[目的]探索miR-217与NRP1在奥沙利铂耐药性胃癌细胞中的作用机制。[方法]将MGC803/L-OHP细胞分为4组:miR NC组、miR-217 mimic组、si NC组与si NRP1组。通过实时荧光定量PCR实验和蛋白免疫印迹实验检测耐药和非耐药细胞中miR-217和NRP1的表达水平;通过MTT实验分析MGC803/L-OHP细胞的增殖能力;通过Transwell实验分析MGC803/L-OHP细胞的转移活性;通过流式细胞术分析MGC803/L-OHP细胞的凋亡率;通过荧光素酶报告基因实验分析MGC803/L-OHP细胞中miR-217与NRP1的靶向关系。[结果]上调miR-217或抑制NRP1能够抑制非耐药MGC803细胞生长。与MGC803细胞比较，MGC803/L-OHP细胞中的miR-217表达水平降低(0。79±0。08 vs 0。21±0。02;P＜0。05)，NRP1 表达水平增加(0。41±0。09 vs 0。78±0。12;P＜0。05)。与 miR NC 组比较，miR-217 mimic组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(2。37±0。23 vs 25。38±5。12;P＜0。05)。与si NC组比较，si NRP1组的MGC803/L-OHP细胞的增殖活性降低、侵袭能力降低、凋亡率增加(4。67±0。83 vs 26。17±7。19;P＜0。05)。荧光素酶活性测定显示，和miR NC组比较，在NRP1-WT组中转染miR-217 mimic后荧光素酶活性显著降低(0。87±0。03 vs 0。26±0。02;P＜0。05)。[结论]在奥沙利铂耐药性胃癌细胞中，miR-217低表达而NRP1高表达，并且miR-217能够靶向抑制NRP1的表达。通过上调miR-217或抑制NRP1的表达能够减弱胃癌细胞对奥沙利铂的耐药性。
Effect of miR-217 on oxaliplatin resistance of gastric cancer cells by regulating NRP1
[Objective]To explore the mechanism of miR-217 and NRP1 in oxaliplatin-resistant gastric cancer cells.[Method]MGC803/L-OHP cells were divided into four groups:miR NC group,miR-217 mimic group,si NC group and si NRP1 group.The expression levels of miR-217 and NRP1 in drug-resistant and non-drug-resistant cells were detected by real-time fluorescent quantitative PCR and Western Blot.The proliferation ability of MGC803/L-OHP cells was analyzed by MTT assay.Transwell assay was used to analyze the metastatic activity of MGC803/L-OHP cells.The apoptosis rate of MGC803/L-OHP cells was analyzed by flow cytometry.Luciferase reporter assay was used to analyze the targeting relationship between miR-217 and NRP1 in MGC803/L-OHP cells.[Result]Up-regulation of miR-217 or inhibition of NRP1 could inhibit the growth of non-drug resistant MGC803 cells.Compared with MGC803 cells,the expression level of miR-217 in MGC803/L-OHP cells was decreased(0.79±0.08 vs 0.21±0.02;P＜0.05)and increased NRP1 expression(0.41±0.09 vs 0.78±0.12;P＜0.05).Compared with the miR NC group,the proliferation activity and invasion ability of MGC803/L-OHP cells in the miR-217 mimic group were decreased,and the apoptosis rate was increased(2.37±0.23 vs 25.38±5.12;P＜0.05).Compared with the si NC group,the proliferation activity and invasion ability of MGC803/L-OHP cells in the si NRP1 group were decreased,and the apoptosis rate was increased(4.67±0.83 vs 26.17±7.19;P＜0.05).Luciferase activity assay showed that compared with miR NC group,the luciferase activity was significantly decreased in NRP1-WT group after transfection with miR-217 mimic(0.87±0.03 vs 0.26±0.02;P＜0.05).[Conclusion]miR-217 is down-regulated and NRP1 is up-regulated in oxaliplatin-resistant gastric cancer cells,and miR-217 can target and inhibit the expression of NRP1.The oxaliplatin resistance of gastric cancer cells can be inhibited by up-regulation of miR-217 or inhibition of NRP1 expression.
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