小白菊内酯通过调控微小RNA-637表达抑制瘢痕疙瘩成纤维细胞增殖、迁移和侵袭的研究

Parthenolide inhibits the proliferation,migration and invasion of keloid fibroblasts by regulating the expression of microRNA-637

目的研究小白菊内酯是否通过调控微小RNA-637(miR-637)表达抑制瘢痕疙瘩成纤维细胞增殖、迁移和侵袭。方法分离培养瘢痕疙瘩成纤维细胞和正常皮肤成纤维细胞。瘢痕疙瘩成纤维细胞分为对照组(未处理)、低、中、高剂量实验组(5,10,20μmol·L^(-1)小白菊内酯)、miR-NC组(转染miR-NC)、miR-637组(转染miR-637 mimic)、PTL+anti-miR-NC组(20μmol·L^(-1)小白菊内酯+转染anti-miR-NC)、PTL+anti-miR-637组(20μmol·L^(-1)小白菊内酯+转染miR-637 inhibitor)。用噻唑蓝法检测细胞的增殖情况,用Transwell法检测细胞的迁移和侵袭情况,用实时定量聚合酶链反应检测miR-637的表达情况。结果低、高剂量实验组和对照组、miR-NC组、miR-637组、PTL+anti-miR-NC和PTL+anti-miR-637组的增殖能力(光密度值)分别为0.71±0.04,0.39±0.03,0.84±0.05,0.82±0.07,0.44±0.03,0.38±0.03和0.73±0.05,迁移细胞数分别为(93.75±7.35),(58.18±3.57),(124.31±9.87),(125.05±8.08),(63.15±5.04),(57.57±3.23)和(103.79±6.68)个,侵袭细胞数分别为(81.60±6.16),(42.40±3.09),(105.75±7.80),(106.41±9.96),(52.82±4.19),(40.27±4.11)和(90.51±6.96)个,miR-637表达水平分别为1.78±0.15,3.36±0.28,1.00±0.05,1.00±0.07,2.77±0.20,1.00±0.05和0.29±0.03。低、高剂量实验组的上述指标与对照组比较,miR-637组的上述指标与miR-NC组比较,PTL+anti-miR-637组的上述指标与PTL+anti-miR-NC组比较,差异均有统计学意义(均P<0.05)。正常皮肤成纤维细胞和瘢痕疙瘩成纤维细胞中miR-637表达水平分别为1.00±0.08和0.45±0.04,差异有统计学意义(P<0.05)。结论小白菊内酯通过增加miR-637的表达水平,抑制瘢痕疙瘩成纤维细胞增殖、迁移和侵袭。

Objective To investigate whether parthenolide inhibits the proliferation,migration and invasion of keloid fibroblasts by regulating the expression of microRNA-637(miR-637).Methods Keloid fibroblasts and normal skin fibroblasts were separated and cultured.Keloid fibroblasts were divided into control group(untreated),experimental-L,-M,-H groups(5,10,20μmol·L^(-1) parthenolide),miR-NC group(transfected with miR-NC),miR-637 group(transfected with miR-637 mimic),PTL+anti-miR-NC group(20μmol·L^(-1) parthenolide+transfected with anti-miR-NC)and PTL+anti-miR-637 group(20μmol·L^(-1)parthenolide+transfection miR-637 inhibitor).The thiazole blue method was used to detect cell proliferation.The Transwell method was applied to monitor cell migration and invasion.The real-time quantitative polymerase chain reaction was employed to determine the expression of miR-637.Results The proliferation abilities(optical density value)of experimental-L,-H groups and control,miR-NC,miR-637,PTL+anti-miR-NC and PTL+anti-miR-637 groups of keloid fibroblasts were 0.71±0.04,0.39±0.03,0.84±0.05,0.82±0.07,0.44±0.03,0.38±0.03 and 0.73±0.05;the number of migrating cells was 93.75±7.35,58.18±3.57,124.31±9.87,125.05±8.08,63.15±5.04,57.57±3.23 and 103.79±6.68;the number of invasive cells was 81.60±6.16,42.40±3.09,105.75±7.80,106.41±9.96,52.82±4.19,40.27±4.11 and 90.51±6.96;the expression levels of miR-637 were 1.78±0.15,3.36±0.28,1.00±0.05,1.00±0.07,2.77±0.20,1.00±0.05 and 0.29±0.03,respectively.There were significant differences in the above indexes between experimental-L,-H groups and control group,between miR-637 group and miR-NC group,and between PTL+anti-miR-637 group and PTL+anti-miR-NC group(all P<0.05).The expression levels of miR-637 in normal skin fibroblasts and keloid fibroblasts were 1.00±0.08 and 0.45±0.04,and the difference was statistically significant(P<0.05).Conclusion Parthenolide can inhibit the proliferation,migration and invasion of keloid fibroblasts by increasing the level of miR-637.