川芎嗪联合顺铂对小鼠Lewis肺癌中Mac2-BP和VEGF表达的影响

Effect of TMP combined with cisplatin on expression of Mac2-BP and VEGF in Lewis lung cancer mice

目的探讨川芎嗪(TMP)联合顺铂(DDP)对小鼠Lewis肺癌Mac2-结合蛋白(Mac2-BP)和血管内皮生长因子(VEGF)表达的影响。方法 C57BL/6小鼠皮下接种Lewis肺腺癌细胞,建立小鼠Lewis肺癌移植瘤模型,40只小鼠随机分为4组:生理盐水组(NS组,0.9%氯化钠,0.2 ml)、TMP组(TMP 100 mg/kg,0.2 ml)、DDP组(DDP2 mg/kg,0.2 ml)、TMP+DDP组(TMP 100 mg/kg,DDP 2 mg/kg;共0.2 ml),每组10只,于接种后14 d给予药物干预,每隔1 d测量瘤体长短径并计算肿瘤体积,连续用药14 d后处死小鼠,剥离皮下肿瘤称重并计算抑瘤率。采用Western印迹、免疫组化方法检测Mac2-BP、VEGF的表达。结果与NS组相比,TMP组、DDP组、TMP+DDP组的瘤体增长均减慢,TMP+DDP组瘤体增长减慢最明显。TMP组、DDP组、TMP+DDP组抑瘤率分别为25.57%、45.24%和66.5%。采用金氏公式评价TMP+DDP对小鼠Lewis肺癌的抑制作用,TMP+DDP组0.85<q<1.15,表明TMP+DDP对小鼠Lewis肺癌有相加抑制作用。NS组中,VEGF与Mac2-BP表达成正相关(Western印迹法r=0.713,P=0.021,免疫组化法r=0.653,P=0.041)。与NS组相比,Mac2-BP、VEGF在TMP组、DDP组、TMP+DDP组表达明显降低(P<0.05),TMP+DDP组表达最低(P<0.05)。两种检测方法结果一致。结论 TMP+DDP对小鼠Lewis肺癌有相加抑制作用,能抑制肿瘤血管生成,其机制可能与抑制Mac2-BP、VEGF表达有关。

[ Abstract] Objective To investigate the effect of tetramethylpyrazine （TMP） combined with cisplatin （DDP） on the expression of Mac2-binding protein（Mac2-BP） and vascular endothelial growth factor（VEGF） in mice with lung cancer. Methods C57BL/6 mice were subcutaneously inoculated with Lewis lung adenocarcinoma cells, and Lewis lung adenocareinoma mouse xenograft model was established. Forty mice were randomly divided into four groups： normal saline group（NS group ,0. 9% NaC1,0. 2 ml）, TMP group （TMP 100 mg/kg,O. 2 ml）, DDP group （DDP 2 mg/kg,0.2 ml）, TMP plus DDP group （doses as above ,0.2 ml totally） with 10 mice in each group. After 2 drug intervention, following 14 days of inoculation, the tumor diameter was measured every two days to calculate the tumor volume. The mice were sacrificed after 14 days of continuous medication, and the subcutaneous tumors were weighed after stripping to calculate the inhibitory rate. The expressions of Mac2-BP and VEGF was detected by Western blot and immunhistochemistry. Results Compared with NS group, the tumor growth rate of TMP group, DDP group and TMP ＋ DDP group was slowed down, and the tumor growth rate in the TMP ＋ DDP group was decreased most significantly. The inhibitory rate of TMP group, DDP group and TMP ＋ DDP group was 25.57% , 45.24% and 66.5% ,respectively. Kim＇s formula was used to evaluate the synergy of the combined treatment. The q values for TMP ＋ DDP group was 0.85 〈 q 〈 1. 15, suggesting the additive inhibition of combined TMP and DDP in mice Lewis lung cancer. A positive correlation was observed between Mac2-BP and VEGF expressions of NS group （ Western blotting r = 0. 713, P = 0. 021, immunohistochemistry r = 0. 653, P = 0. 041 ）. Compared with NS group, the expression of Mac2-BP and VEGF in the TMP group, DDP group, TMP ＋ DDP group was dramatically decreased （ P 〈 0.05 ） , and that in TMP ＋ DDP group was significantly lower than in TMP and DDP groups （P 〈 O. 05 ）. The results deduced by immunhistochemistry and Western blot were consistent. Conclusion Combination of TMP and cisplatin has additive anti-tumor effect on lung cancer bearing C57BL/6 mice. It can inhibit tumor angiogenesis and the mechanism may be related to the inhibition of Mac2-BP and VEGF expression. TMP can inhibit the growth of implanted Lewis lung carcinoma xenografted by inhibiting the expression of Mac2-BP and VEGF and the angiogenesis. TMP combined with DDP can improve the effect of chemotherapy and enhance the inhibiting effect of angiogenesis. Mac2-BP may promote the angiogenesis of lung cancer by up-regulating the expression of VEGF.