恶性黑色素瘤是最难治的肿瘤之一。槐花金银酒出自《医学启蒙》是以清热解毒为法治疗皮肤疾病的古方。我们的初步研究表明：槐花金银酒能抑制荷瘤小鼠黑色素瘤生长，升高let-7家族的抑癌miRNAs let-7a及let-7f并降低胰岛素样生长因子1受体（IGF1R，let-7a及let-7f的靶基因）的表达及PI3K/Akt（IGF1R下游分子）通路的激活。基于此，我们提出以下工作假说：槐花金银酒通过上调let-7a/f抑制靶基因IGF1R 及其下游的信号通路而抑制黑色素瘤生长。本研究拟采用反义RNA及过表达技术，分别构建抑制let-7a/f和上调IGFR1及激活PI3K的黑色素瘤稳定细胞系，并在体内、外对比调控上述分子后槐花金银酒抑制黑色素瘤生长的差异，验证let-7a/f、IGF1R以及PI3K/Akt在槐花金银酒抗黑色素瘤中的作用。结果将为该方抗黑色素瘤提供分子机制。

Malignant melanoma is one of the most treatment-resistant cancers. Huai Hua Jin Yin Jiu (HHJYJ) is a traditional formula which is first recorded in Yi Xue Qi Meng. It is used to treat skin diseases following the TCM therapeutic principle of clearing away heat and toxic materials. Our preliminary investigations demonstrated that HHJYJ could significantly inhibit tumor growth in B16F10 melanoma-baring mice. HHJYJ treatment could also upregulate let-7a and let-7f (tumor suppressing miRNAs), lower the expression level of insulin-like growth factor 1 receptor (IGF1R, a let-7a/f target) and activate the PI3K/Akt (a downstream pathway of IGF1R signaling) pathway. We hypothesize that the let-7/IGF1R pathway is involved in the anti-melanoma action of HHJYJ. To address this hypothesis, we propose to use antisence and overexpression approaches to establish stable melanoma cell lines and then determine whether knockdown let-7a/f or overexpress IGF1R or a constitutively active form of PI3K can weaken/abolish the anti-melanoma action of HHJYJ in vitro and in vivo. It is expected to establish the role of the let-7/IGF1R pathway in the anti-melanoma action of HHJYJ, providing molecular mechanisms of action for this formula.