MiR-30a-5p靶向成纤维蛋白-1抑制胰腺导管腺癌增殖和迁移作用  

MiR-30a-5p targeting fibrinogen-1 inhibits proliferation and migration of pancreatic ductal adenocarcinoma application

作　　者：何雨 董保龙 王欣鑫 李渊[1] 杨晓军[1,2,3,4] He Yu;Dong Baolong;Wang Xinxin;Li Yuan;Yang Xiaojun(Department of General Surgery,Gansu Provincial Hospital,Lanzhou 730000,China;Gansu Key Laboratory of Molecular Diagnostics and Precision Medicine for Surgical Oncology,Gansu Provincial Hospital,Lanzhou 730000,China;Gansu Engineering Research Center for Prevention and Control of Gastrointestinal Malignant Tumors,Gansu Provincial Hospital,Lanzhou 730000,China;The First Clinical Medical School of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]甘肃省人民医院普外科,甘肃兰州730000 [2]甘肃省人民医院甘肃省外科肿瘤分子诊断与精准治疗重点实验室,甘肃兰州730000 [3]甘肃省人民医院甘肃省消化道恶性肿瘤防控工程研究中心,甘肃兰州730000 [4]兰州大学第一临床医学院,甘肃兰州730000

出　　处：《兰州大学学报（医学版）》2023年第3期5-11,共7页Journal of Lanzhou University（Medical Sciences）

基　　金：国家级科研培育计划院内重点资助项目(2019-216);甘肃省创新基地和人才资助项目(20JR10RA433);甘肃省科技计划重点研发资助项目(21YF5WA027);甘肃省卫生健康行业科研计划资助项目(GSWSKY2020-45)。

摘　　要：目的 探讨microRNA-30a-5p (miR-30a-5p)对胰腺导管腺癌(PDAC)细胞增殖和迁移的影响及对成纤维细胞活化蛋白-1 (FAP-1)的调控作用。方法 反转录聚合酶链反应、免疫印迹法分别检测PDAC细胞及胰腺正常细胞中miR-30a-5p和FAP-1的表达。生物信息学网站预测miR-30a-5p、FAP-1基因存在结合位点,FAP-1在正常细胞与癌细胞表达存在差异;miR-30a-5p模拟物、抑制物,阴性对照物转染PDAC细胞系,反转录聚合酶链反应检测转染效率,蛋白质印迹法检测FAP-1的蛋白表达水平;CCK-8法检测PDAC细胞的增殖;划痕实验、细胞迁移实验检测PDAC细胞的迁移作用。结果 MiR-30a-5p在胰腺正常细胞系中的表达高于PDAC细胞系(P<0.05);FAP-1在胰腺正常细胞系中的表达低于PDAC细胞系(P <0.05)。MiR-30a-5p、FAP-1基因有连接位点,转染miR-30a-5p模拟物后miR-30a-5p表达量增高,FAP-1的表达量显著降低(P <0.05),PDAC细胞的增殖和迁移能力下降(P <0.05);转染miR-30a-5p抑制物后miR-30a-5p表达量降低,FAP-1的表达量显著增加(P <0.05),PDAC细胞增殖和迁移能力增强(P <0.05)。FAP-1、miR-30a-5p与胰腺癌患者临床病理特征之间的关系表明:FAP-1与肿瘤TNM分期相关,FAP-1与饮酒习惯相关;miR-30a-5p与肿瘤TNM分期相关(P <0.05)。结论 MiR-30a-5p可能靶向抑制FAP-1蛋白,抑制PDAC增殖、迁移。Objective To investigate the effect of miR-30a-5p on pancreatic ductal adenocarcinoma(PDAC)cell proliferation and migration and its regulatory effect on fibroblast-1(FAP-1).Method The expression of miR-30a-5p and FAP-1 in PDAC cell line and normal pancreatic cell line was detected by Western blotting and reverse transcription PCR(RT-PCR).The online bioinformatics website predicted that the expression difference of FAP-1 gene in normal cells and PDAC cell lines,and there were binding sites of miR-30a-5p and FAP-1 gene.PDAC cell lines were transfected with miR-30a-5p mimic,inhibitor and negative control sequence.The transfection efficiency was detected by RT-PCR,and the protein expression level of FAP-1 was detected by Western blotting. The proliferation of PDAC cells was detected by CCK-8 assay. The migration ofPDAC cells was detected by scratch assay and transwell assay. Results The expression of miR-30a-5p innormal pancreatic cell lines was higher than that in PDAC cell lines (P < 0.05). The expression of FAP-1 innormal pancreatic cell lines was lower than that in PDAC cell lines (P < 0.05). MiR-30a-5p and FAP-1 geneshad junction sites, and the expression of miR-30a-5p was increased and the expression of FAP-1 significantlydecreased after transfection of miR-30a-5p mimic (P < 0.05), and the proliferation and migration ability ofPDAC cells decreased (P < 0.05). After transfection with miR-30a-5p inhibitor, the expression of miR-30a-5pwas decreased, the expression of FAP-1 significantly increased (P < 0.05), the proliferation and migration ofPDAC cells genhanced (P < 0.05). The relationship between FAP-1 and miR-30a-5p and clinicopathologicfeatures of pancreatic cancer patients showed that FAP-1 was related to TNM stage, and FAP-1 related todrinking habits. MiR-30a-5p was correlated with TNM staging (P< 0.05). Conclusion MiR-30a-5p may targetthe inhibition of FAP-1 protein to inhibit the proliferation and migration of PDAC.

关 键 词：肿瘤微环境 胰腺导管腺癌 成纤维细胞活化蛋白-1 

分 类 号：R735.9[医药卫生—肿瘤]