Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay
The Wnt signaling pathway has been associated with many essential cell processes. This study aims to examine the effects of Wnt signaling on proliferation of cultured HEK293T cells. Cells were incubated with Wnt3a, and the activation of the Wnt pathway was followed by analysis of the level of the β-...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2015-01-01
|
| Series: | International Journal of Cell Biology |
| Online Access: | http://dx.doi.org/10.1155/2015/928502 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849398813878386688 |
|---|---|
| author | Patricia Reischmann Johanna Fiebeck Nadine von der Weiden Oliver Müller |
| author_facet | Patricia Reischmann Johanna Fiebeck Nadine von der Weiden Oliver Müller |
| author_sort | Patricia Reischmann |
| collection | DOAJ |
| description | The Wnt signaling pathway has been associated with many essential cell processes. This study aims to examine the effects of Wnt signaling on proliferation of cultured HEK293T cells. Cells were incubated with Wnt3a, and the activation of the Wnt pathway was followed by analysis of the level of the β-catenin protein and of the expression levels of the target genes MYC and CCND1. The level of β-catenin protein increased up to fourfold. While the mRNA levels of c-Myc and cyclin D1 increased slightly, the protein levels increased up to a factor of 1.5. Remarkably, MTT and BrdU assays showed different results when measuring the proliferation rate of Wnt3a stimulated HEK293T cells. In the BrdU assays an increase of the proliferation rate could be detected, which correlated to the applied Wnt3a concentration. Oppositely, this correlation could not be shown in the MTT assays. The MTT results, which are based on the mitochondrial activity, were confirmed by analysis of the succinate dehydrogenase complex by immunofluorescence and by western blotting. Taken together, our study shows that Wnt3a activates proliferation of HEK293 cells. These effects can be detected by measuring DNA synthesis rather than by measuring changes of mitochondrial activity. |
| format | Article |
| id | doaj-art-9f2fe2dc8220465d8d1736a06a863028 |
| institution | Kabale University |
| issn | 1687-8876 1687-8884 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Cell Biology |
| spelling | doaj-art-9f2fe2dc8220465d8d1736a06a8630282025-08-20T03:38:30ZengWileyInternational Journal of Cell Biology1687-88761687-88842015-01-01201510.1155/2015/928502928502Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied AssayPatricia Reischmann0Johanna Fiebeck1Nadine von der Weiden2Oliver Müller3University of Applied Sciences Kaiserslautern, Molecular Biology, Amerikastraße 1, 66482 Zweibrücken, GermanyUniversity of Applied Sciences Kaiserslautern, Molecular Biology, Amerikastraße 1, 66482 Zweibrücken, GermanyPHAST GmbH, Entenmühlstraße 48, 66424 Homburg, GermanyUniversity of Applied Sciences Kaiserslautern, Molecular Biology, Amerikastraße 1, 66482 Zweibrücken, GermanyThe Wnt signaling pathway has been associated with many essential cell processes. This study aims to examine the effects of Wnt signaling on proliferation of cultured HEK293T cells. Cells were incubated with Wnt3a, and the activation of the Wnt pathway was followed by analysis of the level of the β-catenin protein and of the expression levels of the target genes MYC and CCND1. The level of β-catenin protein increased up to fourfold. While the mRNA levels of c-Myc and cyclin D1 increased slightly, the protein levels increased up to a factor of 1.5. Remarkably, MTT and BrdU assays showed different results when measuring the proliferation rate of Wnt3a stimulated HEK293T cells. In the BrdU assays an increase of the proliferation rate could be detected, which correlated to the applied Wnt3a concentration. Oppositely, this correlation could not be shown in the MTT assays. The MTT results, which are based on the mitochondrial activity, were confirmed by analysis of the succinate dehydrogenase complex by immunofluorescence and by western blotting. Taken together, our study shows that Wnt3a activates proliferation of HEK293 cells. These effects can be detected by measuring DNA synthesis rather than by measuring changes of mitochondrial activity.http://dx.doi.org/10.1155/2015/928502 |
| spellingShingle | Patricia Reischmann Johanna Fiebeck Nadine von der Weiden Oliver Müller Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay International Journal of Cell Biology |
| title | Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay |
| title_full | Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay |
| title_fullStr | Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay |
| title_full_unstemmed | Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay |
| title_short | Measured Effects of Wnt3a on Proliferation of HEK293T Cells Depend on the Applied Assay |
| title_sort | measured effects of wnt3a on proliferation of hek293t cells depend on the applied assay |
| url | http://dx.doi.org/10.1155/2015/928502 |
| work_keys_str_mv | AT patriciareischmann measuredeffectsofwnt3aonproliferationofhek293tcellsdependontheappliedassay AT johannafiebeck measuredeffectsofwnt3aonproliferationofhek293tcellsdependontheappliedassay AT nadinevonderweiden measuredeffectsofwnt3aonproliferationofhek293tcellsdependontheappliedassay AT olivermuller measuredeffectsofwnt3aonproliferationofhek293tcellsdependontheappliedassay |