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[摘要]
目的 研究西黄丸抑制荷4T1小鼠乳腺癌肿瘤的生长及其可能机制。方法 建立荷4T1小鼠乳腺癌模型,以低、中、高剂量(0.39、0.78、1.95 g/kg) ig给予西黄丸两周,分离肿瘤组织,称质量并计算抑瘤率;切片,TUNEL染色检测肿瘤细胞凋亡;实时荧光定量PCR (qRT-PCR)检测肿瘤组织JNK1、AP-1 mRNA表达;免疫荧光染色和Western Blotting检测肿瘤组织中JNK1、AP-1蛋白表达。结果 与模型组比较,肿瘤质量随西黄丸剂量的升高显著下降;TUNEL荧光染色结果显示,肿瘤细胞凋亡数量随西黄丸剂量的升高显著升高;qRT-PCR结果显示,肿瘤组织中JNK1、AP-1 mRNA表达水平随西黄丸剂量的升高显著上调;免疫荧光染色和Western Blotting结果显示,肿瘤组织中JNK1、AP-1蛋白表达随西黄丸剂量的升高显著增加;且高、中、低剂量组差异均具有统计学意义(P<0.05)。结论 西黄丸显著抑制肿瘤生长,其作用机制与激活JNK1/AP-1信号通路促进肿瘤细胞凋亡相关。
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[Abstract]
Objective To study the mechanism of Xihuang Pills on growth inhibition of 4T1 breast cancer cells in mice.Methods The model of 4T1 breast cancer mice was established. After administrating Xihuang Pills at low-, medium-, and high-dose level (0.39, 0.78, and 1.95 g/kg) for two weeks, tumor tissue was weighed, sliced, and homogenized. Tumor cell apoptosis was detected by TUNEL staining. The expression of mRNAs in JNK1 and AP-1 in tumor tissue was detected by Real-time quantitative PCR. The expression of JNK1 and AP-1 protein in tumor tissue was detected by immunofluorescence staining and Western Blotting.Results Comparing with model group, the tumor weights of Xihuang Pills group decreased significantly with the increase of doses. TUNEL staining results showed that the number of apoptosis of tumor cells increased with the increase of doses of Xihuang Pills. Real-time PCR results showed that the expression of JNK1 and AP-1 mRNA in tumor tissue increased with the increase of doses of Xihuang Pills. The results of immunofluorescence staining and Western Blotting showed that the expression of JNK1 and AP-1 protein in tumor tissue increased with the increase of doses of Xihuang Pills. The differences in the high, medium and low dose groups were statistically significant (P< 0.05).Conclusion Xihuang Pills significantly inhibit the growth of tumor cells, which mechanism is related to the activation of JNK1/AP-1 signaling pathway, with view to promoting the apoptosis of tumor cells.
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[基金项目]
国家自然科学基金资助项目(81573664)