3TSR对胆囊癌的抑制作用及初步机制

Inhibitory effects of 3TSR on gallbladder cancer and its rudimentary mechanism

目的研究血管形成抑制剂3TSR对裸鼠皮下胆囊癌种植瘤的抑制作用及其机制。方法建立裸鼠皮下胆囊癌种植瘤模型,进行随机干预实验,实验分为对照组、3TSR组、5-氟尿嘧啶(5-FU)组和3TSR+5-FU组,每组6只。检测荷瘤体积、胆囊癌细胞增殖指数(PCNA)、凋亡率、微血管密度(MVD)、肿瘤血管内皮细胞凋亡率及血管内皮生成因子(VEGF)的表达。结果(1)3TSR组、5-FU组和3TSR+5-FU组肿瘤体积较对照组显著减小,分别缩小31.1%,38.3%和60.2%。对照组、3TSR组、5-FU组和3TSR+5-FU组胆囊癌细胞增殖指数分别为(25.8±2.4)%,(23.7±5.0)%,(9.4±2.5)%和(11.9±3.8)%,其中3TSR组增殖指数与对照组比较无显著差异,但5-FU组和3TSR+5-FU组细胞增殖指数显著小于对照组和3TSR组(P<0.05)。4组胆囊癌细胞凋亡率分别为(5.1±1.4)%,(4.3±1.6)%,(19.8±5.1)%和(13.9±3.8)%,其中3TSR组胆囊癌细胞凋亡率与对照组比较无显著差异(P<0.05),但5-FU组和3TSR+5-FU组细胞凋亡率显著大于对照组和3TSR组(P<0.05)。4组微血管密度分别为(15.2±4,6)%,(4.2±1.2)%,(8.7±2.1)%和(4.9±1.8)%,其中含3TSR的两组显著低于对照组和5-FU组(P<0.05)。4组内皮细胞凋亡率分别为(4.3±1.4)%,(10.8±2.9)%,(5.1±2.1)%和(10.2±3.0)%,其中含3TSR组显著高于对照组(P<0.05)。4组VEGF表达率分别为(14.0±3.2)%,(4.7±2.8)%,(12.8±1.4)%和(6.7±2.9)%,其中含3TSR的两组显著低于对照组和5-FU组(P<0.05)。结论3TSR显著减少肿瘤体积,抑制微血管生成.其机制可能与诱导内皮细胞凋亡和抑制VEGF有关;3TSR与化疗药物5-FU合用对胆囊癌的治疗有协同作用。本研究为3TSR的临床前期应用提供了一定实验基础。

Objective To investigate the anti-tumor efficacy of angiogenic inhibitor 3TSR on gallbladdercancer cells and explore its rudimentary mechanism. Methods Human gallbladder carcinoma cell line GBC-SD was inoculated subcutaneously into BALB/c mice and then the mice were divided into four groups： control group, 3TSR group, 5-FU group and 3TSR plus 5-FU group with 6 mice in each. The tumor volume, cancer cell proliferation, apoptosis rate, endothelial cell apoptosis rate and expression of vascular endothelial growth factor （VEGF） were detected, respectively. Results The tumor volume of treated groups was reduced by 31.1%,38.3% and 60.2%, respectively. The proliferation index（PCNA） of control group, 3TSR, 5-FU and 3TSR plus 5-FU groups was （25.8 ± 2.4） %, （23.7±5.0） %, （9.4±2.5） % and （11.9±3.8）, respectively. The proliferation index was not statistically different between 3TSR and control group, but that in 5-FU and 3TSR plus 5-FU group was significantly lower than that in control group （P〈0 05）. The cancer cell apoptosis rate of four groups was （5.1±1.4）%,（4.3±1.6）%, （19.8±5.1）% and （13.9±3.8）%, respectively. The apoptosis rate was not statistically different between 3TSR and control group, but that in 5-FU and 3TSR plus 5-FU group was significantly higher than that in 3TSR and control group （P〈005）. The average microvessel density was （ 1 5.2 ±4.6 ） %, （ 4.2 ± 1.2 ） %, （ 8.7 ± 2.1 ） % and （ 4.9 ± 1.8） %, respectively. The average microvessel density in 3TSR and 3TSR plus 5-FU group was significantly lower than that in control and 5-FU group （P〈005）. The endothelial cell apoptosis rate of four groups was （4.3±1.4） %, （10.8±2.9）%, （5.1 ±2. 1）% and （10.2±3.0）% ,respectively. The endothelial cell apoptosis rate in 3TSR and 3TSR plus 5-FU group was significantly higher than that in control and 5-FU group （P〈0. 05）. The positive rates of VEGF expression in the above four groups were （ 14.0±3.2） %, （4.7 ±2.8） %, （ 12.8 ± 1.4） % and （ 6.7 ±2.9） %, respectively, and it was markedly higher in 3TSR and 3TSR plus 5-FU groups than in the control and 5 FU group（P〈 0. 05）. Conclusions 3TSR has no direct inhibitory effects on the proliferation and apoptosis of gallbladder cancer, but it decrease the tumor volume significantly. The mechanism of anti-tumor effects of 3TSR may be mainly due to the induction of the apoptosis of endothelial cells and inhibition of the VEGF. 3TSR combined with 5-FU has additive effect on inhibitory activity against the growth of gallbladder cancer. This study provides some basic data for future pre-clinical and clinical development of 3TSR.