法尼醇受体通过下调miR-21水平抑制非小细胞肺癌细胞增殖及侵袭能力  

Farnesoid X receptor inhibits the proliferation and invasion of non-small cell lung cancer cells by downregulating miR-21

作　　者：王振华[1] 陈金亮 邢媛媛 郭喜喜[1] 李东飞 WANG Zhenhua;CHEN Jinliang;XING Yuanyuan;GUO Xixi;LI Dongfei(the Second Department of Thoracic Oncology,Xinxiang Central Hospital,Xinxiang 453000,China;Department of Respiratory Medicine,Nantong First Hospital,Nantong 226006)

机构地区：[1]新乡市中心医院胸瘤二科,河南新乡453000 [2]南通市第一医院呼吸科,江苏南通226006

出　　处：《中国比较医学杂志》2021年第9期43-50,共8页Chinese Journal of Comparative Medicine

基　　金：江苏省科技项目(BK20191207)。

摘　　要：目的观察法尼醇受体(FXR)在非小细胞肺癌(NSCLC)组织和细胞系中的表达水平,探讨了FXR在NSCLC细胞增殖和侵袭中的作用是否与调控miR-21水平相关。方法在2010年至2012年间,取本院手术切除的80例NSCLC组织及配对的正常组织。通过免疫组化或Western blot分析NSCLC组织和细胞系中FXR表达。以NSCLC细胞A549为研究对象,通过CCK-8法、Transwell法和qRT-PCR法评估不同浓度的GW3965(1~5μmol/L)对细胞活力、侵袭和miR-21水平影响。通过转染技术来过表达或敲低miR-21表达,并评估其对细胞活力、侵袭的影响。采用双荧光素酶报告分析FXR与miR-21之间的关系。结果与邻近的非肿瘤正常组织相比,FXR在NSCLC组织中的表达显著降低(P<0.001)。Kaplan-Meier分析显示FXR低表达预示NSCLC患者预后较差(χ~2=4.496,P=0.033)。FXR高、低表达的NSCLC在临床分期、肿瘤大小、T分期和淋巴结转移(N分期)上有显著性差异(P<0.05)。GW3965以剂量依赖方式促进了A549细胞中FXR蛋白表达,抑制了细胞的增殖和侵袭,并降低了miR-21表达。miR-21过表达显著逆转了FXR对NSCLC细胞生长和侵袭的抑制作用(P<0.05),并且miR-21沉默显著增强FXR对NSCLC细胞生长和侵袭的抑制作用(P<0.05)。双荧光素酶报告证实FXR通过靶向miR-21来抑制NSCLC细胞增殖和侵袭。Spearman分析显示,NSCLC标本中FXR与miR-21的表达呈强负相关(P<0.001)。KaplanMeier分析显示,"FXR低表达"和"miR-21高表达"共存模式预测了NSCLC患者最差的预后(χ~2=8.201,P=0.004)。结论 FXR通过下调miR-21抑制NSCLC细胞的生长,提示调控FXR/miR-21可能是治疗NSCLC的潜在策略。Objective To observe the expression levels of Farnesoid X receptor(FXR)in non-small cell lung cancer(NSCLC)tissue and cell lines,and to explore whether the role of FXR in the proliferation and invasion of NSCLC cells is related to the regulation of miR-21.Methods From 2010 to 2012,80 cases of NSCLC tissue and matched normal tissue were collected.The expression of FXR in the NSCLC tissue and cell lines was analyzed using immunohistochemistry or Western blot.The effects of GW3965(1~5μmol/L)on cell viability and invasion and miR-21 levels were evaluated using CCK-8,transwell and qRT-PCR.MiR-21 was overexpressed or knocked down using transfection technology,and the effects on cell viability and invasion were evaluated.Moreover,the relationship between FXR and miR-21 was analyzed by double luciferase assay.Results Compared with the adjacent non-tumor normal tissue,the expression of FXR was significantly lower in NSCLC tissue(P<0.001).Kaplan-Meier analysis revealed that low FXR expression predicted a worse prognosis for NSCLC patients(χ2=4.496,P=0.033).There were significant differences in clinical stage,tumor size,T stage and lymph node metastasis(N stage)between high-and low-FXR tumors(P<0.05).GW3965 promoted FXR protein expression in A549 cells in a dose-dependent manner,inhibited cell proliferation and invasion and decreased miR-21 expression.Overexpression of miR-21 significantly reversed the inhibitory effects of FXR on the growth and invasion of NSCLC cells(P<0.05),and miR-21 silencing significantly enhanced the inhibitory effects of FXR on the growth and invasion of NSCLC cells(P<0.05).The double luciferase assay confirmed that FXR inhibited the proliferation and invasion of NSCLC cells by targeting miR-21.Spearman’s test revealed a strong negative correlation between FXR and miR-21 expression in NSCLC specimens(P<0.001).Furthermore,Kaplan-Meier analysis revealed that a coexistence pattern of"FXR low"and"miR-21 high"predicted the worst prognosis in NSCLC patients(χ2=8.201,P=0.004).Conclusions FXR inhibits

关 键 词：法尼醇受体 MIR-21 非小细胞肺癌 增殖 侵袭 

分 类 号：R-33[医药卫生]