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目的:以脐血间质干细胞(umbilical cord blood mesenchyme stem cell,UCBMSC)作为肿瘤坏死因子-α(tumor necro-sis factor-alpha,TNF-α)载体,研究重组TNF-α慢病毒Lv-TNF-α感染的UCBMSC对胃癌移植瘤生长的抑制作用。方法: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。将表达TNF-α的重组慢病毒Lv-TNF-α和对照慢病毒Lv-EGFP分别感染UCBMSC后获得Lv-TNF-α感染的UCBMSC(UCBMSC-TNF-α)以及Lv-EGFP感染的UCBMSC(UCBMSC-EGFP)。荷瘤裸鼠随机分为3组:分别注射UCBMSC-TNF-α、UCBMSC-EGFP及生理盐水(NaCl),观察注射后瘤体生长情况,RT-PCR和ELISA方法分别测定各组胃癌组织中TNF-αmRNA和蛋白的表达;H-E染色观察瘤体的坏死情况。结果:成功建立SGC-7901胃癌细胞裸鼠移植瘤模型。治疗后三组荷瘤裸鼠肿瘤体积分别为(0.51±0.27)、(0.64±0.36)和(1.21±0.80)cm3,UCBMSC-TNF-α组肿瘤体积最小(F=3.88,P<0.05);RT-PCR法检测结果显示,3组荷瘤裸鼠肿瘤组织TNF-αmRNA分别为(1.92±0.12)、(1.21±0.26)、(0.81±0.22),UCBMSC-TNF-α组TNF-αmRNA表达量最大(F=54.82,P<0.01);ELISA法检测结果显示,3组荷瘤裸鼠肿瘤组织TNF-α蛋白表达分别为(148.29±3.76)、(78.22±6.24)、(42.80±3.02)pg/ml,MSC-TNF-α组表达量最大(F=694.54,P<0.01);H-E染色病理切片显示,UCBMSC-TNF-α治疗组肿瘤坏死面积最大。结论:TNF-α转基因UCBMSC可通过旁分泌TNF-α抑制胃癌的生长。
OBJECTIVE: To investigate the effect of recombinant TNF-α lentivirus Lv-TNF-α on the expression of tumor necrosis factor-α (TNF-α) in umbilical cord blood mesenchyme stem cells (UCBMSC) Inhibitory effect of α-infected UCBMSCs on the growth of gastric cancer xenografts. Methods: Human gastric cancer cell line SGC-7901 was injected into the groin of nude mice to establish a model of gastric cancer xenografts in nude mice. UCBMSC (UCBMSC-TNF-α) infected with Lv-TNF-α and UCBMSC (UCBMSC) infected with Lv-EGFP were infected with recombinant lentivirus Lv-TNF-α expressing TNF-α and control lentivirus Lv- -EGFP). The tumor-bearing nude mice were randomly divided into 3 groups: UCBMSC-TNF-α, UCBMSC-EGFP and normal saline (NaCl) were injected respectively to observe the growth of the tumor after injection. RT-PCR and ELISA were used to determine the TNF α mRNA and protein expression; HE staining observation of tumor necrosis. Results: The model of SGC-7901 gastric cancer xenografts in nude mice was successfully established. The tumor volume of nude mice in three groups was (0.51 ± 0.27), (0.64 ± 0.36) and (1.21 ± 0.80) cm3, respectively. The tumor volume of UCBMSC-TNF-α group was the lowest (F = 3.88, P <0.05) The results of RT-PCR showed that the expression of TNF-αmRNA in UCBMSC-TNF-α group was (1.92 ± 0.12), (1.21 ± 0.26) and (0.81 ± 0.22) (F = 54.82, P <0.01). The results of ELISA showed that the expression of tumor necrosis factor-α in three tumor-bearing groups was 148.29 ± 3.76, (78.22 ± 6.24), (42.80 ± 3.02) pg / ml, the expression of MSC-TNF-α was the highest (F = 694.54, P <0.01). The pathological sections of HE staining showed that the tumor necrosis area was the largest in UCBMSC-TNF-α treated group. Conclusion: TNF-α transgenic UCBMSC can inhibit gastric cancer growth by paracrine TNF-α.