tumstatin基因修饰巨核细胞联合化疗抑制肺腺癌小鼠皮下移植瘤生长

Tumstatin transgenic megakaryocyte combined with chemotherapy inhibits angiogenesis in subcutaneous lung adenocarcinoma cancer xenografts in mice

目的探讨肿瘤抑素(tumstatin)转基因巨核细胞联合化疗对肺腺癌小鼠生存率的影响和皮下移植瘤生长的抑制作用。方法制备tumstatin转基因巨核细胞,通过小鼠背部接种肺腺癌A549细胞,建立小鼠肺腺癌移植瘤模型。小鼠皮下瘤长径达到5 mm后随机分为4组:生理盐水对照组(A组,对照组),吉西他滨(GEM)化疗组(B组,单纯化疗组)、GEM联合巨核细胞组(C组)、GEM联合tumstatin转基因巨核细胞组(D组);其中B、C、D三组为治疗组。对各组小鼠分别进行给药24 d,每3 d测1次肿瘤体积。在治疗第15天眼球取血,通过血液分析仪检测血细胞值;第24天剥离瘤体,测量大小及质量,免疫组织化学法检测肿瘤内微血管密度(MVD),HE染色检测肿瘤组织坏死情况,治疗期间记录小鼠生存情况。结果各治疗组肿瘤体积生长曲线显著慢于生理盐水组,其中D组瘤体生长最慢且该组生存率较单纯化疗组(B组)得到明显提高;与其他三组相比,D组小鼠皮下瘤MVD受到显著抑制并引起肿瘤组织重度坏死,且相应小鼠体内保持较正常的血细胞水平。结论 tumstatin转基因巨核细胞联合化疗较单独化疗显著抑制肿瘤新生血管生长并可提升肺腺癌小鼠的生存率。

Objective To investigate the effect of tumstatin transgenic megakaryocytic cells combined with chemotherapy on the survival rate of lung adenocarcinoma mice and the inhibitory effect on the growth of subcutaneous xenografts. Methods Preparation of tumstatin transgenic megakaryocytes; an ectopic mouse model of lung adenocarcinoma was established by subcutaneously inoculating lung adenocarcinoma A549 cells into the back of mice. The mice were then randomly divided into 4 groups： normal saline control group was group A,gemcitabine chemotherapy group was group B,gemcitabine combined with megakaryocytes group was group C,gemcitabine combined with tumstatin transgenic megakaryocytes group was group D,and the latter three groups were the treatment group. The mice in each group were administered for 24 days,and the tumor volume was measured every 3 days. Blood samples were taken from the 15 th day,blood cell values were measured by blood cell analyzer. On day 24,the tumor was dissected and the size and quality were measured. The microvessel density（ MVD） was tested by immunohistochemistry. Tumor necrosis was detected by hematoxylin and eosin staining. Survival of mice was recorded during treatment. Results The tumor volume growth curve of the treatment group was significantly slower than that of the normal saline group. The survival rate of the tumstatin transgenic megakaryocytic cells was the slowest. The survival rate was significantly improved compared with the chemotherapy group（ group B）. Compared with the normal saline group and other treatment groups,group D significantly inhibited tumor MVD and caused severe necrosis of tumor tissue,and the corresponding mice maintained a normal blood cell level. Conclusion The combination of tumstatin monocyte megakaryocytosis can enhance the survival rate of lung adenocarcinoma mice and inhibit the MVD of lung adenocarcinoma significantly.