微小RNA-490-3p对人胰腺癌的增殖、迁移、侵袭、凋亡的影响

Rffect of microRNA-490-3p on proliferation,migration,invasion and apoptosis of human pancreatic cell lines AsPC-1 and SW1990

目的观察微小RNA-490-3p(miR-490-3p)对胰腺癌的增殖、迁移、侵袭、凋亡的调控作用。方法选取AsPC-1和SW1990胰腺癌细胞株[均购自美国标准生物品收藏中心(ATCC)细胞库]作为研究对象。将miR-490-3p模拟物(75 pmol)转染至胰腺癌细胞株AsPC-1和SW1990中,分别构建miR-490-3p过表达组,空载体作为阴性对照组,48 h后,利用实时定量聚合酶链反应(Real-time PCR)检测miR-490-3p的表达,细胞计数试剂盒(CCK-8)检测miR-490-3p对细胞增殖的作用,细胞划痕实验检测细胞的迁移能力,Transwell实验检测细胞的侵袭能力,原位缺口末端标记法(TUNEL)实验检测细胞的凋亡能力。此研究时限为4个月。组间数据比较采用独立样本t检验。结果 miR-490-3p在胰腺癌AsPC-1和SW1990细胞中相对表达低于正常胰腺细胞(0.58±0.06、0.40±0.20比1.00±0.11,t=5.886、4.656,P<0.01);转染miR-490-3p模拟物后,胰腺癌AsPC-1和SW1990细胞内miR-490-3p相对表达量明显升高(2 173 099.24±385 195.63、1 842 351.45±498 269.21,t=9.771、6.404,P<0.01),差异有统计学意义。过表达miR-490-3p后,CCK-8实验结果显示,明显降低AsPC-1和SW1990细胞的增殖(1.63±0.06比2.13±0.02,t=3.333,P<0.05;1.09±0.04比1.63±0.03,t=3.119,P<0.05),差异有统计学意义;划痕实验结果显示,明显抑制AsPC-1和SW1990细胞迁移对照组[(48.16±6.17) μm比(18.34±3.26) μm,t=9.564,P<0.01;(48.49±6.98) μm比(23.02±3.26) μm,t=7.390,P<0.01],差异有统计学意义;Transwell细胞侵袭实验显示,明显抑制AsPC-1和SW1990细胞侵袭[(36.00±2.45)个比(15.33±2.94)个,t=13.220,P<0.01;(99.00±15.67)个比(46.17±11.07)个,t=6.745,P<0.01],差异有统计学意义;TUNEL细胞凋亡实验结果显示,(17.75±1.71比6.75±4.03,t=5.025,P<0.01;23.25±3.40比10.25±2.63,t=6.045,P<0.01),差异有统计学意义。结论过表达miR-490-3p后抑制AsPC-1和SW1990细胞株的增殖、迁移、侵袭能力,促进细胞凋亡的作用。

Objective To investigate the effect of microRNA-490-3p(miR-490-3p)on the proliferation,migration,invasion and apoptosis of pancreatic cells.Methods AsPC-1 and SW1990 pancreatic cancer cell lines were selected as the research objects from the ATCC.The miR-490-3p mimic(75 pmol)was transfected into pancreatic cancer cell lines AsPC-1 and SW1990 to construct miR-490-3p overexpression groups,and the empty vector was used as a negative control group.The expression of miR-490-3p was detected by real-time quantiative polymerase chain reaction(Real-time PCR)after 48 h.Cell proliferation was measured by cell counting kit-8(CCK-8)assay.The migration ability was assayed by would healing test.Transwell invasion assay was used to measure cell invasion.The apoptosis ability was assayed by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)test.The study period was 4 months.All data were analyzed using the statistical software SPSS 22.0 for Windows,and the data were presented as mean±SD.The independent-sample t test was used to compare the data between two groups in all pertinent experiments in this study,P<0.05 was considered statistically significant.Results The expression of miR-490-3p in AsPC-1 and SW1990 cells was significantly lower than that in normol pancreatic cells(0.58±0.06,0.40±0.20,t=5.886,4.656,P<0.01).The expression of miR-490-3p in AsPC-1 and SW1990 cells was significantly higher(2173099.24±385195.63 and 1842351.45±498269.21,t=9.771,6.404,P<0.01)after transfection with miR-490-3p mimic.CCK-8 results showed that the proliferation of AsPC-1 and SW1990 cells was significantly reduced(1.63±0.06 vs.2.13±0.02,t=3.333,P<0.05;1.09±0.04 vs.1.63±0.03,t=3.119,P<0.05).The results of wound healing test showed that the migration of AsPC-1 and SW1990 cells was inhibited[(48.16±6.17)μm vs.(18.34±3.26)μm,t=9.564,P<0.01;(48.49±6.98)μm vs.(23.02±3.26)μm,t=7.390,P<0.01].Transwell cell invasion assay showed significant inhibition of AsPC-1 and SW1990 cell invasion[(36.00±2.45)vs.(15.33±2.94),t=13.220,P<0.01;(99.00±15.67)vs.(46.17±11.07),t=6.745,P<0.01].TUNEL cell apoptosis assay showed that the apoptosis rate of AsPC-1 and SW1990 cells was significantly higher[(17.75±1.71)vs.(6.75±4.03),t=5.025,P<0.01;(23.25±3.40)vs.(10.25±2.63),t=6.045,P<0.01].Conclusion Overexpression of miR-490-3p inhibited the proliferation,migration and invasion of AsPC-1 and SW1990 cell lines,and promoted apoptosis.