaining of TGFβR1 protein expression indicated that the staining of TGFβR1 protein was stronger in the insensitive tissues than in the sensitive tissues (Fig. 7c).

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