郭文韬 张燕

[摘要] 目的 探讨片仔癀对裸鼠人骨肉瘤肿瘤干细胞移植瘤的抑制作用及其对ABCB1、ABCG2蛋白表达的影响。 方法 48只裸鼠皮下接种肿瘤干细胞建立人骨肉瘤肿瘤干细胞移植瘤模型。依照随机数字表法将其分为对照组、阿霉素组、片仔癀组、片仔癀与阿霉素联合组，每组各12只，分别予以相应的干预，4周后处死裸鼠，测量瘤体体积及称取其重量，计算肿瘤体积及重量抑制率；免疫组织组化学（SP法）及Western blot检测瘤体内ABCB1、ABCG2的表达。 结果 与对照组比较，阿霉素组、片仔癀组、联合组瘤体体积及重量明显降低（P < 0.05），与片仔癀组比较，联合组生长受到明显抑制（P < 0.01）；与对照组比较，片仔癀组、联合组可明显降低ABCB1、ABCG2的表达（P < 0.05），且联合组较片仔癀组明显下调（P < 0.05）。 结论 片仔癀可明显抑制人骨肉瘤肿瘤干细胞裸鼠移植瘤的生长，且能够增强阿霉素的作用，其作用机制与抑制膜转运蛋白ABCB1、ABCG2的表达相关。

[关键词] 片仔癀；骨肉瘤；干细胞移植瘤；影响

[中图分类号] R738.1 [文献标识码] A [文章编号] 1673-7210（2018）04（a）-0004-05

[Abstract] Objective To explore the inhibitory effect of Pien Tze Huang on the growth of human osteosarcoma stem cells transplantation tumor of nude mice and its effect on ABCB1 and ABCG2 protein expression. Methods The model of human osteosarcoma tumor stem cell transplantation tumor was established by subcutaneous tumor stem cells in nude mice. According to the random number table method ， 48 nude mice were divided into control group， adriamycin group， Pien Tze Huang group， Pien Tze Huang and adriamycin combined group， with 12 mice in each group， The nude mice were executed after four weeks of intervention. The volume and weight of the tumor were measured and the tumor volume and weight inhibition rate were calculated. The expression of ABCB1 and ABCG2 in the tumor was detected by immunohistochemistry （SP） and Western blot. Results Compared with the control group， the volume and weight of tumor were significantly reduced in theadriamycin group， Pien Tze Huang group and combined group （P < 0.05）. Compared with Pien Tze Huang group， the combined group growth was significantly inhibited. （P < 0.01）. SP and Western blot results showed that the expressions of ABCB1 and ABCG2 in the Pien Tze Huang group and combined group were significantly down-regulated than that in the control group （P < 0.05）， and combined group was significantly lower than that in the Pien Tze Huang group （P < 0.05）. Conclusion Pien Tze Huang can significantly inhibit the growth of tumor stem cells in human osteosarcoma tumor stem cells， and it can enhance the effect of adriamycin. Which may be related to inhibit the expression of membrane transporter ABCB1 and ABCG2.

[Key words] Pien Tze Huang； Osteosarcoma； Stem cell transplantation tumor； Effect

骨肉瘤好發于青少年，恶性程度高，早期即可发生转移，尽管随着医疗技术的提高，目前患者5年生存率可达到70%，但仍有30%患者因对化疗药物的耐药而导致治疗失败[1]。前期大量的研究表明多种机制与骨肉瘤化疗耐药的产生有关[2-3]，包括ABC转运蛋白的过表达，细胞凋亡调控异常，细胞内解毒系统紊乱及肿瘤干细胞的存在等[4-5]。研究发现肿瘤干细胞能通过上调ABC转运蛋白的表达等作用，使其对化疗药物具有耐药性，成为肿瘤耐药的重要原因[6]。我们前期实验发现骨肉瘤干细胞中高表达ABCB1、ABCG2蛋白，片仔癀可明显抑制耐药骨肉瘤干细胞的增殖，抑制ABCB1、ABCG2蛋白表达，逆转其耐药[7]。本实验通过观察片仔癀对裸鼠人骨肉瘤肿瘤干细胞移植瘤及其ABCB1、ABCG2的表达影响，进一步探讨片仔癀对骨肉瘤耐药性的作用。……