药学学报, 2019, 54(11): 2011-2018
引用本文:
张铭予, 顾琼, 周晓伟, 李莹, 张远远, 张玉君, 王伟华, 李春雨, 李国辉. 基于转移前微环境的四君子汤提高吉非替尼疗效研究[J]. 药学学报, 2019, 54(11): 2011-2018.
ZHANG Ming-yu, GU Qiong, ZHOU Xiao-wei, LI Ying, ZHANG Yuan-yuan, ZHANG Yu-jun, WANG Wei-hua, LI Chun-yu, LI Guo-hui. Sijunzi Tang improves the efficacy of gefitinib through pre-metastatic niche[J]. Acta Pharmaceutica Sinica, 2019, 54(11): 2011-2018.

基于转移前微环境的四君子汤提高吉非替尼疗效研究
张铭予, 顾琼, 周晓伟, 李莹, 张远远, 张玉君, 王伟华, 李春雨, 李国辉
国家癌症中心, 国家肿瘤临床医学研究中心, 中国医学科学院、北京协和医学院肿瘤医院, 北京 100021
摘要:
利用Lewis肺癌(LLC)转移小鼠模型,研究吉非替尼(gefitinib)与四君子汤(Sijunzi Tang,SJZ)对转移前微环境影响。实验方案经中国医学科学院肿瘤医院伦理委员会审议同意并批准。首先将1×106个荧光素酶标记的LLC细胞接种于小鼠右后腋下,构建肺癌转移模型。将小鼠按体重随机分为模型组、吉非替尼组(50 mg·kg-1)、SJZ组(25.74 g·kg-1)和联合给药组,造模后次日开始给药。连续给药14天后,利用小动物成像技术检测肿瘤大小;通过流式细胞技术检测小鼠外周血、脾、肺组织中单核细胞和中性粒细胞数量及其表面趋化因子受体(CXCR1、CCR2)和致癌基因c-Kit的表达水平变化情况;使用酶联免疫吸附测定(ELISA)法检测小鼠血浆及瘤组织中炎症因子(IL-1α、IL-6)的含量;给药21天后,取肿瘤组织,称重,采用游标卡尺测量肿瘤体积大小并评估联合给药的抑瘤效果;给药45天后,记录各组小鼠的生存期。流式细胞检测结果显示,在小鼠肺组织中,与模型组比较,吉非替尼组、SJZ组和联合给药组的中性粒细胞的百分比均呈显著性下降(P<0.05或P<0.01),但三组之间比较无显著性差异(P>0.05);在小鼠外周血和肺组织中,与模型组比较,SJZ组与联合给药组的中性粒细胞和单核细胞表面CXCR1、CCR2和c-Kit表达水平均呈下降趋势(P<0.05或P<0.01),但吉非替尼组单核细胞表面c-Kit表达水平呈显著性上升(P<0.05);在小鼠脾组织中,吉非替尼组CXCR1、CCR2和c-Kit表达水平均呈上升趋势(P<0.05),而SJZ组和联合给药组均呈下降趋势(P<0.05或P<0.01)。ELISA法检测结果显示,在小鼠血浆中,与模型组比较,SJZ组IL-1α含量呈显著性下降(P<0.01),联合给药组IL-6含量呈显著性下降(P<0.05);与吉非替尼组比较,联合给药组IL-1α含量显著性下降(P<0.05)。在小鼠肿瘤组织中,与模型组比较,联合给药组IL-1α含量呈显著性下降(P<0.05)。与吉非替尼组比较,联合给药组IL-1α、IL-6含量与SJZ组IL-6含量均呈显著性下降(P<0.05)。连续给药21天后,吉非替尼组、SJZ组和联合给药组的抑瘤率分别为45.7%、38.4%和84.8%。给药45天后,模型组生存率为0%,吉非替尼组、SJZ组和联合给药组的生存率分别为40%、60%和60%。上述结果表明,SJZ可通过调控转移前微环境提高吉非替尼的疗效。
关键词:    肺癌      吉非替尼      四君子汤      肿瘤转移      转移前微环境      炎性细胞因子     
Sijunzi Tang improves the efficacy of gefitinib through pre-metastatic niche
ZHANG Ming-yu, GU Qiong, ZHOU Xiao-wei, LI Ying, ZHANG Yuan-yuan, ZHANG Yu-jun, WANG Wei-hua, LI Chun-yu, LI Guo-hui
National Cancer Center, National Clinical Research Center for Cancer, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China
Abstract:
The Lewis lung carcinoma (LLC) metastatic mouse model was used to investigate the effects of gefitinib and Sijunzi Tang (SJZ) on pre-metastatic niche. The experimental protocol was approved by the Ethics Committee which belongs to Cancer Hospital, Chinese Academy of Medical Sciences. To generate spontaneous lung metastatic models, 1×106 luciferase-labeled LLC cells were injected subcutaneously in the shaved right flank of mice. One day after LLC inoculation, the mice were randomly divided into model (saline), gefitinib (50 mg·kg-1) treatment, SJZ treatment (25.74 g·kg-1), and co-treatment gefitinib with SJZ groups, with intragastrical administration. After 14 days of continuous administration, tumor size was detected by IVIS® Spectrum system. The number of monocytes and neutrophils and the expression levels of chemokine receptors (CXCR1, CCR2) and carcinogenic gene (c-Kit), in peripheral blood, spleen and lung tissues of mice were determined by flow cytometry. The contents of interleukin-IL-1α (IL-1α) and interleukin-6 (IL-6) were detected by the enzyme linked immunosorbent assay (ELISA). After 21 days of treatment, tumors were surgically removed, weighed and the tumor volume was measured with vernier caliper and the antitumor effect of co-administration was evaluated. After 45 days of administration, the survival of mice was recorded. The results of flow cytometry showed that the percentage of neutrophils in gefitinib group, SJZ group, and co-treatment group was significantly decreased in the lung tissue compared to the model group (P<0.05 or P<0.01), but there was no significant difference between three treatment groups (P>0.05). In the mouse peripheral blood and lung tissue, compared with the model group, the expression levels of CXCR1, CCR2 and c-Kit on the surface of neutrophils and monocytes in SJZ group and co-treatment group decreased or decreased significantly (P<0.01 or P<0.05). However, there was a significant increase in the expression level of c-Kit on the surface of monocytes (P<0.05). In the mouse spleen tissue, the expression levels of CXCR1, CCR2 and c-Kit in the gefitinib group increased significantly (P<0.05), while decreased significantly in SJZ or co-treatment group (P<0.05). The results of ELISA showed that the content of IL-1α in SJZ group decreased significantly in the plasma of the mice compared with the model group (P<0.01) and the content of IL-6 in co-treatment group decreased significantly (P<0.05). Compared with the gefitinib group, the content of IL-1 in the co-treatment group decreased significantly (P<0.05). In the tumor tissues of mice, compared with the model group, the content of IL-1α in the co-treatment group decreased significantly (P<0.05). Furthermore, the content of IL-1α in co-administrated group and IL-6 in SJZ or co-treatment group decreased significantly compared with the gefitinib group (P<0.05). After 21 days of continuous administration, the tumor inhibition rates of gefitinib group, SJZ group and co-administrated group were 45.7%, 38.4%, and 84.8%, respectively. After 45 days of administration, the survival rate of the model group was 0%, whereas the gefitinib, SJZ or co-treatment group has a survival rate of 40%, 60%, or 60%, respectively. In summary, our study illustrated that Sijunzi Tang could improve the anti-tumor effect of gefitinib by regulating pre-metastatic niche.
Key words:    lung cancer    gefitinib    Sijunzi Tang    tumor metastasis    pre-metastatic niche    inflammatory cytokine   
收稿日期: 2019-06-21
DOI: 10.16438/j.0513-4870.2019-0502
基金项目: 中国医学科学院医学与健康科技创新工程重大协同创新项目(2016-I2M-1-001).
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