基于Wnt/β-catenin信号通路探讨复方土贝母对裸鼠乳腺癌移植瘤的影响

Effects of Compound Tubeimu on Breast Cancer Xenografts in Nude Mice Based on Wnt/β-catenin Signaling Pathway

[目的]研究复方土贝母对裸鼠乳腺癌移植瘤生长以及对Wnt/β-catenin信号通路相关蛋白的影响。[方法]40只4～5周龄雌性BALB/C裸鼠皮下接种1×10~7/mL MCF-7细胞悬液0.2mL/只,造模后随机分为荷瘤对照组、复方土贝母低、中、高剂量组。复方土贝母各剂量组分别以0.5、1.0、2.0g·m L^(-1)的复方土贝母连续灌胃30d,对照组予0.9%氯化钠溶液灌胃,灌胃容积均为0.6mL/20g。所有小鼠第30天处死,测量各组移植瘤体积,并计算抑瘤率。实时荧光定量PCR检测β-连环蛋白(β-catenin)、E-钙粘蛋白(E-cadherin)、波形蛋白(Vimentin)mRNA表达水平,Western blot和免疫组化检测β-catenin、E-cadherin、Vimentin蛋白表达水平。[结果]复方土贝母低、中、高剂量组抑瘤率分别为10.56%、37.06%、33.17%,中、高剂量组移植瘤体积小于荷瘤对照组,差异有统计学意义(P<0.05),复方土贝母各剂量组的β-catenin、Vimentin m RNA表达低于荷瘤对照组,E-cadherin mRNA表达高于荷瘤对照组,差异有统计学意义(P<0.05)。中、高剂量组β-catenin、Vimentin蛋白表达低于荷瘤对照组,E-cadherin蛋白表达高于荷瘤对照组,差异有统计学意义(P<0.05)。[结论]复方土贝母能抑制乳腺癌移植瘤的生长,可能与抑制肿瘤中β-catenin、Vimentin的表达,上调E-cadherin表达有关。

[Objective]To study the inhibitory effect of compound Tubeimu on MCF-7 xenografts and the effects of Wnt/β-catenin signaling pathway-related proteins.[Methods]Forty female BALB/C nude mice aged 4 to 5 weeks were subcutaneously inoculated with 1×107/mL MCF-7 cell suspension0.2 mL each.After modeling,they were randomly divided into tumor control group,compound Tubeimu low,medium and high dose group.The drug-administered group was continuously administered with 0.5 g·mL-1,1.0 g·mL-1,2.0 g·mL-1 compound Tubeimu for 30 days,and tumor control group was given normal saline,and the gastric volume was 0.6 mL/20 g.All mice were sacrificed on the 30 th day,the tumor volume of each group was measured,and the tumor inhibition rate was calculated.The expression ofβ-catenin,E-cadherin,Vimentin m RNA was detected by fluorescent quantitative PCR,and protein expression was detected by Western blot and immunohistochemistry.[Results]The tumor inhibition rates of compound Tubeimu low,medium and high dose groups were 10.56%,37.06%and 33.17%,respectively.The tumor volume of medium and high dose groups was significantly different from that of tumor control group(P<0.05).The mRNA expression ofβ-catenin,Vimentin in compound Tubeimu groups was lower than tumor control group(P<0.05),while mRNA expression of E-cadherin in compound Tubeimu groups was higher than tumor control group(P<0.05).The protein expression ofβ-catenin,Vimentin in compound Tubeimu medium and high groups was lower than tumor control group(P<0.05),while protein expression of E-cadherin in compound Tubeimu medium and high groups was higher than tumor control group(P<0.05).[Conclusion]Compound Tubeimu can inhibit the growth of breast cancer xenografts,which may be related to inhibit the expression ofβ-catenin and Vimentin,and upregulate the expression of E-cadherin.