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目的:探讨黄芪解毒汤体外干预对人乳腺癌细胞(MCF-7)的SHH、PTCH1、SMO及GLI1基因和特异性周期蛋白-D1(CyclinD1)表达的影响。方法:制备含药血清,分为黄芪解毒汤血清组、阿霉素血清组、参一胶囊血清组、生理盐水血清组。常规培养细胞。分别以20%浓度不同含药血清干预细胞。实时荧光定量PCR检测SHH、PTCH1、SMO及GLI1基因表达的影响,免疫荧光检测CyclinD1表达的影响。结果:与生理盐水血清组比较,含药血清组SHH、PTCH1、SMO及GLI1基因表达均明显降低(P<0.01);阿霉素血清组抑制率最强(P<0.01);黄芪解毒汤血清组与参一胶囊血清组比较,差异无统计学意义(P>0.05);含药血清组中CyclinD1蛋白的表达低于生理盐水血清组,差异有统计学意义(P<0.01);阿霉素血清组CyclinD1蛋白表达抑制率最强(P<0.01),黄芪解毒汤血清组与参一胶囊血清组比较,差异无统计学意义(P>0.05)。结论:黄芪解毒汤可诱导乳腺癌细胞MCF-7凋亡,显著降低SHH、PTCH1、SMO及GLI1基因和CyclinD1蛋白表达。
Objective: To investigate the effects of Huangqi Jiedu Decoction on expression of SHH, PTCH1, SMO, GLI1 and CyclinD1 in human breast cancer cells (MCF-7) in vitro. Methods: Preparation of serum containing astragalus detoxification soup serum group, doxorubicin serum group, a capsule of serum group, saline group. Cells are cultured routinely. Respectively with different concentrations of 20% serum intervention cells. The effects of SHH, PTCH1, SMO and GLI1 gene expression were detected by real-time fluorescence quantitative PCR, and the effect of CyclinD1 expression was detected by immunofluorescence. Results: Compared with the saline group, the expression of SHH, PTCH1, SMO and GLI1 in the serum-containing group were significantly decreased (P <0.01), and the inhibition rate in the doxorubicin serum group was the strongest (P <0.01) (P> 0.05). The expression of CyclinD1 protein in the serum-containing group was lower than that in the saline group (P <0.01), but there was no significant difference between the two groups The inhibition rate of CyclinD1 protein in serum group was the strongest (P <0.01). There was no significant difference between Huangqi Jiedu Decoction serum group and Shenyi Capsule serum group (P> 0.05). Conclusion: Astragalus and Jiedu Decoction can induce the apoptosis of breast cancer cell MCF-7 and significantly reduce the expression of SHH, PTCH1, SMO, GLI1 and CyclinD1.