扶肺煎抑制肺癌增殖和转移的体内外研究  

In vivo and In vitro Study on Fufei Jian Inhibiting Proliferation and Metastasis of Lung Carcinoma

作　　者：陈观平[1] 汪一帆[1] 应栩华[1] CHEN Guan-Ping;WANG Yi-Fan;YING Xu-Hua(Cancer Institute of Integrated Chinese and Western Medicine,Zhejiang Provincial Litongde Hospital,Hangzhou 310012 Zhejiang,China)

机构地区：[1]浙江省立同德医院中西医结合肿瘤研究所,浙江杭州310012

出　　处：《广州中医药大学学报》2021年第5期1009-1017,共9页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：浙江省基础公益研究计划项目(编号:LGF18H290003);浙江省中医药科技计划资助项目(编号:2017ZB017);浙江省中医药科技计划项目(编号:2019ZB028)。

摘　　要：【目的】观察扶肺煎对肺癌增殖和转移的抑制作用。【方法】(1)体外研究:制备扶肺煎含药血清,以信号转导与转录激活因子(STAT3)抑制剂AG490、顺铂为阳性对照药,培养人肺癌A549、SPC-A1细胞,采用细胞计数试剂盒8(CCK8)法测定细胞增殖情况,细胞划痕实验观察细胞迁移能力,Transwell实验观察细胞体外侵袭能力,蛋白免疫印迹法检测细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白依赖性激酶4(CDK4)、STAT3、磷酸化STAT3(p-STAT3)蛋白的表达。(2)体内研究:取48只SPF级雄性C57BL/6小鼠,建立Lewis肺癌移植瘤模型,给予不同剂量扶肺煎、顺铂(阳性对照药)灌胃,第15天处死小鼠并剥离肺脏观察转移灶情况,将移植瘤称质量,计算抑瘤率及肺转移抑制率,并采用免疫组织化学法检测移植瘤组织Cyclin D1、CDK4、p-STAT3表达。【结果】扶肺煎组的A549、SPC-A1细胞增殖能力、迁移及侵袭能力均低于空白对照组,接近AG490及顺铂处理水平。在体内,扶肺煎高剂量组、顺铂组的抑瘤率分别为26.67%、52.53%,肺转移抑制率分别达到了28.76%、33.82%,且各给药组p-STAT3、Cyclin D1、CDK4表达水平均有所下降(P <0.05或P <0.01)。【结论】扶肺煎可通过抑制STAT3蛋白的活化,降低Cyclin D1、CDK4的表达,起到控制肿瘤细胞生长及分化的作用,从而干预肺癌转移的形成。Objective To observe the inhibiting effects of Fufei Jian on proliferation and metastasis of lung carcinoma.Methods(1)In vitro:Fufei Jian-containing serum was prepared for culturing lung cancer A549 and SPC-A1 cells,with signal transducer and activator of transcription 3(STAT3)inhibitor AG490 and DDP as positive control drug.Cell count kit 8(CCK-8)assay was used to detect the survival ability of A549 and SPC-A1cells.Scratch assay and Transwell cell invasion assay were used to observe the migration and invasion abilities,respectively.Western blotting assay was used to detect the expression levels of Cyclin D1,cyclin-dependent kinase 4(CDK4),STAT3 and phosphorylated STAT3(p-STAT3).(2)In vivo:Lewis lung cancer model was established in 48 male C57BL/6 mice.After intragastric administration of Fufei Jian and DDP(as positive control drug),respectively,on day 15 all the mice were sacrificed by cervical luxation to observe lung metastatic foci.And then the Xenograft tumors mass was weighed,the inhibiting tumor growth rate and pulmonary metastasis inhibition rate were calculated,the expression levels of Cyclin D1,CDK4 and p-STAT3 were determined by immunohistochemical method.Results The proliferation,migration and invasion abilities of A549 and SPC-A1cells in Fufei Jian group were lower than those in the blank control group,near to the levels of AG490 and DDP treatment groups.In vivo,the inhibiting tumor growth rate in high-dose Fufei Jian group and DDP group was26.67%,52.53%,respectively,and the pulmonary metastasis inhibition rate arrived to 28.76%,33.82%,respectively.The expression levels of p-STAT3,Cyclin D1 and CDK4 in various groups were decreased in vitro and in vivo(P<0.05 or P<0.01).Conclusion Fufei Jian plays a role in controlling tumor cells growth and differentiation through inhibiting activation of STAT3 protein to reduce the expression of Cyclin D1 and CDK4,and thus attenuates metastasis of lung cancer.

关 键 词：扶肺煎 肺癌 侵袭 转移 STAT3信号通路 A549细胞 SPC-A1细胞 Lewis荷瘤小鼠 

分 类 号：R285.5[医药卫生—中药学]