LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a
Abstract Backgrounds Colorectal carcinoma (CRC) is a common malignant tumor. Increasing evidences indicated that CRC showed a resistance to 5‐fluorouracil (5‐FU) and further resulted in a poor prognosis. In this study, we aim to investigate the effect of long noncoding RNA nuclear paraspeckle assemb...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2020-02-01
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| Series: | Cancer Medicine |
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| Online Access: | https://doi.org/10.1002/cam4.2746 |
| _version_ | 1798023168387973120 |
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| author | Fen Liu Fei‐Yan Ai De‐Cai Zhang Li Tian Zhen‐Yun Yang Shao‐Jun Liu |
| author_facet | Fen Liu Fei‐Yan Ai De‐Cai Zhang Li Tian Zhen‐Yun Yang Shao‐Jun Liu |
| author_sort | Fen Liu |
| collection | DOAJ |
| description | Abstract Backgrounds Colorectal carcinoma (CRC) is a common malignant tumor. Increasing evidences indicated that CRC showed a resistance to 5‐fluorouracil (5‐FU) and further resulted in a poor prognosis. In this study, we aim to investigate the effect of long noncoding RNA nuclear paraspeckle assembly transcript 1 (LncRNA NEAT1) on cell viability, sensitivity to 5‐FU, and autophagy of CRC cell lines. Methods MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐Htetrazolium bromide) was used to detect cell viability, immunofluorescent staining was used to detect autophagy puncta, and luciferase reporter system was used to determine binding ability between miR‐34a and NEAT1 or putative targets. Additionally, indicated mRNAs and protein expressions were determined by qRT‐PCR or western blotting, respectively. Results We found that NEAT1 expression was increased in CRC tissues and cells, which showed a negative correlation with miR‐34a expression. In addition, NEAT1 knockdown noticeably inhibited the proliferation of CRC cells and enhanced 5‐FU sensitivity. It revealed that NEAT1 knockdown suppressed the LC3 puncta and the expressions of Beclin‐1, ULK1, and ratio of LC3II/I. Overexpression of miR‐34a showed similar trends with NEAT1 knockdown. miR‐34a was validated to target the putative binding sites in 3′‐UTR of HMGB1, ATG9A, and ATG4B, which are involved in the activation of autophagy. Inhibition of miR‐34a or overexpression of HMGB1 could effectively reverse elevated 5‐FU sensitivity upon NEAT1 knockdown. In addition, 3‐MA reversed NEAT1 overexpression‐induced resistance in HT29 cells. Conclusion These findings indicate that LncRNA NEAT1 could target miR‐34a and promote autophagy to facilitate 5‐FU chemoresistance in CRC. |
| first_indexed | 2024-04-11T17:43:08Z |
| format | Article |
| id | doaj.art-2b3d264a5dab4d80acacdc457aafc64f |
| institution | Directory Open Access Journal |
| issn | 2045-7634 |
| language | English |
| last_indexed | 2024-04-11T17:43:08Z |
| publishDate | 2020-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Cancer Medicine |
| spelling | doaj.art-2b3d264a5dab4d80acacdc457aafc64f2022-12-22T04:11:27ZengWileyCancer Medicine2045-76342020-02-01931079109110.1002/cam4.2746LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34aFen Liu0Fei‐Yan Ai1De‐Cai Zhang2Li Tian3Zhen‐Yun Yang4Shao‐Jun Liu5Department of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaDepartment of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaDepartment of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaDepartment of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaDepartment of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaDepartment of Gastroenterology The Third Xiangya Hospital of Central South University Changsha P.R. ChinaAbstract Backgrounds Colorectal carcinoma (CRC) is a common malignant tumor. Increasing evidences indicated that CRC showed a resistance to 5‐fluorouracil (5‐FU) and further resulted in a poor prognosis. In this study, we aim to investigate the effect of long noncoding RNA nuclear paraspeckle assembly transcript 1 (LncRNA NEAT1) on cell viability, sensitivity to 5‐FU, and autophagy of CRC cell lines. Methods MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐Htetrazolium bromide) was used to detect cell viability, immunofluorescent staining was used to detect autophagy puncta, and luciferase reporter system was used to determine binding ability between miR‐34a and NEAT1 or putative targets. Additionally, indicated mRNAs and protein expressions were determined by qRT‐PCR or western blotting, respectively. Results We found that NEAT1 expression was increased in CRC tissues and cells, which showed a negative correlation with miR‐34a expression. In addition, NEAT1 knockdown noticeably inhibited the proliferation of CRC cells and enhanced 5‐FU sensitivity. It revealed that NEAT1 knockdown suppressed the LC3 puncta and the expressions of Beclin‐1, ULK1, and ratio of LC3II/I. Overexpression of miR‐34a showed similar trends with NEAT1 knockdown. miR‐34a was validated to target the putative binding sites in 3′‐UTR of HMGB1, ATG9A, and ATG4B, which are involved in the activation of autophagy. Inhibition of miR‐34a or overexpression of HMGB1 could effectively reverse elevated 5‐FU sensitivity upon NEAT1 knockdown. In addition, 3‐MA reversed NEAT1 overexpression‐induced resistance in HT29 cells. Conclusion These findings indicate that LncRNA NEAT1 could target miR‐34a and promote autophagy to facilitate 5‐FU chemoresistance in CRC.https://doi.org/10.1002/cam4.2746autophagycolorectal carcinomaHMGB1LncRNA NEAT1miR‐34a |
| spellingShingle | Fen Liu Fei‐Yan Ai De‐Cai Zhang Li Tian Zhen‐Yun Yang Shao‐Jun Liu LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a Cancer Medicine autophagy colorectal carcinoma HMGB1 LncRNA NEAT1 miR‐34a |
| title | LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a |
| title_full | LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a |
| title_fullStr | LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a |
| title_full_unstemmed | LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a |
| title_short | LncRNA NEAT1 knockdown attenuates autophagy to elevate 5‐FU sensitivity in colorectal cancer via targeting miR‐34a |
| title_sort | lncrna neat1 knockdown attenuates autophagy to elevate 5 fu sensitivity in colorectal cancer via targeting mir 34a |
| topic | autophagy colorectal carcinoma HMGB1 LncRNA NEAT1 miR‐34a |
| url | https://doi.org/10.1002/cam4.2746 |
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