摘要
目的:研究益肾解毒方对小鼠溶骨性骨转移瘤细胞核糖核酸酶L(ribonuclease L,RNase L)蛋白及凋亡终末蛋白半胱氨酸蛋白酶-3(Caspase-3)蛋白表达的调控作用。方法:将已经获得国家专利的肺腺癌溶骨性骨转移细胞株,给予裸鼠左心室注射,制备溶骨性骨转移模型,随机分为5组,益肾解毒方高、中、低剂量(24,12,3 g·kg^(-1))组,帕米磷酸二钠组及模型组,以Micro SPECT/CT显像及病理组织学方法观察骨转移发生情况,定时定量给药并于5周后取材,通过免疫组织化学、蛋白免疫印迹法(Western blot)以及反转录聚合酶链式反应(RT-PCR)等方法研究RNase L,Caspase-3及益肾解毒方的干预作用。结果:与模型组比较,帕米磷酸二钠组、益肾解毒方高、中、低剂量组骨转移瘤细胞RNase L蛋白,mRNA表达降低(P<0.01),益肾解毒方中、低剂量组及帕米磷酸二钠组骨转移灶中Caspase-3蛋白,mRNA表达升高(P<0.01)。讨论:肺腺癌骨转移灶中RNase L高表达,而细胞凋亡过程中最主要的终末剪切酶Caspase-3表达降低,其内在机制可能与肺腺癌骨转移细胞的凋亡逃避机制相关,益肾解毒方能够降低RNase L表达,增强Caspase-3表达,一定程度上抑制了肿瘤细胞的凋亡逃避,这可能是益肾解毒方抗肿瘤的内在机制。
Objective： To explore the regulating effect of Yishen Jiedu prescription on the protein expression of ribonuclease L（ RNase L） and the apoptosis terminal protein Caspase-3 in bone metastasis from lung adenocarcinoma. Method： Lung adenocarcinoma cell lines that had won the national patent were injected into left ventricle of nude mice to prepare the bone metastasis model. Then,the mice were randomly divided into five groups： high,middle and low-dose（ 24,12,3 g·kg^（-1）） Yishen Jiedu groups,disodium phosphate group and control group. Finally,with histopathological method and micro SPECT/CT imaging method,we can observe the outcome of bone metastases. After administration at fixed time and dose for 5 weeks,dissection was performed,and the expressions of RNase L protein and Caspase-3 protein in bone metastasis from lung adenocarcinoma,and the regulatory effect of Yishen Jiedu prescription was analyzed through immunohistochemistry methods,Western blot and reverse transcription polymerase chain reaction（ RT-PCR） methods. Result： The model group had the highest expression of RNase L protein and mRNA in bone metastases tumor cells,with significant statistical differences between the model group and others（ P〈0. 01）. However,the model group showed the lowest expression of Caspase-3 protein and mRNA,with significant statistical differences between the middle low dosage group of Yishen Jiedu and disodium phosphate groups（ P〈0. 01）. Conclusion： There was a high expression in RNase L expression in bone metastases in patients with lung adenocarcinoma,while Caspase-3,a main terminal protein in the process of cell apoptosis,was extremely low. The internal mechanism may be correlated with the cell apoptosis escape of lung adenocarcinoma bone metastases. The prescription of Yishen Jiedu prescription can inhibit the apoptosis of tumor escape by reducing the expression of RNase L and enhancing caspase-3 protein.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第20期114-119,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中国中医科学院中央级公益性科研院所科研基本业务费自主选题项目(ZZ070805)
