沙利度胺联合表阿霉素对小鼠H22肝癌移植瘤的生长抑制研究

Mechanism and inhibition of thalidomide combined with epirubicine on the growth of transplanted H22 hepatocellular carcinoma in mice

目的研究沙利度胺联合表阿霉素对小鼠H22肝癌移植瘤的生长抑制作用。方法用H22肝癌腹水瘤细胞构建荷瘤肝癌小鼠模型。40只模型小鼠随机分为4组,每组10只:生理盐水组(0.3μL·d^(-1)),表阿霉素组(50 mg·m^(-2)),沙利度胺组(200 mg·kg^(-1)·d^(-1)),沙利度胺联合表阿霉素组(沙利度胺200 mg·kg^(-1)·d^(-1)+表阿霉素50 mg·m^(-2)),表阿霉素灌胃1次,其余连续腹腔注射10 d,每隔7 d测量移植瘤长短径,并于第28天处死小鼠,用反转录聚合酶连锁反应法检测各组移植瘤组织B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)及血管内皮生长因子(VEGF)mRNA含量,免疫印迹法检测各组移植瘤组织Bax和Bcl-2蛋白含量,免疫组化检测各组移植瘤组织VEGF蛋白表达。结果给药21 d后,表阿霉素组(0.67±0.01)、沙利度胺组(0.62±0.02)、联合组(0.43±0.01)中移植瘤体积较生理盐水组(0.90±0.01)明显减小(P<0.05),联合用药组移植瘤体积较单独给药组明显减小(P<0.05)。给药27 d后,表阿霉素、沙利度胺及联合组移植瘤组织中Bcl-2及VEGF mRNA及蛋白表达量较生理盐水组明显降低(P<0.05),Bax mRNA及蛋白表达量较较生理盐水组明显提高(P<0.05),同时联合用药组与单独给药组比较差异有统计学意义(P<0.05)。结论沙利度胺及表阿霉素具有明显的抑制肿瘤作用,机制可能与上调VEGF、Bcl-2表达和下调Bax表达有关,并且具有协同作用。

Obejective To explore mechanism and inhibition of thalidomide（ Tha） combined with epirubicine（ Epi） on the growth of transplanted H22 hepatocellular carcinoma in mice. Methods Forty male Kunming mice were transplanted by right armpit injection of 200 μL（ 2. 0 ×10^6cells） H22 tumor cells. Then these mice were randomly divided into four groups,ten mice in each group. Saline group,Tha group（ oral 200mg·kg^-1·d^-1for 10 consecutive days）; Epi group（ intra-peritoneall injection,50 mg·m^-2,once）,Tha combined with Epi group（ oral Tha200 mg·kg^-1·d^-1,for 10 consecutive days; Epi intraperitoneall injection,50 mg· m^-2,once）. The tumor diameter and short diameter was measured every 7 d,and twenty- eighth days were sacrificed by dislocation,tumor tissue was obtained. The expression of B- cell lymphoma- 2（ Bcl- 2）,Bcl- 2 associated X protein（ Bax）,and vascular endothelial growth factor（ VEGF） mRNA was measured by RT- PCR. The expression of Bax,Bcl- 2protein was measured by Western blot. The expression of VEGF protein was measured by immunohistochemical.Results After 21 days,compared with saline group（ 0. 90 ± 0. 01）,Epi group（ 0. 67 ± 0. 01）,Tha group（ 0. 62 ± 0. 02）and combined group（ 0. 43 ± 0. 01） transplanted tumor volume was decreased（ P〈0. 05）. Compared with Epi group and Tha group,combined group transplanted tumor volume was decreased（ P〈0. 05）. Compared with saline group,the expression of Bcl- 2 and VEGF mRNA and protein was down- regulated（ P〈0. 05）,the expression of Bax mRNA and protein was up- regulated in Epi group,Tha group and combined group（ all P〈0. 05）. The expression of Bax,Bcl- 2 and VEGF mRNA and protein had significant difference in combined group compared with Tha and Epi group（ P〈0. 05）.Conclusion Tha and Epi could inhibit the growth of transplanted H22 hepatocellular carcinoma,related to lower VEGF and Bcl- 2 expression as well as higher Bax expression,and Tha combined with Epi might have synergistic effect.