miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1
Abstract Background miR-429 and TLN1 have been shown to affect the biological behaviours of many carcinomas. However, their effects in nasopharyngeal carcinoma (NPC) are not yet clear. Here, we investigated their regulatory relationships and effects on NPC cells. Methods TargetScan was used to predi...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-04-01
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| Series: | Cancer Cell International |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12935-019-0831-0 |
| _version_ | 1818108325724684288 |
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| author | Zhihui Wang Zhiquan Zhu Zhong Lin Youli Luo Zibin Liang Caibin Zhang Jianxu Chen Peijian Peng |
| author_facet | Zhihui Wang Zhiquan Zhu Zhong Lin Youli Luo Zibin Liang Caibin Zhang Jianxu Chen Peijian Peng |
| author_sort | Zhihui Wang |
| collection | DOAJ |
| description | Abstract Background miR-429 and TLN1 have been shown to affect the biological behaviours of many carcinomas. However, their effects in nasopharyngeal carcinoma (NPC) are not yet clear. Here, we investigated their regulatory relationships and effects on NPC cells. Methods TargetScan was used to predict the regulatory relationships of miR-429 and TLN1 in NPC cells. Then, Western blotting and quantitative real-time PCR (qPCR) were performed to examine TLN1 levels, and qPCR was used to determine miR-429 levels in NPC cell lines with different metastatic characteristics (5-8F, CNE-2, CNE-1, 6-10B and NP69), to investigate whether TLN1 and miR-429 are correlated with the metastatic characteristics of these cells. Next, we upregulated or downregulated miR-429 in 5-8F and 6-10B cells, which have different tumourigenicity and transferability, and examined TLN1 expression by western blotting and qPCR after transfection. QPCR was also performed to confirm successful transfection of miR-429 mimic into 5-8F and 6-10B cells. Dual luciferase reporter gene assay was performed to investigate whether miR-429 regulates TLN1 by binding to its 3′UTR. After transfection, Cell Counting Kit-8 (CCK8) and IncuCyte were used to examine the proliferation of these cells, and wound-healing assay, Transwell migration assay, and invasion assays were performed to investigate the changes in migration and invasion after transfection. Results Western blotting and qPCR analyses showed that the protein level of TLN1 was negatively correlated with miR-429 in NPC cell lines (P < 0.05), while the mRNA level showed no relation with miR429 expression (P > 0.05). In addition, cells with high transferability showed high TLN1 expression at the protein level, while miR429 expression showed the opposite trend (P < 0.05), but there were no differences at the mRNA level between the different cell lines. Overexpression of miR429 in 5-8F and 6-10B cells was accompanied by downregulation of TLN1 at the protein level (P < 0.05), while there were no significant differences at the mRNA level (P > 0.05). In addition, transferability, proliferation, and invasion were downregulated by miR429 overexpression (P < 0.05). However, dual-luciferase reporter gene assay indicated that TLN1 was not a direct target of miR-429. Conclusion This study showed that miR-429 functions as a tumour suppressor in NPC by downregulation of TLN1, although the relationship is not direct. |
| first_indexed | 2024-12-11T02:13:34Z |
| format | Article |
| id | doaj.art-c413a9cf1f434657803ba40bbac11600 |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-12-11T02:13:34Z |
| publishDate | 2019-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-c413a9cf1f434657803ba40bbac116002022-12-22T01:24:13ZengBMCCancer Cell International1475-28672019-04-0119111110.1186/s12935-019-0831-0miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1Zhihui Wang0Zhiquan Zhu1Zhong Lin2Youli Luo3Zibin Liang4Caibin Zhang5Jianxu Chen6Peijian Peng7Department of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Neurology, The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Hepatobiliary Surgery, The Fifth Affiliated Hospital of Sun Yat-sen UniversityDepartment of Thoracic Oncology, The Cancer Center of The Fifth Affiliated Hospital of Sun Yat-sen UniversityAbstract Background miR-429 and TLN1 have been shown to affect the biological behaviours of many carcinomas. However, their effects in nasopharyngeal carcinoma (NPC) are not yet clear. Here, we investigated their regulatory relationships and effects on NPC cells. Methods TargetScan was used to predict the regulatory relationships of miR-429 and TLN1 in NPC cells. Then, Western blotting and quantitative real-time PCR (qPCR) were performed to examine TLN1 levels, and qPCR was used to determine miR-429 levels in NPC cell lines with different metastatic characteristics (5-8F, CNE-2, CNE-1, 6-10B and NP69), to investigate whether TLN1 and miR-429 are correlated with the metastatic characteristics of these cells. Next, we upregulated or downregulated miR-429 in 5-8F and 6-10B cells, which have different tumourigenicity and transferability, and examined TLN1 expression by western blotting and qPCR after transfection. QPCR was also performed to confirm successful transfection of miR-429 mimic into 5-8F and 6-10B cells. Dual luciferase reporter gene assay was performed to investigate whether miR-429 regulates TLN1 by binding to its 3′UTR. After transfection, Cell Counting Kit-8 (CCK8) and IncuCyte were used to examine the proliferation of these cells, and wound-healing assay, Transwell migration assay, and invasion assays were performed to investigate the changes in migration and invasion after transfection. Results Western blotting and qPCR analyses showed that the protein level of TLN1 was negatively correlated with miR-429 in NPC cell lines (P < 0.05), while the mRNA level showed no relation with miR429 expression (P > 0.05). In addition, cells with high transferability showed high TLN1 expression at the protein level, while miR429 expression showed the opposite trend (P < 0.05), but there were no differences at the mRNA level between the different cell lines. Overexpression of miR429 in 5-8F and 6-10B cells was accompanied by downregulation of TLN1 at the protein level (P < 0.05), while there were no significant differences at the mRNA level (P > 0.05). In addition, transferability, proliferation, and invasion were downregulated by miR429 overexpression (P < 0.05). However, dual-luciferase reporter gene assay indicated that TLN1 was not a direct target of miR-429. Conclusion This study showed that miR-429 functions as a tumour suppressor in NPC by downregulation of TLN1, although the relationship is not direct.http://link.springer.com/article/10.1186/s12935-019-0831-0Nasopharyngeal carcinomamiR-429TLN1 |
| spellingShingle | Zhihui Wang Zhiquan Zhu Zhong Lin Youli Luo Zibin Liang Caibin Zhang Jianxu Chen Peijian Peng miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 Cancer Cell International Nasopharyngeal carcinoma miR-429 TLN1 |
| title | miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 |
| title_full | miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 |
| title_fullStr | miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 |
| title_full_unstemmed | miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 |
| title_short | miR-429 suppresses cell proliferation, migration and invasion in nasopharyngeal carcinoma by downregulation of TLN1 |
| title_sort | mir 429 suppresses cell proliferation migration and invasion in nasopharyngeal carcinoma by downregulation of tln1 |
| topic | Nasopharyngeal carcinoma miR-429 TLN1 |
| url | http://link.springer.com/article/10.1186/s12935-019-0831-0 |
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