Targeting CXCR4 and FAK reverses doxorubicin resistance and suppresses invasion in non-small cell lung carcinoma.
BACKGROUND
Current high lung cancer mortality rates are mainly due to the occurrence of metastases and therapeutic resistance. Therefore, simultaneous targeting of these processes may be a valid approach for the treatment of this type of cancer. Here, we assessed relationships between CXC chemokine receptor type 4 (CXCR4) and focal adhesion kinase (FAK) gene expression levels and expression levels of the drug resistance-related genes ABCB1 and ABCC1, and tested the potential of CXCR4 and FAK inhibitors to reverse doxorubicin (DOX) resistance and to decrease the invasive capacity of non-small cell lung carcinoma (NSCLC) cells.


METHODS
qRT-PCR was used for gene expression analyses in primary lung tissue samples obtained from 30 NSCLC patients and the human NSCLC-derived cell lines NCI-H460, NCI-H460/R and COR-L23. MTT, flow cytometry, cell death and β-galactosidase activity assays were used to assess the in vitro impact of CXCR4 and FAK inhibitors on DOX sensitivity. In addition, invasion and gelatin degradation assays were used to assess the in vitro impact of the respective inhibitors on metastasis-related processes in combination with DOX treatment.


RESULTS
We found that ABCB1 over-expression was significantly associated with CXCR4 and FAK over-expression, whereas ABCC1 over-expression was associated with increased FAK expression. We also found that CXCR4 and FAK inhibitors strongly synergized with DOX in reducing cell viability, arresting the cell cycle in the S or G2/M phases and inducing senescence. Additionally, we found that DOX enhanced the anti-invasive potential of CXCR4 and FAK inhibitors by reducing gelatin degradation and invasion.


CONCLUSIONS
From our data we conclude that targeting of CXCR4 and FAK may overcome ABCB1 and ABCC1-dependent DOX resistance in NSCLC cells and that simultaneous treatment of these cells with DOX may potentiate the anti-invasive effects of CXCR4 and FAK inhibitors.
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia.
Miodrag
M
Dragoj
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia.
Zorica
Z
Milosevic
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia.
Jasna
J
Bankovic
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia.
Nikola
N
Tanic
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia.
Milica
M
Pesic
Institute for Biological Research "Sinisa Stankovic", Department of Neurobiology, University of Belgrade, Despota Stefana 142, Belgrade, 11060, Serbia. tijana.andjelkovic@ibiss.bg.ac.rs.
Tijana
T
Stankovic
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000096:biosynthesis / Q000235:genetics; D000631:Aminopyridines / Q000494:pharmacology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D001596:Benzylamines / Q000494:pharmacology; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000473:pathology*; D004317:Doxorubicin / Q000627:therapeutic use; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D051417:Focal Adhesion Kinase 1 / Q000037:antagonists & inhibitors* / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology*; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000096:biosynthesis / Q000235:genetics; D009361:Neoplasm Invasiveness / Q000235:genetics; D015363:Quinolones / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors* / Q000235:genetics; D013450:Sulfones / Q000494:pharmacology; D059467:Transcriptome
D018435:ATP Binding Cassette Transporter, Subfamily B; D000631:Aminopyridines; D001596:Benzylamines; D004317:Doxorubicin; D051417:Focal Adhesion Kinase 1; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D015363:Quinolones; D019718:Receptors, CXCR4; D013450:Sulfones; C521108:6-(4-(3-(methylsulfonyl)benzylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroquinolin-2(1H)-one; C513055:ABCB1 protein, human; C527276:CXCR4 protein, human; C525466:N,N'-di-2-pyridinyl-1,4-benzenedimethanamine; C496050:PTK2 protein, human; C421109:multidrug resistance-associated protein 1
CXCR4; DOX; FAK; Invasion; Non-small cell lung carcinoma; Resistance