MicroRNA-143 suppresses gastric cancer cell growth and induces apoptosis by targeting COX-2
AIM: To investigate the function of microRNA-143 (miR-143) in gastric cancer and explore the target genes of miR-143.
METHODS: A quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to evaluate miR-143 expression in gastric cancer cell lines. After transfecting gastric cancer cells with miR-143-5p and miR-143-3p precursors, Alamar blue and apoptosis assays were used to measure the respective proliferation and apoptosis rates. Cyclooxygenase-2 (COX-2) expression was determined by real-time RT-PCR and Western blot assays after miR-143 transfection. Reporter plasmids were constructed, and a luciferase reporter assay was used to identify the miR-143 binding site on COX-2.
RESULTS: Both miR-143-5p and miR-143-3p were significantly downregulated in multiple gastric cancer cell lines. Forced miR-143-5p and miR-143-3p expression in gastric cancer cells produced a profound cytotoxic effect. MiR-145-5p transfection into gastric cancer cells resulted in a greater growth inhibitory effect (61.23% ± 3.16% vs. 46.58% ± 4.28%, P < 0.05 in the MKN-1 cell line) and a higher apoptosis rate (28.74% ± 1.93% vs. 22.13% ± 3.31%, P < 0.05 in the MKN-1 cell line) than miR-143-3p transfection. Further analysis indicated that COX-2 expression was potently suppressed by miR-143-5p but not by miR-143-3p. The activity of a luciferase reporter construct that contained the 3'-untranslated region (UTR) of COX-2 was downregulated by miR-143-5p (43.6% ± 4.86%, P < 0.01) but not by miR-143-3p. A mutation in the miR-145-5p binding site completely ablated the regulatory effect on luciferase activity, which suggests that there is a direct miR-145-5p binding site in the 3'-UTR of COX-2.
CONCLUSION: Both miR-143-5p and miR-143-3p function as anti-oncomirs in gastric cancer. However, miR-143-5p alone directly targets COX-2, and it exhibits a stronger tumor suppressive effect than miR-143-3p.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2013 |
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| Erschienen: |
2013 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:19 |
|---|---|
| Enthalten in: |
World journal of gastroenterology - 19(2013), 43 vom: 01., Seite 7758-65 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wu, Xiao-Li [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 03.07.2014 Date Revised 09.04.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.3748/wjg.v19.i43.7758 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM236277898 |
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| 024 | 7 | |a 10.3748/wjg.v19.i43.7758 |2 doi | |
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| 041 | |a eng | ||
| 100 | 1 | |a Wu, Xiao-Li |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a MicroRNA-143 suppresses gastric cancer cell growth and induces apoptosis by targeting COX-2 |
| 264 | 1 | |c 2013 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 03.07.2014 | ||
| 500 | |a Date Revised 09.04.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a AIM: To investigate the function of microRNA-143 (miR-143) in gastric cancer and explore the target genes of miR-143 | ||
| 520 | |a METHODS: A quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to evaluate miR-143 expression in gastric cancer cell lines. After transfecting gastric cancer cells with miR-143-5p and miR-143-3p precursors, Alamar blue and apoptosis assays were used to measure the respective proliferation and apoptosis rates. Cyclooxygenase-2 (COX-2) expression was determined by real-time RT-PCR and Western blot assays after miR-143 transfection. Reporter plasmids were constructed, and a luciferase reporter assay was used to identify the miR-143 binding site on COX-2 | ||
| 520 | |a RESULTS: Both miR-143-5p and miR-143-3p were significantly downregulated in multiple gastric cancer cell lines. Forced miR-143-5p and miR-143-3p expression in gastric cancer cells produced a profound cytotoxic effect. MiR-145-5p transfection into gastric cancer cells resulted in a greater growth inhibitory effect (61.23% ± 3.16% vs. 46.58% ± 4.28%, P < 0.05 in the MKN-1 cell line) and a higher apoptosis rate (28.74% ± 1.93% vs. 22.13% ± 3.31%, P < 0.05 in the MKN-1 cell line) than miR-143-3p transfection. Further analysis indicated that COX-2 expression was potently suppressed by miR-143-5p but not by miR-143-3p. The activity of a luciferase reporter construct that contained the 3'-untranslated region (UTR) of COX-2 was downregulated by miR-143-5p (43.6% ± 4.86%, P < 0.01) but not by miR-143-3p. A mutation in the miR-145-5p binding site completely ablated the regulatory effect on luciferase activity, which suggests that there is a direct miR-145-5p binding site in the 3'-UTR of COX-2 | ||
| 520 | |a CONCLUSION: Both miR-143-5p and miR-143-3p function as anti-oncomirs in gastric cancer. However, miR-143-5p alone directly targets COX-2, and it exhibits a stronger tumor suppressive effect than miR-143-3p | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Anti-oncomir | |
| 650 | 4 | |a Apoptosis | |
| 650 | 4 | |a Cyclooxygenase-2 | |
| 650 | 4 | |a Gastric cancer | |
| 650 | 4 | |a MicroRNA-143 | |
| 650 | 7 | |a 3' Untranslated Regions |2 NLM | |
| 650 | 7 | |a MIRN143 microRNA, human |2 NLM | |
| 650 | 7 | |a MicroRNAs |2 NLM | |
| 650 | 7 | |a Cyclooxygenase 2 |2 NLM | |
| 650 | 7 | |a EC 1.14.99.1 |2 NLM | |
| 650 | 7 | |a PTGS2 protein, human |2 NLM | |
| 650 | 7 | |a EC 1.14.99.1 |2 NLM | |
| 700 | 1 | |a Cheng, Bin |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Pei-Yuan |e verfasserin |4 aut | |
| 700 | 1 | |a Huang, Huan-Jun |e verfasserin |4 aut | |
| 700 | 1 | |a Zhao, Qiu |e verfasserin |4 aut | |
| 700 | 1 | |a Dan, Zi-Li |e verfasserin |4 aut | |
| 700 | 1 | |a Tian, De-An |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Peng |e verfasserin |4 aut | |
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