苏拉明联合人参皂苷Rg3抑制肺腺癌小鼠移植瘤转移的作用及机制

Inhibitory effects of suramin in combination with PG-Rg3 on the metastasis of lung adencarcinoma in mice

目的血管生成抑制剂靶向治疗肿瘤已成为目前研究热点之一。本研究旨在观察苏拉明联合人参皂苷Rg3(PG-Rg3)对肺腺癌小鼠异体移植瘤转移的协同抑制作用,并初步探讨苏拉明和PG-Rg3抑制肿瘤转移的相关机制。方法复制高转移肺腺癌小鼠移植瘤模型,将40只接种高转移性Lewis肺癌的C57BL/6J小鼠按随机数字表法随机分成5组:对照组、顺铂组、苏拉明组、PG-Rg3组及苏拉明+PG-Rg3组(联合组)。给予相应药物干预,于接种后24 d处死各组小鼠,剥离皮下肿瘤和取出双肺,计算肺转移发生情况,免疫组化和图像分析系统检测皮下移植瘤中组织蛋白酶B(CB)和血管内皮生长因子(VEGF)表达并定量分析,RT-PCR半定量检测组织蛋白酶B mRNA、VEGF mRNA表达。结果对照组、顺铂组、苏拉明组、PG-Rg3组、联合组的肺表面转移结节数分别为(14.00±2.90)、(9.77±1.65)、(6.00±1.70)、(5.80±2.02)、(1.93±1.12),CB灰度值分别为(147.24±5.51)、(129.57±5.41)、(87.39±5.03)、(81.38±5.31)、(67.36±4.37),CB mRNA相对量分别为(85.07±9.24)、(80.45±9.01)、(45.21±7.53)、(39.67±6.84)、(24.33±6.10),VEGF灰度值分别为(156.26±6.45)、(134.7±5.81)、(96.13±5.66)、(90.80±4.92)、(68.73±4.31),VEGF mRNA相对量分别为(91.07±9.43)、(84.45±9.50)、(49.67±8.07)、(45.48±6.46)、(27.39±5.19),苏拉明组和PG-Rg3组中肺表面转移结节数、皮下肿瘤内CB、CB mRNA、VEGF、VEGF mRNA表达与对照组相比明显下降,相反顺铂组对此则无明显影响。联合用药组则显著抑制肿瘤肺表面转移结节数且具有协同作用。结论苏拉明和PG-Rg3对肺腺癌移植瘤的转移具有协同抑制作用,其机制与其抑制肿瘤内CB、CB mRNA、VEGF、VEGF mRNA表达相关。

[ Objective ] The scheme of tumor therapy through angiogenesis becomes one of studying hot spots at present. The experiment is designed to observe that if suramin combination with SPG-Rg3 has synergetic effects of suppressing lung adenocareinoma＇s metastasis and to study the related mechanism of suramin and PG-Rg3＇s sup- pressing tumor metastasis. [ Methods ] Forty C57BL/6J mice bearing high metastatic Lewis cells were randomized in- to five groups： control group; DDP group; suramin group; PG-Rg3 group; combination group. Each group to be the appropriate intervention, all the mice was sacrificed by tumor inoculation in 24 days, Lung and subcutaneous tumors were dressed for histological examination. The metastatic tumor foci on lung surface in mice were observed. Lung metastatic occurring rate and metastatic foci inhibitory rate were counted. CB and VEGF were determined by im- munohistochemistry method with image analyze system, and detection of PGPmRNA, MRPlmRNA expression byRT-PCR. [Results] Control group, cisplatin group, suramin group, SPG-Rg3 group, combined group of lung metastatic nodules were （14.00±2.90）, （9.77±1.65）, （6.00±1.70）, （5.80±2.02）, （1.93±1.12）, the grey of CB were （147.24±5.51）, （129.57±5.41）, （87.39±5.03）, （81.38±5.31）, （67.36±4.37）, the relative amount of CBmRNA respec- tively were （85.07±9.24）, （80.45±9.01）, （45.21±7.53）, （39.67±6.84）, （24.33±6.10）, the grey of VEGF were （156.26± 6.45）, （134.7±5.81）, （96.13±5.66）, （90.80±4.92）, （68.73±4.31）, the relative amount of VEGFmRNA were （91.07± 9.43）, （84.45±9.50）, （49.67±8.07）, （45.48±6.46）, （27.39±5.19）. In the Suramin group and PG-Rg3 group, the number of lung metastatic nodules, the expression of CB, CBmRNA, VEGF, VEGFmRNA in subcutaneous tumor was signifi- cantly decreased compared with the control group, contrary to this, cisplatin group had no significant effect. Combi- nation group was significantly inhibited on lung metastatic nodules of tumor, it has a synergistic effect. [ Conclu- sions ] Suramin combination with PG-Rg3 can produce a synergetic inhibitory activity against lung metastasis of lung adenocarcinoma. The fact that suramin and PG-Rg3 inhibited lung metastasis of lung adenoearcinoma may be associ- ated its effect of suppressing the expression of CB, CBmRNA, VEGF, VEGFmRNA in tumor tissue.