长链非编码RNA FEZF1-AS1对胃癌细胞株SGC-7901增殖和侵袭的影响

Effects of long intergenic non - coding RNA FEZF1 - AS1 on proliferation and invasion of gastric cancer cells SGC -7901

目的观察长链非编码RNAFEZF1-ASl（LineFEZF1-ASl）的表达对胃癌细胞株SGC-7901增殖及侵袭的影响。方法胃癌SGC-7901细胞分为两组：FEZF1-ASl小干扰RNA（sir-NA）组（si-FEZF1-ASI）和阴性对照组（si．NC）。通过siRNA干扰SGC-7901细胞中LineFEZF1-ASl的表达，采用克隆形成实验、细胞计数试剂盒（CCK．8）、划痕和Transwell侵袭实验检测胃癌细胞增殖及侵袭能力的变化，应用实时定量聚合酶链反应（Real-timePCR）和Westernblot检测上皮-间充质转化（EMT）相关标志物的表达水平。结果克隆形成实验结果显示，与si-NC组比较，si-FEZF1-ASl组细胞的克隆形成能力受到抑制[（43．22±4．35）％比（26．78±3．34）％，t=5．194，P：0．007]；CCK-8实验显示，3d后si-FEZF1-ASI组细胞的抑制率明显高于si-NC组[（31．32±4．48）％比（13．86±0．64）％，k11．590，P：0．000]；划痕试验结果显示，si-FEZF1-ASl组痕间问距明显大于si-NC组[（598．67±18．81）μm比（428．32±13．90）μm，t=12．620，P=0．000]；Transwell结果显示，si-FEZF1-ASl组侵袭转移细胞数明显减少[（60．50±5．13）个比（103．50±9．54）个，t=7．942，P=0．001]；liT-PCR结果表明，si-FEZF1-ASl组E-钙黏蛋白（E．cadherin）mtlNA的相对表达水平明显上升[（2．01±0．36），t=5．435，P=0．002]；波形蛋白（Vimentin）、p-连环蛋白（p-catenin）mRNA的相对表达水平明显下降[（0．54±0．05），t；9．734，P=0．000；（0．50±0．10），t=8．019，P=0．000]；Westernblot实验结果显示，E．cadherin蛋白水平，si．FEZF1，ASl组较si-NC组明显上升[（86304±447）比（35623±227），t=175．100，P；0．000]；Vimentin、B-catenin蛋白水平，si-FEZF1-AS1组较si-NC组明显降低[（29135±89）比（82253±353），t=252．800，P=0．000；（15283±147）比（42506±245），t=165．200，P=0。000]。结论干扰LineFEZF1-ASl后可抑制胃癌细胞SGC-7901的增殖和侵袭能力，其机制可能与Wnt／β-eatenin信号通路有关。

Objective To investigate the effects of long intergenic non - coding RNA （ Linc ） FEZF1 -AS1 on proliferation and invasion of SGC -7901 cells. Methods The SGC -7901 cells were classified into two groups ： FEZF1 - AS1 small interfering RNA （ si - FEZF1 - AS1 ） and the negative control siRNA （ si - NC）. Linc FEZF1 - ASI was knocked down in si - FEZF1 - AS1 group by small interfering RNA （siRNA）. The proliferative activity of SGC -7901 cells was determined by colony formation assay and cell counting kit - 8 （ CCK - 8） assay, and the invasion ability was studied by wound healing assay and Transwell assay. The expression levels of epithelial - mesenchymal transition （EMT） - related molecules was detected by RT - PCR and Western blotting. Results The results of colony formation assay showed that the colony - forming capacity was impaired in si - FEZF1 - AS1 group compared to the si - NC group [ （26. 78 ±3.34）% vs. （43.22±4. 35）% ,t =5. 194,P =0. 007] ; CCK -8 results revealed that the proliferation of SGC - 7901 cells was dramatically inhibited in si - FEZF1 - AS1 group after 3 days [ （31.32 ±4.48）% vs. （13.86 ±0. 64）% ,t = 11. 590,P =0. 000] ; Scratch tests showed that, 48 h after scratches in SGC -7901 cells, the space between scratches was significantly increased as compared withthat insi-NCgroup E（598-67 ±18.81） vs. （428.32±13.90） Ixm,t=12.620,P=0.000]; The results of Transwell assay also indicated that the number of metastatie cells in the si - FEZF1 - AS1 group was significantly decreased compared to the si - NC group [ （ 60. 50 ± 5.13 ） vs. （ 103.50 ± 9. 54 ）, t = 7. 942 ,P =0. 001 ] ; The results of RT - PCR showed that the relative mRNA expression levels of E - eadherin were increased in si - FEZF1 - AS1 group E （2. 01 ±0.36）, t = 5. 435,P = 0. 002 ] , and those of Vi- mentin and 13 - eatenin were decreased [ （0.54 ± 0. 05 ） , t = 9. 734, P = 0. 000 ; （0.50 ± 0. 10 ）, t = 8. 019 ,P =0. 000]. The results of Western blotting revealed that the expression levels of E - eadherin in si - FEZF1 - AS1 group were significantly increased as compared with the si - NC group [ （ 86 304 ± 447 ） vs. （35623 ± 227）, t= 175.100, P= 0.000], and those of Vimentin and 13 - eatenin in the si - FEZF1 - AS1 group were signifieantly decreased as compared with the si - NC group [ （ 29 135 ± 89 ） vs. （82253 ± 353）,t=252.800,P=0.000; （15283 ± 147） vs. （42506 ± 245）,t=165.200,P= 0. 000 ]. Conclusion Dysregulation of Line FEZF1 - AS1 can suppress proliferation and invasion of SGC - 7901 cells, which may be related to the Wnt/B - eatenin siznaling oathwav.