CAJ | 학술논문

目的探讨IL-21转染的脐血造血干细胞(CD34~+UBSC·IL-21)对荷卵巢癌裸鼠的治疗作用.方法从脐血分离CD34~+造血干细胞,体外培养扩增后用于重组体pIRES2-IL-21-EGFP转染.以肿瘤大小、荷瘤鼠生存期判断CD34~+UBSC-IL-21对荷瘤裸鼠的治疗效应.以RT-PCR、免疫荧光、ELISA、Western blot、脾细胞增殖试验及免疫组化法分别鉴定CD34~+UBSC和肿瘤组织中IL-21的表达及活性.裸鼠脾细胞中NK细胞含量及脾细胞的杀伤效应、血清中IFN-γ和TNF-α水平分别用FCM与ELISA检测.结果 pIRES2-IL-21-EGFP成功转染CD34~+UBSC.CD34~+UBSC-IL-21能抑制肿瘤生长,延长荷瘤裸鼠生存期,治疗鼠肿瘤局部能表达IL-21、血清IFN-γ和TNF-α水平升高,NK细胞含量及NK细胞杀伤活性明显增强,与其他组相比,差异有统计学意义(P<0.01).结论转染IL-21的CD34~+UBSC有良好的抗裸鼠卵巢癌作用,该结果为临床使用UBSC为载体的基因治疗卵巢癌研究奠定了基础.
목적 탐토IL-21전염적제혈조혈간세포(CD34~+UBSC·IL-21)대하란소암라서적치료작용.방법 종제혈분리CD34~+조혈간세포,체외배양확증후용우중조체pIRES2-IL-21-EGFP전염.이종류대소、하류서생존기판단CD34~+UBSC-IL-21대하류라서적치료효응.이RT-PCR、면역형광、ELISA、Western blot、비세포증식시험급면역조화법분별감정CD34~+UBSC화종류조직중IL-21적표체급활성.라서비세포중NK세포함량급비세포적살상효응、혈청중IFN-γ화TNF-α수평분별용FCM여ELISA검측.결과 pIRES2-IL-21-EGFP성공전염CD34~+UBSC.CD34~+UBSC-IL-21능억제종류생장,연장하류라서생존기,치료서종류국부능표체IL-21、혈청IFN-γ화TNF-α수평승고,NK세포함량급NK세포살상활성명현증강,여기타조상비,차이유통계학의의(P<0.01).결론 전염IL-21적CD34~+UBSC유량호적항라서란소암작용,해결과위림상사용UBSC위재체적기인치료란소암연구전정료기출.
Objective To investigate the CD34~+ umbilical cord blood hematopoietic stem cells (CD34~+ UBSC) transfected with interleukin 21 (IL-21) against the ovarian cancer effect in tumor-bearing nude mice. Methods CD34~+ UBSC were obtained from the UBSC by a magnetically activated cell sorting technique and then transfected with recombinant plasmid plRES2-IL-21-EGFP after the CD34~+ UBSC were proliferated in vitro. The therapeutic effect was evaluated by the size of the tumor and the life span in nude mice treated with the CD34~+ UBSC-IL-21. The expression of IL-21 and its bioactivity in CD34~+ UBSC-IL-21 and in local neoplasitc tissues were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR), immune fluorescence technique, ELISA, Western blot, immunohistochemistry and splenocyte proliferative activity. The NK cell cytotoxicity and the numbers of NK cells, serum level of IFN-γ and TNF-αwere simultaneouly detected by FCM and ELISA, respectively. Results CD34~+ UBSC were cultured and transfected with pIRES2-IL-21-EGFP successfully. CD34~+ UBSC-IL-21 could inhibit the tumor growth and extended nude mice life span compared with other groups (P < 0.01). The expression of IL-21 in the neo-plastic tissue, serum level of IFN-γ and TNF-α , NK cell activity and the numbers of NK cells of mice origin and of human origin in splenocytes were increased significantly in the nude mice treated with CD34~+ UBSC-IL-21 compared with other groups(P <0.01). Conclusion The CD34~+ UBSC transfected with IL-21 have competent against ovarian cancer in tumor-bearing nude mice. The findings may establish a foundation for gene therapy of the ovarian cancer by CD34~+ UBSC-IL-21 in clinic application.