从OPG/RANKL/RANK系统阐述补骨脂-蛇床子抑制乳腺癌骨转移的机制

作者	单位
杨顺芳	上海交通大学附属胸科医院核医学科
吴春宇	上海中医药大学龙华医院中医外科
宋晓耘	上海中医药大学龙华医院中医外科
刘胜	上海中医药大学龙华医院中医外科
程旭锋	上海中医药大学龙华医院中医外科

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中文摘要:

目的从OPG/RANKL/RANK系统探讨不同比例补骨脂-蛇床子抑制乳腺癌骨转移的机制,发掘补骨脂-蛇床子抑制乳腺癌骨转移最佳配伍比例。方法 MDA-MB-231BO注射裸鼠左心室制造乳腺癌骨转移模型,随机分为补骨脂:蛇床子4∶0组、3∶1组、1∶1组、1∶3组、0∶4组、唑来磷酸组、生理盐水组,实验组在造模后1周给予中药煎剂灌胃干预,阳性对照组造模后1周腹腔注射唑来磷酸,阴性对照组造模后1周灌胃生理盐水。用药6周后,通过放射性核素骨显像和X-ray摄片找到转移病灶,然后取病变组织进行RT-PCR及Western-blot检测护骨素(osteoprotegerin,OPG)、核因子κB的受体激活剂配体(receptor activator of NF-κB ligand,RANKL)、白介素8(interleukin-8,IL-8)、甲状旁腺激素相关蛋白(parathyroid hormone-relatedpeptide,PTHrP)、巨噬细胞集落因子(macrophage colony stimulating factor,MCSF)的基因及蛋白表达量。结果蛇床子、补骨脂及补骨脂-蛇床子不同配伍比例均对抑制骨转移基因有一定的作用,OPG的表达量增高(蛇床子∶补骨脂1∶1组mRNA的表达量(60.343±6.274),IL-8、PTHrP、MCSF、RANKL的表达量均下降(蛇床子∶补骨脂1∶1组IL-8、PTHrP、MCSF、RANKL mRNA的表达量分别是218.010±12.802,232.399±14.354,319.831±5.322,195.701±4.862。中药蛇床子∶补骨脂1∶1组对相关抑制骨转移基因的mRNA表达量作用效果最为明显,与生理盐水组比较差异有统计学意义(P<0.01)。同时,蛇床子、补骨脂及蛇床子∶补骨脂不同配伍比例均能够上调骨转移灶中OPG、下调RANKL、IL-8、PTHrP、MCSF的蛋白表达。其中,补骨脂∶蛇床子1∶1组的效果最为明显,与生理盐水组比较差异有统计学意义。结论补骨脂-蛇床子通过激活OPG/RANKL/RANK通路,抑制乳腺癌骨转移,其中1∶1组效果最为明显,为最佳配伍比例。

英文摘要:

Objective To find the optimal proportion of Composite Fructus Psoraleae and Fructus Cnidii (CFPC) for inhibiting the bone metastasis of breast cancer by way of exploring its acting mechanism viewing from OPG/RANKL/RANK system. Methods The human bone metastasis of breast cancer model was established by injecting tumor cells of MDA-MB-231BO cell line into the left cardiac ventricle of nude mice. The modeled mice were randomly divided into seven groups:the blank group administered with normal saline by gastrogavage,the positive control group with zoledronic acid via peritoneal injection,and the 5 tested group with CFPC in different proportions of Fructus Psoraleae and Fructus Cnidii,i.e. (A,4:0; B,3:1; C,1:1; D,1:3,and E 0:4),given by gastric infusion. The treatment started from 1 week after modeling and lasted for six weeks. By the end of the experiment,the metastatic foci in bone were imaged by radionuclide tracing method and X-ray photograph,and separated for detecting gene and protein expressions of osteoprotegerin (OPG),receptor activator of nuclear factor-κB ligand (RANKL),interleukin-8 (IL-8),parathyroid hormone-related protein (PTHrP),macrophage colony stimulating factor (MCSF) by Real-time PCR and Western blot respectively. Results Inhibition of bone metastasis gene was displayed to some extent in all the tested groups treated with CFPC,showing an increased level of OPG mRNA expression (It was 60.343±6.274 in the tested group C),and decreased mRNA expressions of IL-8,PTHrP,MCSF,RANKL (218.010±12.802,232.399±14.354,319.831±5.322,and 195.701±4.862,respectively in the tested group C). The optimal effect was shown in the tested group C,showing significant difference to that in the blank group (P<0.01). Meanwhile,the OPG in the bone metastatic foci could be up-regulated and protein expressions of RANKL/IL-8/PTHrP/MCSF down-regulated in all the tested groups. The optimal effect was shown in the tested group C,with significant difference from those of the normal saline group. Conclusion CFPC could inhibit the bone metastasis of breast cancer through activating OPG/RANKL/RANK pathway. Among different proportions of Fructus Psoraleae and Fructus Cnidii,1:1 was the best one.

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