Basic Research - Cancer(구연) (E-004)

carboxyl esterase 과발현 hTERT 불멸화 인간지방줄기세포 병합치료에 의한 항암제 이리노테칸 거세저항성전립선암 성장억제 촉진
¹순천향대학교 의과대학 비뇨의학교실, ²의학연구소
송윤섭¹, 김재헌¹, 김도경¹, 두승환¹, 양원재¹, 오은정¹, 윤철원 ², 이상훈²
Purpose: Effective chemotherapy has not yet to be developed for castration-resistant prostate cancer (CRPC). Cell-mediated enzyme prodrug therapy including combination of carboxyl esterase (CE) and chemotherapeutic agent irinotecan (CPT-11) could be a possible treatment option. This study evaluated rabbit CE-overexpressing hTERT-immortalized human adipose stem cells (hTERT-ADSC.CE) for the inhibition of tumor growth in CRPC.
Materials and Methods: The hERT-ADSC.CE cell line was established by transfection with a lentiviral vector (CLV-Ubic)-encoding rabbit CE gene. Gene overexpression was confirmed via qPCR and western blot. In vitro or in vivo studies were performed to determine the potential for selective migration toward cancer cells. To determine in vitro suicide effects of hERT-ADSC.CE, cell culture under various concentrations of CPT-11 (0.01–5μM) was performed, and to determine in vitro cytotoxic effect of hERT-ADSC.CE cells, co-culturing of PC3 and hERT-ADSC.CE cells was performed. For in vivo model, PC3 cells (1 x106 cells) were injected subcutaneously into the flanks of nude mice, and hERT-ADSC.CE cells were injected via intracadiac route followed by continuous treatment using CPT-11 for 2 weeks. Final change of tumor volume was measured and immunohistochemical analysis was performed.
Result: The directional migration of hTERT-ADSC.CE cells toward PC3 cells was significantly stimulated in vitro. The selective migration of hTERT-ADSC.CE toward induced prostate cancer of mice was shown in vivo using real time PCR. Chemoattractant ligands and receptors of hTERT-ADSC.CE were shown. In vitro suicide effect of hTERT-ADSC.CE under the treatment of CPT-11 showed higher cytotoxicity at the concentrations of 5 μM than 1 μM CPT-11 at 72 h (p<0.05). Analysis of the in vitro cytotoxicity of PC3 cells with hTERT-ADSC.CE under CPT-11 treatment, showed that cell viability of PC3 cells decreased in hTERT-ADSC.CE compared with the CPT-11 alone (p<0.05). In the in vivo study, the inhibitory effects of hTERT-ADSC.CE combined with CPT-11 were higher than those of CPT-11 monotherapy (p<0.05).
Conclusion: CE-overexpressing ADSC potentiated the inhibition of tumor growth in CRPC-bearing mice in the presence of CPT-11 pro-drugs. These results showed a possibility of therapeutic stem cells expressing CE genes as a new therapeutic strategy against CRPC.
keywords : Stem cells, prostate cancer, carboxyl esterase