紫龙金抑制人乳腺癌细胞系MCF-7生长的体内外实验研究

目的探讨复方中药紫龙金在体内和体外对MCF-7人乳腺癌细胞生长的抑制作用及其作用机制。方法 (1)台盼蓝拒染法和双层软琼脂克隆形成法检测紫龙金对MCF-7细胞增殖活性和克隆形成的影响;(2)Westernblot法检测紫龙金对p-ERK1/2蛋白表达的影响;(3)17-β-雌二醇缓释药片包埋法构建MCF-7细胞的BALB/c裸鼠皮下移植瘤模型,并检测紫龙金(实验组灌胃生药20g紫龙金/kg体重)对肿瘤生长的抑制效应。结果 (1)紫龙金降低MCF-7细胞的存活率,并呈剂量和时间依赖性;(2)紫龙金组MCF-7细胞克隆形成率显著低于对照组,且随着浓度的增加,克隆形成能力明显降低0.75、1.5、3.0、6.0mg/mL紫龙金组细胞克隆形成抑制率分别为(12.66±1.54)%、(88.83±2.13)%、100%、100%],克隆直径也明显减小或无克隆形成;(3)紫龙金下调p-ERK1/2蛋白的表达;(4)实验组裸鼠与对照组裸鼠4周后移植瘤的平均体积分别为(0.73±0.58)cm3、(1.36±0.64)cm3,平均瘤重分别为(1.02±0.25)g、(1.66±0.09)g,两组比较,差异均有统计学意义(P0.05),实验组抑瘤率为38.55%。结论紫龙金可显著抑制MCF-7细胞的体内和体外增殖、转化和致瘤性,这可能与p-ERK1/2蛋白的表达下调有关。

Experimental Study on Suppressive Effect of Zilongjin on Growth of Human Breast Cancer Cell Line MCF-7 in Vitro and in Vivo

Objective To investigate the suppressive effect and mechanism of action of Zilongjin （ZLJ,a composite Chinese drug） on the growth of human breast carcinoma cell line MCF-7 in vivo and in vivo.Methods The cell survival rate and clone forming efficiency were observed by direct cell counting with trypan blue staining and double layers soft agar test;the p-ERK 1/2 expression was analyzed using Western blotting;17-β-estrogen pellet embedding was adopted to make the subcutaneous transplanted tumor MCF-7 cell in BALB/c nude mice for detecting the tumor growth suppressive effect of ZLJ （20g crude drug/kg）.Results The survival rate of MCF-7 cell was obviously decreased by ZLJ in a time and dose-dependent manner;only few and small clones on soft agar could be found after treated with ZLJ,the inhibition rates of clone formation（%） for 0.75,1.5,3,6 mg/mL of ZLJ were 12.66±1.54,88.83±2.13,100 and 100,respectively,as compared with that of non-treated.The expression of p-ERK 1/2 was suppressed and the ability of the tumorigenicity in nude mice was reduced effectively by ZLJ.The mean volumes and weights of tumor in the test group and the control group were （0.73±0.58）cm3 vs （1.36±0.64）cm3 and （1.02±0.25）g vs （1.66±0.09）g respectively,showing significant difference （P0.05）,and the tumor inhibition rate of ZLJ was 38.55%.Conclusion ZLJ shows obviously suppresive actions on malignant proliferation,transformation and tumorigenicity of human breast carcinoma cell line MCF-7;the down-regulation of p-ERK 1/2 protein may involve in these effects.