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Cyclooxygenase-2, a Potential Therapeutic Target, Is Regulated by miR-101 in Esophageal Squamous Cell Carcinoma.

Shao Y, Li P, Zhu ST, Yue JP, Ji XJ, He Z, Ma D, Wang L, Wang YJ, Zong Y, Wu YD, Zhang ST - PLoS ONE (2015)

Bottom Line: The COX-2 and PEG2 levels were determined by western blot and enzyme-linked immunosorbent assays (ELISA).The antitumor effect of miR-101 was verified in a xenograft model.Furthermore, COX-2 was shown to be a target of miR-101.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Beijing Friendship Hospital, Capital Medical University, National Clinical Research Center for Digestive Disease, Beijing Key Laboratory for Precancerous Lesion of Digestive Diseases, Beijing, China.

ABSTRACT

Background & aims: Cyclooxygenase-2 (COX-2) is known to promote the carcinogenesis of esophageal squamous cell carcinoma (ESCC). There are no reports on whether microRNAs (miRNAs) regulate COX-2 expression in ESCC. This study investigated the effect of miR-101 on ESCC through modulating COX-2 expression in ESCC.

Methods: Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to quantify miR-101 expression in ESCC clinical tissues and cell lines. The effects of miR-101 on ESCC progression were evaluated by cell counting kit-8 (CCK8), transwell migration and invasion assays, as well as by flow cytometry. The COX-2 and PEG2 levels were determined by western blot and enzyme-linked immunosorbent assays (ELISA). The luciferase reporter assay was used to verify COX-2 as a direct target of miR-101. The anti-tumor activity of miR-101 in vivo was investigated in a xenograft nude mouse model of ESCC.

Results: Downregulation of miR-101 was confirmed through comparison of 30 pairs of ESCC tumor and adjacent normal tissues (P < 0.001), as well as in 11 ESCC cell lines and a human immortalized esophageal cell line (P < 0.001). Transfection of miR-101 in ESCC cell lines significantly suppressed cell proliferation, migration, and invasion (all P < 0.001). The antitumor effect of miR-101 was verified in a xenograft model. Furthermore, COX-2 was shown to be a target of miR-101.

Conclusions: Overexpression of miR-101 in ESCC inhibits proliferation and metastasis. Therefore, the miR-101/COX-2 pathway might be a therapeutic target in ESCC.

No MeSH data available.


Related in: MedlinePlus

Downregulation of miR-101 in tumor tissue specimens and cell lines of human ESCC.The expressions of miR-101 in 30 pairs of ESCC tumor tissues and corresponding normal tissues (A) and cell lines (B) were determined by quantitative real time RT-PCR as described in Materials and Methods. Results were calculated by 2-ΔΔCT method and shown as the mean value of three independent experiments. U6 was used as an internal control for data normalization of RT-PCR. Columns and error bars represent standard deviations from three independent measurements. ** P < 0.01; *** P < 0.001.
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pone.0140642.g001: Downregulation of miR-101 in tumor tissue specimens and cell lines of human ESCC.The expressions of miR-101 in 30 pairs of ESCC tumor tissues and corresponding normal tissues (A) and cell lines (B) were determined by quantitative real time RT-PCR as described in Materials and Methods. Results were calculated by 2-ΔΔCT method and shown as the mean value of three independent experiments. U6 was used as an internal control for data normalization of RT-PCR. Columns and error bars represent standard deviations from three independent measurements. ** P < 0.01; *** P < 0.001.

Mentions: 1. MiR-101 is down-regulated in ESCC. The expressions levels of miR-101 in clinical specimens of ESCC and the corresponding adjacent normal tissues obtained from 30 patients with ESCC, as well as in 11 ESCC cell lines and a human immortalized esophageal epithelia cell line (Het-1A) were determined. Compared to the adjacent normal tissues and Het-1A cell, the expressions levels of miR-101 were significantly downregulated in tumor tissues (Fig 1A) and 11 ESCC cell lines (Fig 1B).


