Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells
Objective: Multi-drug resistance (MDR) is a controversial issue in traditional chemotherapy of aggressive cancers, including hepatocellular carcinoma. The major cause of MDR is suggested to be the aberrant activation of the main signaling pathways such as Wnt and nuclear factor kappa-light-chain-...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2016-02-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/web/journal/article/1230/download |
| _version_ | 1817989090341027840 |
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| author | Nasrin Shojaie Seyed Mahmood Ghaffari |
| author_facet | Nasrin Shojaie Seyed Mahmood Ghaffari |
| author_sort | Nasrin Shojaie |
| collection | DOAJ |
| description | Objective: Multi-drug resistance (MDR) is a controversial issue in traditional chemotherapy
of aggressive cancers, including hepatocellular carcinoma. The major cause
of MDR is suggested to be the aberrant activation of the main signaling pathways
such as Wnt and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-
κB) which have key roles in the maintenance of cancer stem cells (CSCs). Therefore,
the evaluation of their alterations could be essential in chemo-resistant cancers such
as Hepatocellular carcinoma. The main purpose of this study was to investigate the
alteration of the mentioned pathways in the chemotherapy resistant cancer cells by
assessing their major molecular parameters.
Materials and Methods: In this experimental study, methylthiazol tetrazolium (MTT)
assay, acridine orange/ethidium bromide (AO/EtBr) and Hoechst 33342 staining,
DNA fragmentation and colony formation methods were employed to investigate the
cytotoxic effects of methotrexate (MTX) and doxorubicin (DOX) on PLC/PRF/5 cells.
Moreover, the expression of 11 important genes involved in MDR was performed by
semi-quantitative reverse transcriptase-polymerase chain reacti on (RT-PCR).
Results: PLC/PRF/5 cells (Alexander) were sensitive to DOX and normally resistant
to MTX. In addition, the results obtained from RT-PCR analysis revealed that
β-catenin expression was significantly reduced and ABCG2 significantly overexpressed
4.85 and 3.34 times (P value<0.05) in DOX and MTX treated cells, respectively.
Furthermore, a considerable expression of HIF-1α and p65 were detected only
in MTX-resistant cells.
Conclusion: Anti-cancer drugs may have more than one target in tumor cells. They
not only participate in deregulation of Wnt but also alter NF-κB activation. Moreover,
HIF-1α was the only anti-apoptotic protein that was significantly induced in the chemoresistant
cells. |
| first_indexed | 2024-04-14T00:42:48Z |
| format | Article |
| id | doaj.art-cdb3531ece7e483d864eb71e3cb9b30e |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-04-14T00:42:48Z |
| publishDate | 2016-02-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-cdb3531ece7e483d864eb71e3cb9b30e2022-12-22T02:22:08ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142016-02-01174730739Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 CellsNasrin Shojaie0Seyed Mahmood Ghaffari1Biochemistry Group, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, IranBiochemistry Group, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, IranObjective: Multi-drug resistance (MDR) is a controversial issue in traditional chemotherapy of aggressive cancers, including hepatocellular carcinoma. The major cause of MDR is suggested to be the aberrant activation of the main signaling pathways such as Wnt and nuclear factor kappa-light-chain-enhancer of activated B cells (NF- κB) which have key roles in the maintenance of cancer stem cells (CSCs). Therefore, the evaluation of their alterations could be essential in chemo-resistant cancers such as Hepatocellular carcinoma. The main purpose of this study was to investigate the alteration of the mentioned pathways in the chemotherapy resistant cancer cells by assessing their major molecular parameters. Materials and Methods: In this experimental study, methylthiazol tetrazolium (MTT) assay, acridine orange/ethidium bromide (AO/EtBr) and Hoechst 33342 staining, DNA fragmentation and colony formation methods were employed to investigate the cytotoxic effects of methotrexate (MTX) and doxorubicin (DOX) on PLC/PRF/5 cells. Moreover, the expression of 11 important genes involved in MDR was performed by semi-quantitative reverse transcriptase-polymerase chain reacti on (RT-PCR). Results: PLC/PRF/5 cells (Alexander) were sensitive to DOX and normally resistant to MTX. In addition, the results obtained from RT-PCR analysis revealed that β-catenin expression was significantly reduced and ABCG2 significantly overexpressed 4.85 and 3.34 times (P value<0.05) in DOX and MTX treated cells, respectively. Furthermore, a considerable expression of HIF-1α and p65 were detected only in MTX-resistant cells. Conclusion: Anti-cancer drugs may have more than one target in tumor cells. They not only participate in deregulation of Wnt but also alter NF-κB activation. Moreover, HIF-1α was the only anti-apoptotic protein that was significantly induced in the chemoresistant cells.http://celljournal.org/web/journal/article/1230/downloadDoxorubicinMDRMethotrexateNF-κBWnt |
| spellingShingle | Nasrin Shojaie Seyed Mahmood Ghaffari Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells Cell Journal Doxorubicin MDR Methotrexate NF-κB Wnt |
| title | Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells |
| title_full | Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells |
| title_fullStr | Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells |
| title_full_unstemmed | Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells |
| title_short | Simultaneous Analysis of Wnt and NF-κB Signaling Pathways in Doxorubicin Sensitive and Methotrexate Resistant PLC/ PRF/5 Cells |
| title_sort | simultaneous analysis of wnt and nf κb signaling pathways in doxorubicin sensitive and methotrexate resistant plc prf 5 cells |
| topic | Doxorubicin MDR Methotrexate NF-κB Wnt |
| url | http://celljournal.org/web/journal/article/1230/download |
| work_keys_str_mv | AT nasrinshojaie simultaneousanalysisofwntandnfkbsignalingpathwaysindoxorubicinsensitiveandmethotrexateresistantplcprf5cells AT seyedmahmoodghaffari simultaneousanalysisofwntandnfkbsignalingpathwaysindoxorubicinsensitiveandmethotrexateresistantplcprf5cells |