目的：研究水蛭提取液对人视网膜母细胞瘤WERI-RB-1细胞的抑制作用。

方法：采用不同浓度水蛭提取液(0.02、0.04、0.08、0.16U/mL)作用于体外培养的WERI-RB-1细胞0、24、48、72h，经CCK-8法筛选最佳药物干预浓度和时间进行后续实验。将体外培养的WERI-RB-1细胞分为对照组(正常培养基培养)和实验组(含水蛭提取液培养基培养)，采用流式细胞仪检测药物对细胞周期和细胞凋亡的影响，Transwell侵袭实验检测药物对细胞侵袭能力的影响。

结果：根据CCK-8法检测结果选择0.04、0.08U/mL水蛭提取液作用48h为最佳干预条件进行实验。水蛭提取液干预的细胞主要阻滞在G2/M期，其中0.04、0.08U/mL实验组处于G2/M期的阳性细胞率分别为(12.59±5.36)%、(14.79±4.12)%，均明显高于对照组\〖(3.00±2.32)%，P<0.01\〗。水蛭提取液可诱导细胞凋亡，其中0.04、0.08U/mL实验组细胞凋亡率分别(37.91±3.44)%、(33.05±2.25)%，均明显高于对照组\〖(4.64±2.56)%，P<0.01\〗。Transwell侵袭实验检测结果显示，实验组Transwell小室下细胞数明显少于对照组，表明水蛭提取液能够抑制细胞侵袭。

结论：水蛭提取液在体外实验中能够抑制人视网膜母细胞瘤细胞的增殖、侵袭，并诱导细胞凋亡。

METHODS: WERI-RB-1 cells with different concentrations of leech extracts(0.02, 0.04, 0.08 and 0.16U/mL)were cultured in vitro for 0, 24, 48 and 72h. The optimal drug intervention concentration and time were selected by CCK-8 method for subsequent experiments. WERI-RB-1 cells cultured in vitro were divided into control group(normal culture medium)and experimental group(culture medium containing leech extract). Flow cytometry was used to detect the effects of drugs on cell cycle and cell apoptosis, and Transwell invasion assay was used to detect the effects of drugs on cell invasion abilit.

RESUITS: According to the detection results of CCK-8 method, the optimal intervention conditions were 0.04 and 0.08U/mL leech extract for 48h. The cells in the leech extract intervention were mainly blocked at G2/M stage, and the positive cell rates in the 0.04 and 0.08U/mL groups were(12.59±5.36)% and(14.79±4.12)%, respectively, which were significantly higher than those in the control group \〖(3.00±2.32)%, P<0.01\〗. The apoptosis rate of the cells in the 0.04 and 0.08U/mL groups was(37.91±3.44)% and(33.05±2.25)%, respectively, which were significantly higher than that in the control group \〖(4.64±2.56)%, P<0.01\〗. The results of Transwell invasion assay showed that the number of cells under the Transwell chamber in the experimental group was significantly than that in the control group, indicating that the leech extract could inhibit cell invasion.

CONCLUSION: The leech extract can inhibit the proliferation and invasion of human retinoblastoma cells and induce cell apoptosis in vitro.