【摘 要】
：
目的 探讨miR-122-3p通过靶向血管内皮生长因子(VEGFA)基因对胰腺癌细胞增殖、克隆、迁移及侵袭能力影响的分子机制.方法 采用实时定量反转录聚合酶链反应(qRT-PCR)检测胰腺癌细胞株(BxPC-3、Capan-1、PANC-1、HPAC、AsPC-1)与正常胰腺导管上皮细胞株HPDE中miR-122-3p mRNA及VEGFA mRNA的表达.将AsPC-1细胞株分为NC组、miR-con组、miR-122-3p mimic组、miR-122-3p+pcDNA组和miR-122-3p+pcD
【机 构】
：
前海人寿广州总医院 肝胆外科,广东 广州 511300;郑州大学附属洛阳中心医院 肝胆外科,河南 洛阳 471000
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目的 探讨miR-122-3p通过靶向血管内皮生长因子(VEGFA)基因对胰腺癌细胞增殖、克隆、迁移及侵袭能力影响的分子机制.方法 采用实时定量反转录聚合酶链反应(qRT-PCR)检测胰腺癌细胞株(BxPC-3、Capan-1、PANC-1、HPAC、AsPC-1)与正常胰腺导管上皮细胞株HPDE中miR-122-3p mRNA及VEGFA mRNA的表达.将AsPC-1细胞株分为NC组、miR-con组、miR-122-3p mimic组、miR-122-3p+pcDNA组和miR-122-3p+pcDNA-VEGFA组.通过噻唑蓝(MTT)细胞增殖实验、平板克隆实验检测各组细胞增殖及克隆能力,通过细胞划痕实验和Transwell实验检测各组细胞迁移和侵袭能力,通过蛋白质印迹实验检测MMP-2、VEGFA、ERK1、MAPK蛋白表达,通过双荧光素酶报告基因法和蛋白质印迹法验证miR-122-3p与VEGFA的靶向关系.结果 各胰腺癌细胞株中miR-122-3p mRNA表达量均低于HPDE细胞株,VEGFA mRNA表达量均高于HPDE细胞株(P<0.05).miR-122-3p mimic组AsPC-1细胞活力、细胞克隆数低于NC组和miR-con组(细胞克隆数20.33±3.31比78.11±8.20、76.49±7.91,t=32.014、28.472,P<0.05),细胞迁移率及穿膜细胞数低于NC组和miR-con组[细胞迁移率(2.16±0.68)％比(13.92±2.46)％、(14.13±2.38)％,t=13.121、12.242,P<0.05;穿膜细胞数(23.67±3.59)个比(91.58±5.00)、(93.01±3.72)个,t=41.716、51.817,P<0.05].miR-122-3p mimic组细胞荧光素酶活性低于NC组和miR-con组(0.51±0.13比2.11±0.24、2.08±0.46,t=17.552、9.279,P<0.05).共转染miR-122-3p和pcDNA-VEGFA后,细胞活力高于miR-122-3p+pcDNA组,细胞迁移率、穿膜细胞数高于miR-122-3p+pcDNA组[细胞迁移率(11.20±1.93)％比(2.35±0.53)％,t=10.609,P<0.05;穿模细胞数(84.39±2.92)个比(22.15±2.50)个,t=49.435,P<0.05],细胞内MMP-2、VEGFA、ERK1、MAPK蛋白表达水平升高(P<0.05).结论 miR-122-3p在胰腺癌细胞中低表达,过表达miR-122-3p可抑制胰腺癌细胞恶性生物学行为,机制可能与靶向VEGFA有关.
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