摘要
目的证实脐血造血干/祖细胞移植重建SCID鼠的免疫功能对人肝癌有抑制及杀伤作用。方法首先进行人肝癌高转移细胞株HCCLM6细胞培养,然后将HCCLM6细胞株分别在脐血移植免疫功能重建组(A组)及未移植脐血造血干/祖细胞组(B组)建立SCID鼠皮下荷瘤模型。接种肿瘤后5周处死,分别进行以下检测:(1)比较A、B两组间肿瘤生长情况的差异;(2)常规病理学检查:比较A、B两组移植瘤状况,并观察有无肿瘤转移;(3)免疫组化检查:观察肿瘤组织中人源CD3+、CD8+细胞对肿瘤组织的杀伤力;(4)细胞毒试验:A、B两组动物脾脏细胞与HCCLM6细胞体外混合培养,采用MTT法观察对肿瘤细胞的杀伤作用。结果 (1)HCCLM6细胞株接种于A组、B组SCID小鼠皮下均可成瘤,成瘤率为100%,成瘤潜伏期分别为(9.83±1.47)d、(5.16±1.17)d,二组比较差异有统计学意义(P<0.01);接种后5周移植瘤体积分别为(539.22±94.08)mm3、(1 126.17±154.75)mm3,二组比较差异有统计学意义(P<0.01)。(2)病理学检查可见:A组肿瘤坏死明显,未见远处转移;B组坏死程度明显较轻,可见肺转移。(3)免疫组织化学检查:A组肿瘤组织中可见大量人源CD3+、CD8+细胞浸润,坏死组织周围更加明显;B组移植瘤内亦有肿瘤坏死,但未见人源淋巴细胞。(4)细胞毒试验表明:A组脾脏细胞对HCCLM6细胞的杀伤作用明显,在效靶比例为120∶1时,特异性杀伤率为(7.41±1.07)%;B组脾脏细胞对肝癌细胞的杀伤率为(3.97±0.27)%,二组比较差异有统计学意义(P<0.01)。结论HCCLM6是一个活力强、代谢活跃的肿瘤细胞亚群。人源脐血干细胞在SCID鼠体内重建免疫后可发挥对HCCLM6细胞的杀伤作用,对肝癌的生长有抑制作用。脐血干细胞移植有可能成为肝癌治疗的一种新方法,其应用前景值得进一步研究探讨。
Abstract Objective to prove ganulocyte-macrrophage colonyatimulating dry/progenitor cells transplantation reconstruction SCID mile immune function to human liver has inhibitory and kill role. Methods Human liver high transfer lines HCCLM6 cell was cultured at first, and then HCCLM6 lines respectively in umbilical cord blood transplantation immunity function reconstruction group （group A） and not transplant ganulocyte-macrrophage colonyatimulating dry/progenitor cell group （group B） were established SCID mice subcutaneous tumor-burdened model. Vaccination tumor hind 5 weeks ex- ecuted, and separately below detection. Compared with A and B between groups tumor growth situa- tion differences. Conventional pathology examination., compare A and B two groups of transplantation tumor condition, and observe whether tumor metastasis. Immunohistochemical check： observation of the tumor RenYuan CD3＋ , CD8＋ cells for tumor tissue damage. Cytotoxic test： A,B two groups of animals spleen cells and HCCLM6 cells in vitro mixed cultivating, using the MTT method observation of tumor cells kill role. Results HCCLM6 lines inoculation in group A, B group SCID mice subcutane- ous tumor, all can into into tumor rate for 100 % respectively, into tumor incubation period （9. 834±1.47） d, （5, 16±1.17） d, two groups of comparisons P〈0.01, After inoculation 5 weeks transplant tumors secreting TSH respectively （plus or minus 94.08） 539.22 ram3, （1126. 17±154.75） ram3, two groups of comparisons （P〈0. 01）, Were statistically significant. Pathology checks visible the group A tumor necrosis obvious, did not see the distant metastasis, Group B necrosis significantly lighter, visible pulmonary metastasis. Immunohistochemical check the group A tumor tissue which numerous RenYuan CD3＋ ,CD8＋ cells infiltration, necrotic tissue around more evident, and Group B transplant tumour inside also have tumor necrosis, but did not see RenYuan lymphocytes. Cytotoxic experiments indicated that A group of spleen cells to HCCLM6 cell damage, an obvious effect in effect target ratio for 120：1, specificity damage rate for （7.41± 1.07） %, group B splenic cell of liver canc- er ceil damage rate for （3. 97±0. 27） %, two groups of comparisons （P〈0. 01）. Conclusion HC-CLM6 is a strong vitality, metabolic activity of tumor cells subsets. RenYuan umbilical cord blood stem cells in SCID mice immune to rebuild after can give full play to HCCLM6 cell damage effect, of liver cancer growth inhibition. Umbilical cord blood stem cell transplantation may be liver cancer treat- ment of a new method, the application of this method is worthy of further research prospect.
出处
《贵州医药》
CAS
2011年第10期871-875,共5页
Guizhou Medical Journal
关键词
HCCLM6
脐血造血干/祖细胞移植
SCID小鼠
免疫重建
原发性肝癌
HCCLM6 Ganulocyte-macrrophage colonyatimutating dry/progenitor cell trans-plantation SCID mice Immune rebuilt Primary liver cancer
