微小RNA-20b对胃癌MGC-803细胞侵袭和迁移的影响  

Effects of microRNA-20b on invasion and migration of gastric cancer MGC-803 cells

作　　者：朱勇[1] 杨哲[1] 李晓花[1] 张云[1] 李剑平 苏云涛[1] ZHU Yong;YANG Zhe;LI Xiaohua;ZHANG Yun;LI Jianping;SU Yuntao(Department of Radiotherapy,Baoji Central Hospital,Baoji 721008,China)

机构地区：[1]宝鸡市中心医院放射治疗科,陕西宝鸡721008 [2]空军军医大学西京医院放射治疗科,710030

出　　处：《临床肿瘤学杂志》2019年第1期21-25,共5页Chinese Clinical Oncology

基　　金：国家自然科学基金青年项目(31500683)

摘　　要：目的探讨微小RNA-20b(miR-20b)对胃癌MGC-803细胞侵袭和迁移的影响。方法采用脂质体Lipo-fectamine 2000法向MGC-803细胞分别转染miR-20b抑制剂(inhibitor转染组)和miR-20b抑制剂阴性对照(NC组),设不行转染的细胞为未转染组;采用实时荧光定量PCR(QPCR)检测各组转染48 h后的miR-20b水平以评估转染效率,分别采用Tran-swell实验和划痕实验比较各组细胞的侵袭和迁移情况,Western blotting检测基质金属蛋白酶-9(MMP-9)、磷酸化信号转导激活转录因子-3(p-STAT3)和磷酸化酪氨酸蛋白激酶-1(p-JAK1)的表达情况。结果 QPCR结果显示,未转染组、NC组和inhib-itor转染组的miR-20b水平分别为1. 054±0. 123、0. 987±0. 085和0. 506±0. 092,与未转染组和NC组相比,inhibitor转染组的miR-20b水平降低(P <0. 05);未转染组、NC组和inhibitor转染组的划痕愈合率分别为(60. 21±1. 16)%、(62. 57±2. 06)%和(25. 17±2. 69)%,穿膜细胞数目分别为(448±39)个、(415±33)个和(196±29)个,与未转染组和NC组相比,inhibitor转染组的划痕愈合率和穿膜细胞数目均降低(P <0. 05)。转染miR-20b抑制剂可降低MMP-9、p-JAK1和p-STAT3的表达水平(P<0. 05)。结论下调miR-20b水平可抑制胃癌细胞侵袭和迁移能力,可能与降低MMP-9表达及抑制JAK/STAT信号通路活性有关,在胃癌靶向治疗中有一定应用前景。Objective To explore the effect of microRNA-20b(miR-20b)on invasion and migration of gastric cancer MGC-803 cells.Methods The MGC-803 cells were transfected with miR-20b inhibitor(inhibitor-transfected group)and miR-20b inhibitor negative control(NC group),and the untransfected cells were assigned as the non-transfected group.Real-time quantitative PCR(QPCR)was used to detect the level of miR-20b at 48 h after transfection in each group to evaluate the transfection efficiency.Transwell and scratch experiments were used to compare the invasive and migrative ability of cells in each group.Western blotting was used to detect the expression of matrix metalloproteinase-9(MMP-9),phosphorylated signal transduction activating transcription factor-3(p-STAT3)and phosphorylated tyrosine protein kinase-1(p-JAK1).Results The results of QPCR showed that the levels of miR-20b in non-transfected group,NC group and inhibitor-transfected group were 1.054±0.123,0.987±0.085 and 0.506±0.092.Compared with non-transfected group and NC group,the level of miR-20b in inhibitor-transfected group were lower(P<0.05).The scratch healing rates in non-transfected group,NC group and inhibitor-transfected group were(60.21±1.16)%,(62.57±2.06)%and(25.17±2.69)%,respectively.The numbers of penetrating cells in non-transfected group,NC group and inhibitor-transfected group were 448±39,415±33 and 196±29,respectively.Compared with non-transfected group and NC group,the scratch healing rates and the number of penetrating cells in inhibitor-transfected group were significantly decreased(P<0.05).Transfection of miR-20b inhibitor significantly reduced the expression of MMP-9,p-JAK1 and p-STAT3(P<0.05).Conclusion Down-regulation of miR-20b level can inhibit the invasion and migration of gastric cancer cells,which may be related to the reduction of MMP-9 expression and the inhibition of JAK/STAT signaling pathway.It has a certain application prospect in targeted therapy of gastric cancer.

关 键 词：胃癌 微小RNA-20b 侵袭迁移 JAK/STAT信号通路 

分 类 号：R735.2[医药卫生—肿瘤]