穿膜融合多肽TAT-N24对S180腹水瘤生长的抑制作用

Inhibitory Effects of the Cell-permeable TAT-N24 Fusion Peptide on the Growth of S180 Ascites Tumor

目的观察穿膜融合多肽TAT-N24对S180腹水瘤生长的抑制作用。方法建立S180腹水瘤小鼠模型,以不同剂量TAT-N24腹腔内注射,观察TAT-N24对腹水瘤小鼠腹水生成的影响,流式细胞仪检测腹水瘤细胞细胞周期进程,应用5-溴脱氧尿嘧啶核苷(BrdU)掺入法检测TAT-N24对细胞DNA合成的影响,验证TAT-N24的抗肿瘤作用。结果腹水测量结果显示,模型对照组腹水为(8.3±2.3)mL,实验组分别为:TAT-N245μL组(3.2±1.2)mL,TAT-N2420μL组(2.7±1.0)mL,TAT-N24100μL组(1.3±0.2)mL;结果提示给予腹腔注射TAT-N24能显著抑制腹水的生成(P<0.05);BrdU/PI双掺入法检测细胞DNA合成结果显示,对照组BrdU阳性细胞数占总细胞比例为(25.86±3.54)%,而给予TAT-N24高剂量(100μL)组BrdU阳性细胞数占总细胞比例为(5.91±0.57)%,2组间差异有统计学意义(P<0.01);对照组动物腹水瘤细胞中G0/G1期细胞为(63.88±4.01)%,S期和G2/M期细胞分别为(23.93±2.91)%和(12.19±1.62)%,而TAT-N24高剂量组动物腹水瘤细胞G0/G1期细胞增加至(83.71±1.53)%,S期和G2/M期细胞分别为(7.56±1.40)%和(8.72±0.73)%,2组比较差异有统计学意义(P<0.01)。结论融合多肽TAT-N24能有效抑制S180腹水瘤小鼠的腹水生成,阻滞腹水瘤细胞细胞周期进程,抑制细胞DNA合成。

Objective To elucidate the inhibitory effects of the cell-permeable TAT-N24 fusion peptide on the growth ability of S180 ascites tumor.Methods The murine S180 ascites tumor model was established.Different doses of TAT-N24 were injected intraperitoneally,and the effects of TAT-N24 on the ascites formation were observed.The cell cycle of S180 ascites tumor was assayed by flow cytometry.5-bromodeoxyuridine（BrdU/PI）incorporation method was applied to detect DNA synthesis in the tumor cells to clarify the anti-tumor role of TAT-N24.Results In model control group,the volume of ascites was（8.3±2.3）mL,and that in the experiment groups was（3.2±1.2）mL（TAT-N24 5 μL）,（2.7±1.0）mL（TAT-N24 20 μL）,（1.3±0.2）mL（TAT-N24 100 μL）,respectively.In the control group the percentage of BrdU positive cells was（25.86±3.54）%,and that in the experiment group（TAT-N24 100 μL）was（5.91±0.57）% with the significant difference between two groups（P〈0.01）.Cell cycle analysis revealed that the proportion of cells in G0/G1,S and G2/M phases was（63.88±4.01）%,（23.93±2.91）% and（12.19±1.62）% respectively in control group.However in TAT-N24 group,the proportion of cells in G0/G1,S and G2/M phases was（83.71±1.53）%,（7.56±1.40）% and（8.72±0.73）% respectively.There was significant difference between TAT-N24 group and control group（P〈0.01）.Conclusion TAT-N24 fusion peptide can induce cell cycle arrest,inhibit the DNA synthesis of tumor cells and suppress tumor growth in murine S180 ascites tumor model.