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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。
Abstract：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 of PDAC cells was detected by scratch assay and transwell assay. Results The expression of miR-30a-5p in normal pancreatic cell lines was higher than that in PDAC cell lines(P < 0.05). The expression of FAP-1 in normal pancreatic cell lines was lower than that in PDAC cell lines(P < 0.05). MiR-30a-5p and FAP-1 genes had junction sites, and the expression of miR-30a-5p was increased and the expression of FAP-1 significantly decreased after transfection of miR-30a-5p mimic(P < 0.05), and the proliferation and migration ability of PDAC cells decreased(P < 0.05). After transfection with miR-30a-5p inhibitor, the expression of miR-30a-5p was decreased, the expression of FAP-1 significantly increased(P < 0.05), the proliferation and migration of PDAC cells genhanced(P < 0.05). The relationship between FAP-1 and miR-30a-5p and clinicopathologic features of pancreatic cancer patients showed that FAP-1 was related to TNM stage, and FAP-1 related to drinking habits. MiR-30a-5p was correlated with TNM staging(P < 0.05). Conclusion MiR-30a-5p may target the inhibition of FAP-1 protein to inhibit the proliferation and migration of PDAC.
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基本信息:
DOI：10.13885/j.issn.1000-2812.2023.03.002
中图分类号:R735.9
引用信息:
[1]何雨,董保龙,王欣鑫,等.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[J].兰州大学学报(医学版),2023,49(03):5-11.DOI:10.13885/j.issn.1000-2812.2023.03.002.
基金信息:
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）
2023-03-08
2023-03-08