反应停对小鼠肝癌细胞H22移植瘤生长的影响

Effect of Thalidomide on Tumor Growth in Mouse Hepatoma H22 Model

背景与目的:反应停因具有抗血管生成作用而可能对某些肿瘤有治疗作用。本研究旨在通过观察反应停对小鼠H22移植瘤生长的影响,探讨反应停的作用机制。方法:BALB/c小鼠皮下接种H22细胞,分别从接种当天(第一给药组)和第4天(第二给药组)开始每天腹腔注射反应停50mg/kg,每天测肿瘤直径,第12天处死动物,称瘤重。对移植瘤组织,用抗CD31单抗做免疫组化染色,测定微血管密度;流式细胞术分析CD31表达以及凋亡率;逆转录聚合酶链反应(reversetranscription-ploymerasechainreaction,RT-PCR)检测血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)mRNA的表达。结果:第一给药组的瘤重为(2.27±0.33)g,小于对照组的(2.70±0.61)g,但差异没有统计学意义(P>0.05),抑瘤率为15.93%;第二给药组的瘤重为(3.32±0.47)g,小于对照组的(3.42±0.60)g,差异也无统计学意义(P>0.05),抑瘤率为2.92%。第一给药组移植瘤组织中微血管计数和CD31表达分别为(48.4±12.90)和(5.59±0.75),与对照组(81.2±26.91)和(7.04±0.50)比较均显著降低(P<0.05);第二给药组微血管计数和CD31表达分别为(46.4±9.71)和(7.29±0.48),与对照组的(74.0±32.69)和(6.80±0.68)相比,差异无统计学意义(P>0.05)。两给药组移植瘤组织细胞凋亡指数分别为(17.

BACKGROUND &OBJECTIVE:Thalidomide may have curative effect on t um or because of its function of anti-angiogenesis. The aim of this research was t o observe the effect of thalidomide on tumor homograft growth in mouse H22 model and to investigate the mechanisms involved and its curative possibility to hepa toma.METHODS:BALB/c mice were inoculated subcutane-ously with H22 cells. The fi rst treatment group was injected intraperitoneal everyday with thalidomide 50 mg /kg from the day of inoculation and the second treatment group was injected from the fourth day of inoculation. The diameters of the tumors were measured everyd ay. On the twelfth day,the mice were sacrificed and the tumors were weighed. The microvessel densities of the tumors were measured by immunohistochemical staini ng with anti-CD31 monoclonal antibody; CD31 expression, apoptosis, and prolifer ation were analyzed by flow cytometry. Expression of vascular endothelial growth factor (VEGF) mRNA was examined by reverse transcription-polymerase chain reac tion (RT-PCR). RESULTS:The tumor weight of the first treatment group (2.27±0.3 3 g) was decreased compared with that of control group(2.70±0.61 g) (P >0 05); the ratio of inhibition was 15.93%. The tumor weight of the second treatment gr oup (3.32±0.47 g) was decreased compared with that of control group (3.42±0.60 g) (P >0.05);the ratio of inhibition was 2.92%.The microvessel count (48.40±1 2.90) and CD31 expression (5.59±0.75) of the first treatment group were signifi cantly decreased (P< 0.05) in comparison with those of control groups (81.20±26 .91,7.04±0.50)(P< 0.05). The microvessel count (46.4±9.71) and CD31 expression (7.29±0.48) of the second treatment group were not statistically significant ( P >0.05) compared with those of control groups (74.0±32.69,6.80±0.68)(P >0.05) . The apoptosis indices of the two treatment groups (17.20±7.80,15.33±4.10) we re significantly increased, compared with control groups (4.37±1.98,4.87±1.91) (P< 0.05), while there was no significant difference of VEGF mRNA expression bet ween the two treatment groups and control groups (P >0.05). The relative quantit ies of VEGF mRNA of the two treatment groups were 0.50±0.13 and 0.51±0.06 (con trol groups:0.48±0.11 and 0.64±0.11)(P >0.05), respectively. CONCLUSION: Thali domide can significantly induce apoptosis and inhibit angiogenesis in mouse H22 model when it is used at the beginning of carcinogenesis, but it has no obvious anti-angiogenic efficacy on the grown tumor. Thalidomide cannot inhibit VEGF mR NA expression of grafted H22 tumor in mouse.