从CXCL12/CXCR4生物学轴探讨乳移平配伍引经药抗乳腺癌肺转移机理体外研究

Effect and Mechanism of Ruyiping Formula plus Platycodon Grandiflorum in Treating with Breast Cancer Lung Metastasis Cell Lines from CXCL12/CXCR4 Biological Axis in vitro

目的:探讨"乳移平"配伍肺经引经药"桔梗"对人乳腺癌肺高转移细胞株MDA-MB-231HM的增殖、趋化、迁移能力的影响及作用机制。方法:乳移平组、乳移平配伍桔梗组、乳移平配伍川牛膝组、阿霉素组和对照组SD大鼠含药血清的制备,MTT法测不同组含药血清对细胞增殖能力的影响,Transwell小室测细胞趋化活性的影响,实时定量PCR法测细胞CXCL12、CXCR4、PI3K的表达。结果:与乳移平单方比,乳移平配伍桔梗后可进一步抑制MDA-MB-231HM的增殖能力(P<0.01)和趋化活性(P<0.01)。实时定量PCR结果提示,与乳移平单方比,乳移平配伍桔梗后能进一步下调MDA-MB-231HM细胞株CXCL12、CXCR4基因(P<0.05)。结论:乳移平配伍桔梗通过进一步下调CXCL12/CXCR4生物学轴及其介导的PI3K信号转导通路,发挥进一步的抑制乳腺癌肺高转移细胞株增殖能力和趋化活性的作用。

Objective： To observe the effect and mechanism of Ruyiping Formula plus platycodon grandiflorum,an herb guiding others to lung,in the treatment of lung metastasis of breast cancer MDA-MB-231 HM cell lines in vitro. Methods： We prepared the SD mice serum with different drugs administrated,including Ruyiping（ basic fomula）,Ruyiping plus platycodon grandiflorum,Ruyiping plus radix cyathulae,normal saline control and doxorubicin groups. The effects of different drug serums on MDA-MB-231 HM cell lines were studied by MTT assay. The changes of chemotactic activity on MDA-MB-231 HM cell lines were detected by transwell chamber method. The mRNA expressions of CXCL12,CXCR4 and PI3 K genes of MDA-MB-231 HM cell lines which were intervened by different herb serums were observed with real-time quantitative PCR. Results： Compared with Ruyiping,Ruyiping plus platycodon grandiflorum could further inhibit the reproductive activity（ P〈0. 01） and chemotactic activity（ P〈0. 01） on MDA-MB-231 HM cell lines. Real-time quantitative PCR revealed that Ruyiping plus platycodon grandiflorum group could further down-regulate the CXCL12 and CXCR4 expressions（ P〈0. 05） of MDA-MB-231 HM cells. Conclusions： Ruyiping plus platycodon grandiflorum could further down-regulate the CXCL12 / CXCR4 axis and be mediated by PI3 K downstream signaling pathway in order to intervene the reproductive activity and chemotactic activity on breast cancer lung metastasis cell lines.