中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
8期
544-548
,共5页
刘铮%马宏%史娟%张亚玺%刘彦信%郑德先
劉錚%馬宏%史娟%張亞璽%劉彥信%鄭德先
류쟁%마굉%사연%장아새%류언신%정덕선
干细胞%基因表达调控%癌,肝细胞%肿瘤坏死因子相关凋亡诱导配体%细胞治疗
榦細胞%基因錶達調控%癌,肝細胞%腫瘤壞死因子相關凋亡誘導配體%細胞治療
간세포%기인표체조공%암,간세포%종류배사인자상관조망유도배체%세포치료
Stem cells%Gene expression regulation%Carcinoma,hepatocellular%TRAIL%Cell therapy
目的 研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)在间充质干细胞中可控性表达及其对肿瘤细胞的杀伤活性,探索间充质干细胞用于肿瘤治疗的前景.方法 构建TRAIL可控性表达的腺病毒载体Ad-Tet-TRE-TRAIL,包装获得重组腺病毒后,感染小鼠间充质干细胞,免疫印迹法和酶联免疫吸附法测定间充质干细胞TRAIL可控性表达和分泌.四甲基偶氮唑盐(MTT)法测定TRAIL抑制肝癌细胞生长的能力.膜联蛋白(Annexin)-V/碘化丙啶(PI)染色和流式细胞术测定TRAIL对肝癌细胞的杀伤活性.结果 重组腺病毒Ad-Tet-TRE-TRAIL感染的小鼠间充质干细胞能够在强力霉素的控制下表达和分泌TRAIL,并显著抑制SMMC-7402人肝癌细胞生长,其机制是诱导肝癌细胞凋亡,结果显示SMMC-7402细胞在杀伤24、48 h后的存活率分别为66.5%±4.8%和42.9%±6.5%,而对照组(未加强力霉素)的细胞存活率分别为97.3%±2.2%和99.4%±4.7%.结论 间充质干细胞介导TRAIL可控性表达进而有效诱导肝癌细胞凋亡并抑制肝癌细胞生长,为肿瘤的细胞治疗提供了新的策略.
目的 研究腫瘤壞死因子相關凋亡誘導配體(TRAIL)在間充質榦細胞中可控性錶達及其對腫瘤細胞的殺傷活性,探索間充質榦細胞用于腫瘤治療的前景.方法 構建TRAIL可控性錶達的腺病毒載體Ad-Tet-TRE-TRAIL,包裝穫得重組腺病毒後,感染小鼠間充質榦細胞,免疫印跡法和酶聯免疫吸附法測定間充質榦細胞TRAIL可控性錶達和分泌.四甲基偶氮唑鹽(MTT)法測定TRAIL抑製肝癌細胞生長的能力.膜聯蛋白(Annexin)-V/碘化丙啶(PI)染色和流式細胞術測定TRAIL對肝癌細胞的殺傷活性.結果 重組腺病毒Ad-Tet-TRE-TRAIL感染的小鼠間充質榦細胞能夠在彊力黴素的控製下錶達和分泌TRAIL,併顯著抑製SMMC-7402人肝癌細胞生長,其機製是誘導肝癌細胞凋亡,結果顯示SMMC-7402細胞在殺傷24、48 h後的存活率分彆為66.5%±4.8%和42.9%±6.5%,而對照組(未加彊力黴素)的細胞存活率分彆為97.3%±2.2%和99.4%±4.7%.結論 間充質榦細胞介導TRAIL可控性錶達進而有效誘導肝癌細胞凋亡併抑製肝癌細胞生長,為腫瘤的細胞治療提供瞭新的策略.
목적 연구종류배사인자상관조망유도배체(TRAIL)재간충질간세포중가공성표체급기대종류세포적살상활성,탐색간충질간세포용우종류치료적전경.방법 구건TRAIL가공성표체적선병독재체Ad-Tet-TRE-TRAIL,포장획득중조선병독후,감염소서간충질간세포,면역인적법화매련면역흡부법측정간충질간세포TRAIL가공성표체화분비.사갑기우담서염(MTT)법측정TRAIL억제간암세포생장적능력.막련단백(Annexin)-V/전화병정(PI)염색화류식세포술측정TRAIL대간암세포적살상활성.결과 중조선병독Ad-Tet-TRE-TRAIL감염적소서간충질간세포능구재강력매소적공제하표체화분비TRAIL,병현저억제SMMC-7402인간암세포생장,기궤제시유도간암세포조망,결과현시SMMC-7402세포재살상24、48 h후적존활솔분별위66.5%±4.8%화42.9%±6.5%,이대조조(미가강력매소)적세포존활솔분별위97.3%±2.2%화99.4%±4.7%.결론 간충질간세포개도TRAIL가공성표체진이유효유도간암세포조망병억제간암세포생장,위종류적세포치료제공료신적책략.
Objective To study the controllabe expression of soluble tumor necrosis factor-related apoptosis-iuducing ligand (TRAIL) in mesenchymal stem cells and evaluate its potential tumoricidal effects in cancer therapy. Methods The controllable TRAIL expression vector of Ad-Tet-TRE-TRAIL was established in an adenovirus vector for transfection into murine mesenchymal stem cells. The controllable expression and secretion of TRAIL were detected by Western blot and enzyme-linked immunosorbent assay.The viability of hepatocellular carcinoma cells was determined by MTT assay. The tumoricidal activity of TRAIL was determined by Annexin-V/PI staining and flow cytometry. Results The murine expression model of TRAIL was successfully established in the presence of doxycycline. The secreted TRAIL in cell culture medium could efficaciously suppress the growth of human hepatocellular carcinoma SMMC-7402 by induced apoptosis. The cell viability of SMMC-7402 was 66. 5% ± 4. 8% and 42.9% ± 6. 5% at post-treatment versus 97.3% ±2. 2% and 99. 4% ±4. 7% in the control group at 24 h and 48 h. Conclusion The controllable TRAIL expression mediated by mesenchymal stem cells kills human hepatocellular carcinoma cells effectively. And it may provide a novel therapeutic strategy for hepatocellular carcinoma.
