熊果酸纳米脂质体微粒对小鼠黑色素瘤细胞生长的抑制作用及其机制

Effect of ursolic acid phospholipids nanopartides on the growth and the expression of MMP-9 of B16 melanoma ceils

目的探讨熊果酸纳米脂质体微粒（UA-PL-NP）对体外培养的黑色素瘤细胞生长的抑制作用及其机制。方法体外培养小鼠黑色素瘤细胞（B16），采用噻唑蓝（MTT）比色法观察熊果酸纳米脂质体微粒对其增殖活性的影响，Western blot法观察其对基质金属蛋白酶（MMP）-9表达的影响，同时采用流式细胞术检测细胞凋亡情况。实验中采用顺铂作为阳性对照。结果MTT检测熊果酸纳米脂质体微粒对B16细胞增殖有明显抑制作用，并呈浓度和时间依赖性，UA-PL-NP作用24hIC50值为7．68mg／L，顺铂作用24hIC卯值为9．33mg／L；流式检测UA-PL-NP浓度为7．68mg／L，作用24h后B16鼠黑色素瘤细胞的凋亡率为（53．20±7．13）％；浓度9．33mg／L顺铂作用24h的凋亡率为（36．10±5．85）％；阴性对照组24h凋亡率为（7．87±2．31）％；Western blot检测对照组MMP-9蛋白含量是UA-PL-NP组3．106倍，是顺铂组的2．022倍；顺铂组MMP-9蛋白含量是UA-PL-NP组的1．537倍。结论熊果酸纳米脂质体微粒能促进瘤细胞的凋亡从而抑制B16鼠黑色素瘤细胞的增殖，并降低MMP-9蛋白的表达，具有抑制肿瘤侵袭和转移的作用。

Objective To study the inhibitory effects and mechanisms of ursolic acid phospholipids nanoparticles on the growth and the expression of MMP-9 of B16 melanoma cells. Methods After B16 melanoma cells were treated with various concentrations of UA-PL-NP, inhibitory rate of the growth was measured by MTT assay. Apoptosis rate was analyzed by flow cytometry. The expression of MMP-9 protein was detected hy Western hlot. The positive group was treated with C-DDP. Results Proliferation of B16 melanoma cells in vitro was significantly was inhibited by ursolic acid phospholipids nanoparticles in a time-and dose-dependent manner. After B16 melanoma cells were treated with UA-PL-NP for 24 h, the IC50 value was 7.68 mg/L. The IC50 value of C-DDP was 9.33 mg/L for 24 h. After B16 melanoma cells were treated with 7.68 mg/L UA-PL-NP for 24 h, apoptosis rate was （53.20 ± 7.13）%. The apoptosis rate of positive and negative control groups was （36.10 ± 5.85 ）% and （7.87 ± 2.31 ）% respectively. After B16 melanoma ceils were treated with 7.68 mg/L UA for 24 h, the expression of MMP-9 protein was significantly decreased. The expression of MMP-9 protein in negative control group was 3. 106 folds more than the UA-PL-NP group, and 2.022 folds more than the positive control group. The expresstion of MMP-9 protein in the positive control group was 1. 537 folds more than the UA-PL-NP group. Conclusion UA-PL-NP can facilitate B16 melanoma cells apoptosis and inhibit proliferation of B16 melanoma cells cultured in vitro and restrain their metastasis to some extent.