目 的 ：体 内 外 实 验 探 讨 木 鳖 子 单 体 化 合 物 对 羟 基 桂 皮 醛 [Momordica cochinchinensis(Lour.)Spreng. p[1]hydroxycinnamaldehyde，CMSP]对小鼠黑色素瘤移植瘤生长和转移的影响及其作用机制。方法：建立荷瘤小鼠动物模型，并将 18只C57BL/6小鼠随机分成3组（每组6只）：对照组（腹腔注射0.1 ml生理盐水）、CMSP治疗组（分别腹腔注射0.1 ml 1、2 mg/ml CMSP），给药的第5天开始，每次给药前用卡尺分别测量和计算小鼠移植瘤的体积，实验结束后称量移植瘤的质量；H-E染色后光 镜观察肝组织的病理学变化；免疫组织化学SP法观察移植瘤组织E-cadherin和vimentin蛋白的表达。采用细胞划痕和Transwell 实验分别检测CMSP实验组（10、20 μg/ml）黑色素瘤B16细胞24、48 h的迁移能力，qPCR法检测CMSP处理24 h后B16细胞EMT 相关mRNA表达，WB法检测CMSP处理B16细胞48 h后β-catenin、p-β-catenin（Ser675）、vimentin和E-cadherin蛋白的表达水平。 结果：CMSP治疗组小鼠移植瘤平均体积和肿瘤质量明显降低（均P P<0.05）, CMSP（2 mg/kg）处理组小鼠的肝组织内未发现明显转移灶。CMSP治疗组（1、2 mg/kg）移植瘤 组织中E-cadherin蛋白表达水平明显高于对照组（均P<0.05），而vimentin蛋白表达显著低于对照组（均P<0.01）。体外实验中， CMSP实验组（10、20 μg/ml）B16细胞24、48 h后划痕愈合率较对照组均明显降低（均P<0.05）。20 μg/ml CMSP处理B16细胞24、 48 h后穿过Transwell小室的细胞数较对照组则显著下降（均P<0.01）。CMSP（10、20 μg/ml）处理B16细胞后β-catenin mRNA表 达水平较对照组明显降低（均P<0.01）E-cadherin mRNA表达水平则明显升高（均P<0.05）, 而 vimentin mRNA表达水平在10 μg/ml 处理组与对照组相比差异无统计学意义（P>0.05），20 μg/ml处理组则明显降低（P<0.01）。与对照组相比，CMSP实验 组（10 、20 μ g/ml）处 理 B16 细 胞后β-catenin、p-β-catenin和vimentin蛋白表达均显著降低（均P<0.01），而E-cadherin蛋白表达 则明显升高（均P<0.01）。结论：CMSP能够抑制小鼠黑色素瘤移植瘤的生长和转移，其作用机制可能与抑制wnt/β-catenin通路 的活性相关。

Objective: To investigate the effect and mechanism of p-hydroxycinnamaldehyde (CMSP) momomer compounds extract from the Cochinchina momordica seeds on the growth and metastasis of melanoma transplanted tumors in mice with in vivo and in vitro experiments. Methods: A tumor-bearing mouse model was established, and 18 C57BL/6 mice were randomly divided into 3 groups (6 mice in each group): a control group (intraperitoneal injection of 0.1 ml normal saline) and two CMSP treatment groups (intraperitoneal injection of 1 and 2 mg/ml CMSP of 0.1 ml). From the 5th day of drug administration, the transplanted tumor in mice was measured with calipers and the volume was calculated before each administration. At the end of the experiment, the mass of transplanted tumor was weighed. The pathological changes of liver tissues were observed under light microscope after H-E staining. The expressions of E-cadherin and vimentin in the transplanted tumor tissues were detected by SP immunohistochemical method. The metastatic ability of melanoma B16 cells in the CMSP group (10, 20 μg/ml) was detected by cell scratch test and Transwell test after treatment for 24 and 48 h. The expressions of EMT-related mRNAs in B16 cells after treatment with CMSP for 24 h were detected by qPCR. The protein expression levels of β-catenin, p-β-catenin (Ser675), vimentin and E-cadherin in B16 cells treated with CMSP for 48 h were detected by Western blotting(WB). Results: The mean tumor volume and tumor mass of the mice in CMSP treatment group were significantly decreased (all P<0.05). The number of liver metastases in the control group was significantly higher than that in the CMSP (1 and 2 mg/ kg) groups (all P<0.05); moreover, no obvious metastases were found in the liver tissues of the 2 mg/kg CMSP treatment group. The expression level of E-cadherin protein in CMSP group (1 and 2 mg/kg) was significantly higher than that in the control group (all P<0.05), while the expression level of vimentin protein was significantly lower than that in the control group (all P<0.01). For the in vitro experiments, the scratch healing rate of B16 cells in CMSP groups (10, 20 μg/ml) after treatment for 24 and 48 h was significantly lower than that in control group (all P<0.05). The number of B16 cells passing through Transwell compartment in the 20 μg/ml CMSP treatment group was significantly decreased as compared with the control group (all P<0.01). The mRNA expression level of β-catenin in B16 cells treated with CMSP (10, 20 μg/ ml) was significantly lower than that in control group (all P<0.01), while the mRNA expression level of E-cadherin was significantly increased (all P<0.05); However, the mRNA expression level of vimentin in the 10 μg/ml treatment group was not significantly different from that in the control group (P>0.05), but it was significantly decreased in the 20 μg/ml treatment group (P<0.01). Compared with the control group, the protein expressions of β-catenin, p-β-catenin and vimentin in B16 cells of CMSP groups (10 and 20 μg/ml) were significantly decreased (all P<0.01), while the protein expression of E-cadherin was significantly increased (all P<0.01). Conclusion: CMSP can inhibit the growth and metastasis of melanoma transplanted tumors in mice, and its mechanism may be related to the activity of wnt/β-catenin pathway.

国家自然科学基金资助项目（No.81973520）