ABSTRACT
OBJECTIVE: The aim of this study was to explore the therapeutic effect of natural killer (NK) cells on human doxorubicin-sensitive and resistant breast adenocarcinoma. METHODS: Human doxorubicin-sensitive and resistant breast cancer cell lines (MCF-7 and MCF-7/ADR) were tagged with renilla luciferase (Rluc) (MCF-7/RC and MCF-7/ADR/RC). NK cells were tagged with enhanced firefly luciferase (effluc) using a recombinant retrovirus transfection (NKF). Expression of Rluc, effluc, and NK cell surface markers CD16, CD56 as well as death receptors, DR4 and DR5, were assessed by using flow cytometry. In vitro cytotoxic effect of NK to MCF-7 and MCF-7/ADR was measured and in vivo bioluminescence imaging was also performed to visualize MCF-7/RC, MCF-7/ADR, and NKF in an animal model. RESULTS: NK92-MI, MCF-7, and MCF-7/ADR cells were successfully labeled with Rluc or effluc. Both the target breast cancer cells (with Rluc) and therapeutic NK cells (with effluc) were noninvasively visualized in nude mice. Doxorubicin-resistant breast cancer cells (MCF-7/ADR) presented a higher expression of DR5 and were more sensitive to NK cells compared with doxorubicin-sensitive breast cancer cells (MCF-7). CONCLUSION: The results of present study suggest that NK cell therapy has a therapeutic effect on doxorubicin-sensitive and resistant breast cancer cells.
Subject(s)
Antibiotics, Antineoplastic/pharmacology , Breast Neoplasms/therapy , Breast/pathology , Doxorubicin/pharmacology , Drug Resistance, Neoplasm , Killer Cells, Natural/immunology , Killer Cells, Natural/transplantation , Animals , Breast/drug effects , Breast/immunology , Breast Neoplasms/immunology , Breast Neoplasms/pathology , Cell- and Tissue-Based Therapy , Female , Humans , MCF-7 Cells , Mice, Inbred BALB C , Mice, Nude , Optical Imaging , Receptors, Death Domain/analysis , Receptors, Death Domain/immunologyABSTRACT
OBJECTIVE: Apoptosis is frequently found in oral lichen planus (OLP) lesions, but the pathways leading to apoptosis are unknown. STUDY DESIGN: This study focused on analysis of caspase expression which is essential for apoptosis. Expression of caspases 2, 3, 8, 9, and 12 was studied in 70 biopsy samples from atrophic OLP to identify which cascade pathway, extrinsic or intrinsic, is of importance in apoptosis in OLP. RESULTS: Caspase-2 expression was present in every sample, and >70% of the epithelial cells were positive in 33% of the lesions. More than 70% of the epithelial cells expressed caspase-12 in 84% of the specimens. Caspase-8 expression was shown totally in 87% of the specimens. No caspase-3 expression was found in 57% of the samples, and caspase-9 expression was absent in the entire OLP specimen. CONCLUSIONS: The high frequency of intrinsic apoptotic pathway markers caspases 2 and 12 indicates intracellular stress in atrophic OLP epithelial cells.
Subject(s)
Apoptosis/physiology , Caspases, Effector/physiology , Caspases, Initiator/physiology , Lichen Planus, Oral/enzymology , Signal Transduction/physiology , Adult , Aged , Biopsy , Caspase 12/analysis , Caspase 2/analysis , Caspase 3/analysis , Caspase 8/analysis , Caspase 9/analysis , Caspases, Effector/analysis , Caspases, Initiator/analysis , Cysteine Endopeptidases/analysis , Epithelial Cells/enzymology , Female , Follow-Up Studies , Humans , Immunohistochemistry , Lichen Planus, Oral/pathology , Male , Middle Aged , Receptors, Death Domain/analysisABSTRACT
BACKGROUND: Following prolonged treatment with the non-steroidal anti-androgen bicalutamide (Casodex), LNCaP cells have become resistant to this drug. Previously, we found that the bicalutamide-refractory subline LNCaP-Bic acquires a growth advantage and does not respond to androgenic stimulation. In the present study, we have asked whether changes in response to the tumor-selective apoptosis inducer TNF-related apoptosis-inducing ligand (TRAIL) occur in LNCaP-Bic cells. METHODS: LNCaP and LNCaP-Bic cells were incubated with increasing concentrations of TRAIL and apoptosis rate was analyzed using FACS. Expression of death receptors (DR), adaptor protein Fas-associated death domain (FADD), members of the Bcl-2 family, and caspases were investigated by Western blot. RESULTS: The percentage of cells undergoing apoptosis was lower in LNCaP-Bic in comparison to LNCaP cells. There were no major differences in death receptor expression between control LNCaP and bicalutamide-selected cells. Surprisingly, treatment with TRAIL increased the levels of Bcl-2 by 50% in LNCaP-Bic cells. The ratio cleaved caspase/procaspase-8 was substantially lower in LNCaP-Bic cells. CONCLUSIONS: Reduced sensitivity to TRAIL-induced apoptosis is a novel mechanism relevant to resistance to bicalutamide in prostate cancer. Inability of TRAIL to cause programmed cell death might be caused by multiple perturbations in the TRAIL-signaling pathway.
