载药纳米炭的淋巴化疗对小鼠结肠癌转移模型肿瘤细胞凋亡的影响

Research of lymphatic chemotherapy of drug loaded carbon nanoparticles to colonic carcinoma metastasis model in mice

目的应用载药纳米炭对小鼠结肠癌转移模型进行淋巴化疗,探讨其对肿瘤细胞凋亡的影响。方法采用小鼠爪垫接种小鼠结肠低分化腺癌Colon 26细胞悬液的方法,建立小鼠结肠癌淋巴结转移模型。将成功制备的40只荷瘤小鼠随机分为4组,每组10只,爪垫肿瘤处皮下分别注射5%葡萄糖溶液(A组)、纳米炭-氟尿嘧啶混合液(B组)、纳米炭-奥沙利铂混合液(C组),纳米炭-氟尿嘧啶混和液及纳米炭-奥沙利铂混合液联合用药(D组)。观察小鼠爪垫注射部位局部一般情况,给药48小时后处死小鼠,取爪垫肿瘤组织制成单细胞悬液,采用流式细胞分析法检测癌细胞凋亡率;同时取腘窝淋巴结,应用原位末端脱氧核苷转移酶标记法(TUNEL法)检测淋巴结转移癌细胞凋亡指数。结果注射药物后各组小鼠无死亡,爪垫注药部位无溃疡及坏死;B、C组和D组小鼠腘窝淋巴结黑染。B、C组和D组爪垫癌细胞凋亡率和腘窝淋巴结转移癌细胞凋亡指数与A组比较有显著性差异(P<0.05),D组与B、C组比较差异显著(P<0.05),B组与C组比较差异不显著(p﹥0.05)。结论载药纳米炭能够靶向性进入淋巴结内,并在局部滞留,能诱导种植肿瘤及淋巴结转移癌细胞凋亡,纳米炭载氟尿嘧啶与纳米炭载奥沙利铂联合用药有协同增效诱导肿瘤细胞凋亡的作用。

Objective To investigate the effect of the lymphatic chemotherapy to colonic carcinoma metastasis model in mice by drug loaded carbon nanoparticles.Methods The colonic carcinoma metastasis model in mice were established by injection of colon26 cell suspension and divided into 4 groups randomly.5% glucose（A group）,injection of fluorouracil and carbon nanoparticles（B group）,injection of oxaliplatin and carbon nanoparticles（C group）,mixing of injection of fluorouracil and carbon nanoparticles and injection of oxaliplatin and carbon nanoparticles（D group） were injected into paw pads with tumor,respectively.Observe the injection site of pad.Mice were executed after 48 hours.The rate of apoptosis of cancer cells was analyzed by Flow cytometry.Apoptosis index of metastasis was tested by TUNEL.Results There was no death and no skin erosion on paw pads in all groups.Popliteal fossa lymph node was stained black in group B,C and D.The rate of apoptosis in metastasis and apoptosis index in popliteal fossa lymph node in group B,C and D is higher than group A（P0.05）,Differences between group D and group B,C was significant（P0.05）,there was no significant difference in group B and C（P0.05）.Conclusion Drug loaded carbon nanoparticles can be imported and detained in lymph node.It can induce apoptosis of tumor and lymph node metastasis,The synergistic effect of anti-cancer can be promoted by the combination of drug loaded carbon nanoparticles with 5-fluorouracil and oxaliplatin.