健脾消癌方通过lncRNA HOTAIR/JAK2/STAT3信号通路抑制结肠癌细胞株HCT116转移的机制  

Efficacy of Jianpi Xiaoai Prescription in Inhibition of Metastasis of Colon Cancer HCT116Cells:An Exploration Based on LncRNA HOTAIR/JAK2/STAT3 Signaling Pathway

作　　者：焦蕉 唐麒[1] 蒋益兰 李东芳[2] 刘佳琴[3] 胡广生 JIAO Jiao;TANGQi;JIANG Yi-lan;LI Dong-fang;LIU Jia-qin;HU Guang-sheng(Graduate School,Hunan University of Chinese Medicine,Changsha 410006,China;Hunan Cancer Hospital,Changsha 410013,China;Hunan Academy of Traditional Chinese Medicine Affiliated Hospital,Changsha 410208,China)

机构地区：[1]湖南中医药大学研究生院,长沙410208 [2]湖南省肿瘤医院,长沙410013 [3]湖南省中医药研究院附属医院,长沙410006

出　　处：《中国实验方剂学杂志》2021年第23期66-71,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81774287);黎月恒全国名老中医药专家传承工作室项目(湘财社【2014】119号);湖南省中医药管理局青年课题项目(2021193);长沙市科技局一般课题项目(kq1706040);湖南创新型省份建设专项(2019SK4013)。

摘　　要：目的:观察健脾消癌方对Hox转录本反义基因间RNA(lncRNA HOTAIR)/酪氨酸蛋白激酶2(JAK2)/信号传导及转录激活因子3(STAT3)信号通路的影响,探讨健脾消癌方抑制结肠癌转移的可能作用机制。方法:分析不同细胞株lncRNA HOTAIR的表达情况,采用10%,15%,20%健脾消癌方含药血清作用于HCT116细胞,transwell实验检测健脾消癌方对HCT116细胞的侵袭能力影响,实时荧光定量聚合酶链式反应(Real-time PCR)检测lncRNA HOTAIR,JAK2,STAT3 mRNA的表达,蛋白免疫印迹法(Western blot)检测JAK2,磷酸化STAT3(p-STAT3),STAT3蛋白表达情况。结果:结肠癌细胞株HCT116中lncRNA HOTAIR的表达水平最高,采用此细胞作为观察对象。与空白组比较,健脾消癌方各组的侵袭细胞数均降低(P<0.05,P<0.01);健脾消癌方中剂量组JAK2 mRNA相对表达水平降低(P<0.05);健脾消癌方中、高剂量组的lncRNA HOTAIR,健脾消癌方高剂量组的JAK2 mRNA相对表达水平显著降低(P<0.01)。与空白组比较,健脾消癌方中剂量组的p-STAT3蛋白相对表达水平均降低(P<0.05);健脾消癌方中、高剂量组的JAK2蛋白,健脾消癌方高剂量组的p-STAT3蛋白相对表达水平均明显降低(P<0.01)。结论:健脾消癌方可有效抑制结肠癌细胞转移,其机制可能与抑制lncRNA HOTAIR/JAK2/STAT3信号通路的表达相关。Objective:To observe the effect of Jianpi Xiaoai prescription on long non-coding RNA Hox transcript antisense intergenic RNA(lncRNA HOTAIR)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway and to explore the potential mechanism of Jianpi Xiaoa prescription in suppressing the metastasis of colon cancer.Method:The expression of lncRNA HOTAIR in different cells was analyzed.Following the treatment of HCT116 cells with 10%,15%,and 20%Jianpi Xiaoai prescription-containing serum,the invasive ability of Jianpi Xiaoai prescription on HCT116 cells was assessed by transwell assay.The mRNA expression levels of lncRNA HOTAIR,JAK2,and STAT3 were measured by Realtime polymerase chain reaction(Real-time PCR),and the protein expression levels of JAK2,phosphorylated STAT3(p-STAT3)and STAT3 by Western blot.Result:The highest expression of lncRNA HOTAIR was detected in HCT116 cells.Compared with the blank group,each Jianpi Xiaoai prescription group exhibited a decreased number of invasive cells(P<0.05,P<0.01).The relative JAK2 mRNA expression in the middle-dose Jianpi Xiaoai prescription group was down-regulated(P<0.05),and the relative lncRNA HOTAIR mRNA expression in the middle-and high-dose Jianpi Xiaoai prescription groups and the relative JAK2 mRNA expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (P<0.01).Compared with the blank group,the relative p-STAT3 protein expression was down-regulated in the middle-dose Jianpi Xiaoai prescription group(P<0.05),and the relative JAK2 protein expression in the middle-and highdose Jianpi Xiaoai prescription groups and the relative p-STAT3 protein expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (P<0.01).Conclusion:Jianpi Xiaoai prescription effectively inhibits the metastasis of colon cancer cells,which may be related to the inhibition of lncRNA HOTAIR/JAK2/STAT3 signaling pathway.

关 键 词：健脾消癌方 结肠癌 HCT116细胞 Hox转录本反义基因间RNA 酪氨酸蛋白激酶2 信号传导及转录激活因子3 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]