西黄丸调节MEKK1/SEK1通路抑制小鼠乳腺癌生长机制研究

Xihuang Pill Inhibit Growth of Mouse Breast Cancer by Regulating MEKK1/SEK1 Pathway

目的研究西黄丸抑制4T1乳腺癌荷瘤小鼠肿瘤的生长及其可能的发生机制。方法建立4T1乳腺癌荷瘤小鼠模型,以西黄丸低、中、高剂量ig给药14 d,剥离肿瘤组织称重,TUNEL染色检测肿瘤细胞的凋亡,Real-time PCR法检测肿瘤组织中MEKK1和SEK1 mRNA表达,免疫荧光染色法和Western Blot法检测肿瘤组织中MEKK1和SEK1蛋白表达。结果与阴性对照组比较,给药组肿瘤体积及质量随西黄丸剂量的升高而降低;TUNEL染色显示凋亡的肿瘤细胞增加;Real-time PCR显示肿瘤组织中MEKK1和SEK1 mRNA表达升高;免疫荧光染色和Western Blot显示肿瘤组织中MEKK1和SEK1蛋白表达水平上调;以上结果均与西黄丸呈剂量依赖性,P <0.05具有统计学差异。结论西黄丸可能通过促进4T1乳腺癌荷瘤小鼠肿瘤细胞凋亡抑制肿瘤的生长,其机制可能与西黄丸上调4T1乳腺癌荷瘤小鼠肿瘤组织中MEKK1、SEK1 mRNA和蛋白表达有关。

Objective To study the effect of Xihuang Pill on the growth of 4T1 mouse breast cancer and its possible mechanism. Methods A mouse 4T1 breast cancer model has been established. Model mice are administered Xihuang Pill at low, medium and high dose for 2 weeks meanwhile tumor growth curve is drawn, and tumor tissues are then removed and weighed. Tumor cell apoptosis is detected by Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling staining(TUNEL), mRNA expression levels of Mitogen-activated protein kinase kinase kinase 1(MEKK1) and SAPK/Erk kinase 1(SEK1) in tumor tissue are detected by Real-time PCR and their protein expression levels is detected by immunofluorescence staining and Western Blot. ResultsComparing with the negative control group, tumor volumes and tumor weights in the Xihuang Pill groups decreas significantly with increasing doses of Xihuang Pill. TUNEL staining show that the number of apoptotic tumor cells increas with increasing doses of Xihuang Pill. Real-time PCR show that mRNA expression of MEKK1 and SEK1 in tumor tissue increase with increasing doses of Xihuang pill. Immunofluorescence staining and Western Blotting show that protein expression of MEKK1 and SEK1 in tumor tissue increase with increasing doses of Xihuang pill. Conclusion Xihuang Pill might promote tumor cell apoptosis and further inhibit the tumor growth of 4T1 mouse breast cancer. The mechanism of Xihuang Pill may be related to upregulation of mRNA and protein expression of MEKK1 and SEK1 in tumor tissue of 4T1 mouse breast cancer.