Cyclooxygenase-2, a Potential Therapeutic Target, Is Regulated by miR-101 in Esophageal Squamous Cell Carcinoma.

Shao Y, Li P, Zhu ST, Yue JP, Ji XJ, He Z, Ma D, Wang L, Wang YJ, Zong Y, Wu YD, Zhang ST - PLoS ONE (2015)

Downregulation of miR-101 in tumor tissue specimens and cell lines of human ESCC.The expressions of miR-101 in 30 pairs of ESCC tumor tissues and corresponding normal tissues (A) and cell lines (B) were determined by quantitative real time RT-PCR as described in Materials and Methods. Results were calculated by 2-ΔΔCT method and shown as the mean value of three independent experiments. U6 was used as an internal control for data normalization of RT-PCR. Columns and error bars represent standard deviations from three independent measurements. ** P < 0.01; *** P < 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4640815&req=5

pone.0140642.g001: Downregulation of miR-101 in tumor tissue specimens and cell lines of human ESCC.The expressions of miR-101 in 30 pairs of ESCC tumor tissues and corresponding normal tissues (A) and cell lines (B) were determined by quantitative real time RT-PCR as described in Materials and Methods. Results were calculated by 2-ΔΔCT method and shown as the mean value of three independent experiments. U6 was used as an internal control for data normalization of RT-PCR. Columns and error bars represent standard deviations from three independent measurements. ** P < 0.01; *** P < 0.001.
Mentions: 1. MiR-101 is down-regulated in ESCC. The expressions levels of miR-101 in clinical specimens of ESCC and the corresponding adjacent normal tissues obtained from 30 patients with ESCC, as well as in 11 ESCC cell lines and a human immortalized esophageal epithelia cell line (Het-1A) were determined. Compared to the adjacent normal tissues and Het-1A cell, the expressions levels of miR-101 were significantly downregulated in tumor tissues (Fig 1A) and 11 ESCC cell lines (Fig 1B).

Bottom Line: The COX-2 and PEG2 levels were determined by western blot and enzyme-linked immunosorbent assays (ELISA).The antitumor effect of miR-101 was verified in a xenograft model.Furthermore, COX-2 was shown to be a target of miR-101.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Beijing Friendship Hospital, Capital Medical University, National Clinical Research Center for Digestive Disease, Beijing Key Laboratory for Precancerous Lesion of Digestive Diseases, Beijing, China.

ABSTRACT

Background & aims: Cyclooxygenase-2 (COX-2) is known to promote the carcinogenesis of esophageal squamous cell carcinoma (ESCC). There are no reports on whether microRNAs (miRNAs) regulate COX-2 expression in ESCC. This study investigated the effect of miR-101 on ESCC through modulating COX-2 expression in ESCC.

Methods: Real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to quantify miR-101 expression in ESCC clinical tissues and cell lines. The effects of miR-101 on ESCC progression were evaluated by cell counting kit-8 (CCK8), transwell migration and invasion assays, as well as by flow cytometry. The COX-2 and PEG2 levels were determined by western blot and enzyme-linked immunosorbent assays (ELISA). The luciferase reporter assay was used to verify COX-2 as a direct target of miR-101. The anti-tumor activity of miR-101 in vivo was investigated in a xenograft nude mouse model of ESCC.

Results: Downregulation of miR-101 was confirmed through comparison of 30 pairs of ESCC tumor and adjacent normal tissues (P < 0.001), as well as in 11 ESCC cell lines and a human immortalized esophageal cell line (P < 0.001). Transfection of miR-101 in ESCC cell lines significantly suppressed cell proliferation, migration, and invasion (all P < 0.001). The antitumor effect of miR-101 was verified in a xenograft model. Furthermore, COX-2 was shown to be a target of miR-101.

Conclusions: Overexpression of miR-101 in ESCC inhibits proliferation and metastasis. Therefore, the miR-101/COX-2 pathway might be a therapeutic target in ESCC.

No MeSH data available.


Related in: MedlinePlus