Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway
Objective. Rheumatoid arthritis (RA) multidrug resistance is associated with P-glycoprotein (P-gp) overexpression. We investigated the effects of methotrexate (MTX) alone and combined with 4-hydroperoxycyclophosphamide (4-HC) on P-gp expression in fibroblast-like synoviocytes (FLSs) from patients wi...
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| Language: | English |
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Wiley
2018-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2018/3619320 |
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| author | Kaili Qin Kailin Chen Wenpeng Zhao Xiangcong Zhao Jing Luo Qun Wang Chong Gao Xiaofeng Li Caihong Wang |
| author_facet | Kaili Qin Kailin Chen Wenpeng Zhao Xiangcong Zhao Jing Luo Qun Wang Chong Gao Xiaofeng Li Caihong Wang |
| author_sort | Kaili Qin |
| collection | DOAJ |
| description | Objective. Rheumatoid arthritis (RA) multidrug resistance is associated with P-glycoprotein (P-gp) overexpression. We investigated the effects of methotrexate (MTX) alone and combined with 4-hydroperoxycyclophosphamide (4-HC) on P-gp expression in fibroblast-like synoviocytes (FLSs) from patients with RA and examined the signaling pathway involved. Methods. RA-FLSs were treated with MTX, MTX + 4-HC, AG490 + MTX, or AG490 + MTX + 4-HC for 72 h. Proliferation inhibition rates were determined by MTT assay; P-gp expression was measured by flow cytometry and real-time polymerase chain reaction (RT-PCR); JAK2 and STAT3 were measured by RT-PCR and cell-based ELISA to assess STAT3 signaling. Results. MTX alone significantly induced P-gp expression and mRNA production in RA-FLSs. P-gp expression and mRNA levels were lower in the MTX + 4-HC group than in the MTX-alone group. In contrast to MTX, MTX + 4-HC reduced the STAT3 phosphorylation and downregulated JAK2 and STAT3 mRNA production. Inhibition of constitutively active STAT3 accompanied by 4-HC suppressed P-gp levels in RA-FLSs. The MTT assays revealed no significant differences in proliferation inhibition rates among groups. Conclusions. The increased anti-P-gp effect of MTX + 4-HC versus MTX alone in RA-FLSs was mediated via inhibition of the JAK2/STAT3 pathway and may have helped reverse MDR in refractory RA patients with high-P-gp levels. |
| format | Article |
| id | doaj-art-12f43bc1c04d44778cfaee4cd93e4d26 |
| institution | OA Journals |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-12f43bc1c04d44778cfaee4cd93e4d262025-08-20T02:01:42ZengWileyJournal of Immunology Research2314-88612314-71562018-01-01201810.1155/2018/36193203619320Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 PathwayKaili Qin0Kailin Chen1Wenpeng Zhao2Xiangcong Zhao3Jing Luo4Qun Wang5Chong Gao6Xiaofeng Li7Caihong Wang8Department of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaPathology, Joint Program in Transfusion Medicine, Brigham and Women’s Hospital/Children’s Hospital, Harvard Medical School, Boston, MA, USADepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaDepartment of Rheumatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, ChinaObjective. Rheumatoid arthritis (RA) multidrug resistance is associated with P-glycoprotein (P-gp) overexpression. We investigated the effects of methotrexate (MTX) alone and combined with 4-hydroperoxycyclophosphamide (4-HC) on P-gp expression in fibroblast-like synoviocytes (FLSs) from patients with RA and examined the signaling pathway involved. Methods. RA-FLSs were treated with MTX, MTX + 4-HC, AG490 + MTX, or AG490 + MTX + 4-HC for 72 h. Proliferation inhibition rates were determined by MTT assay; P-gp expression was measured by flow cytometry and real-time polymerase chain reaction (RT-PCR); JAK2 and STAT3 were measured by RT-PCR and cell-based ELISA to assess STAT3 signaling. Results. MTX alone significantly induced P-gp expression and mRNA production in RA-FLSs. P-gp expression and mRNA levels were lower in the MTX + 4-HC group than in the MTX-alone group. In contrast to MTX, MTX + 4-HC reduced the STAT3 phosphorylation and downregulated JAK2 and STAT3 mRNA production. Inhibition of constitutively active STAT3 accompanied by 4-HC suppressed P-gp levels in RA-FLSs. The MTT assays revealed no significant differences in proliferation inhibition rates among groups. Conclusions. The increased anti-P-gp effect of MTX + 4-HC versus MTX alone in RA-FLSs was mediated via inhibition of the JAK2/STAT3 pathway and may have helped reverse MDR in refractory RA patients with high-P-gp levels.http://dx.doi.org/10.1155/2018/3619320 |
| spellingShingle | Kaili Qin Kailin Chen Wenpeng Zhao Xiangcong Zhao Jing Luo Qun Wang Chong Gao Xiaofeng Li Caihong Wang Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway Journal of Immunology Research |
| title | Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway |
| title_full | Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway |
| title_fullStr | Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway |
| title_full_unstemmed | Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway |
| title_short | Methotrexate Combined with 4-Hydroperoxycyclophosphamide Downregulates Multidrug-Resistance P-Glycoprotein Expression Induced by Methotrexate in Rheumatoid Arthritis Fibroblast-Like Synoviocytes via the JAK2/STAT3 Pathway |
| title_sort | methotrexate combined with 4 hydroperoxycyclophosphamide downregulates multidrug resistance p glycoprotein expression induced by methotrexate in rheumatoid arthritis fibroblast like synoviocytes via the jak2 stat3 pathway |
| url | http://dx.doi.org/10.1155/2018/3619320 |
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