脐血干细胞表达转基因TNF-α联合溶瘤病毒对小鼠胃癌移植瘤抑制和杀伤作用  

Experimental study on mice gastric cancers treated by oncolytic virus with transgenic TNF-αexpressed in umbilical cord blood MSCs

作　　者：朱新红[1] 马贵亮[2] 毛伟征[2] 李谦[2] 战淑惠 周少飞[2] 林存智[3] 

机构地区：[1]青岛市市立医院国际门诊,山东青岛266071 [2]青岛市市立医院外科,山东青岛266071 [3]青岛大学附属医院呼吸肿瘤生物治疗科,山东青岛266003

出　　处：《中华肿瘤防治杂志》2015年第5期330-334,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30772542)

摘　　要：目的探讨脐血间质干细胞(umbilical cord blood mesenchymal stem cells,UCB-MSCs)表达转基因肿瘤坏死因子-α(tumor necrosis factor,TNF-α)联合溶瘤病毒-新城疫病毒(newcastle disease virus,NDV)抑制和杀伤胃癌的作用。方法构建带有人TNF-α基因的慢病毒载体,感染人脐血间质干细胞,获得重组慢病毒感染的脐血间质干细胞(MSC-TNF-α)。人胃癌细胞SGC-7901注射到裸鼠腹股沟皮下建立胃癌移植瘤裸鼠模型,荷瘤裸鼠分为MSC-TNF-α组、MSC-TNF-α联合溶瘤病毒组和生理盐水(NaCl)对照组,每组5只;对3组裸鼠瘤周分别注射MSC-TNF-α、MSC-TNF-α+溶瘤病毒和NaCl,隔日1次,共3次;第4周取材,观察注射前后瘤体生长情况。反转录PCR法测定3组胃癌组织中TNF-αmRNA表达;病理切片观察瘤体坏死面积及肿瘤细胞凋亡情况。结果 MSC-TNF-α组裸鼠肿瘤体积为(1.40±0.23)cm3,MSC-TNF-α+溶瘤病毒组为(0.94±0.28)cm3,NaCl组为(2.88±0.28)cm3,MSC-TNF-α+溶瘤病毒组与MSC-TNF-α组比较,差异有统计学意义,t=2.39,P=0.044;MSC-TNF-α组与NaCl组比较,差异有统计学意义,t=7.72,P<0.001。MSC-TNF-α组TNF-αmRNA相对表达量为5.96±0.87,NaCl组为1.71±0.75,两组比较差异有统计学意义,t=6.652,P<0.001;MSC-TNF-α+溶瘤病毒组为5.81±1.11,与NaCl组比较,差异有统计学意义,t=6.423,P<0.001;MSC-TNF-α组与MSC-TNF-α+溶瘤病毒组比较,差异无统计学意义,t=0.229,P=0.825。病理切片显示,MSC-TNF-α+溶瘤病毒组坏死面积最大。Hoechst凋亡检测示,MSC-TNF-α+溶瘤病毒组细胞凋亡率最高,两治疗组肝脏和肾脏功能检测均正常。结论转基因脐血间质干细胞旁分泌TNF-α联合溶瘤病毒对胃癌细胞具有明显的抑制杀伤作用。OBJECTIVE To investigate oncolytic virus-newcastle disease virus （NDV） with transgenic TNF-α car- ried by umbilical cord blood mesenchymal stem cells（UCB-MSCs） to treat gastric cancers in vivo. METHODS Lentiviral vectors carrying TNF-α gene had been constructed. Then UCB-MSCs were infected by recombinant Lentiviral and MSC-TNF-α was constructed. The SGC-7901 cells of human gastric cancer were injected in 15 nude mice subcutaneously in the groins. Three groups, 5 mice in each group, were injected around the tumors with MSC-TNF-α, MSC-TNF-α ＋ Oncolytic virus, or 0.9% NaCl solution in every other day, three times, and tumors＇ approximate volumes were measured until end of the 4th week. The expression of TNF-α mRNA was detected by reverse transcription PCR. Tumor tissues were sliced into section and the necrosis areas and cell apoptosis in the tumors observed. RESULTS The average tumor volumes of MSC-TNF-α group,MSC-TNF-α ＋ Oncolytic virus group and NaCl group were （1.40±0.23） cm3 , （0.94±0.28） cm3 and （2.88±0.28） cm^3 respectively. There were significant difference between MSC-TNF-α＋ Oneolytie virus group and MSC-TNF-α group 0=2.39,P= 0. 044）; There were significant difference between MSC-TNF-α group and NaCI group （t= 7.72,P〈0. 001）. TNF-α mRNA expression of MSC-TNF-α group,MSC-TNF-α ＋ Oncolytic virus group and NaCl group were 5.96 ± 0.87,5.81± 1.11 and 1.71±0.75 and the two treatment groups were different significantly compared with NaCl group （both P〈0. 001）. Sliced pathological sections of the tumor tissues showed that the tumor neerocytosis areas in the MSC-TNF-α ＋ Oneolytic virus group were the most compared with other two groups. MSC-TNF-α ＋ Onco- lyric virus group was found of significant apoptosis of tumor cells by Hoeehst staining. The liver function and kidney function showed normal in the two treatment groups. CONCLUSION MSC-TNF-α obviously targets and kills gastric cancer cells in nude mice as a bystander,and Oneolytic virus strength

关 键 词：胃肿瘤 肿瘤坏死因子-Α 脐血干细胞 溶瘤病毒 动物模型 裸鼠 

分 类 号：R735.2[医药卫生—肿瘤]