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目的探讨微小RNA分子miR-214对食管鳞癌细胞Eca109侵袭能力的影响及其可能的分子机制。方法根据人源miR-214序列合成双链模拟物。通过脂质体转染将miR-214模拟物分子导入食管鳞癌Eca109细胞中,转染无关miRNA模拟物作为对照。采用定量PCR法检测细胞中成熟miR-214分子水平,以Matrigel包被的Transwell实验测定侵袭细胞率,通过蛋白质印迹法检测细胞中E-钙黏蛋白(E-cadherin)的表达,利用流式细胞术检测E-cadherin阳性细胞率。结果 Eca109细胞转染miR-214模拟物48h后,其成熟miR-214水平较对照组明显上升(P<0.01);细胞侵袭能力较对照组降低(P<0.05)。同时,miR-214转染组的E-cadherin表达水平及E-cadherin阳性细胞率较对照组下降(P<0.05)。结论 miR-214可能通过抑制细胞上皮间质转化(EMT)的方式抑制食管鳞癌细胞的侵袭能力。
Objective To investigate the effect of microRNA miR-214 on the invasion of esophageal squamous cell carcinoma Eca109 and its possible molecular mechanism. Methods Double-stranded mimetics were synthesized based on human miR-214 sequences. The miR-214 mimic molecule was introduced into esophageal squamous cell carcinoma Eca109 cells by lipofection and transfected with an irrelevant miRNA mimic as a control. Quantitative PCR was used to detect the level of mature miR-214 cells. Matrigel-coated Transwell assay was used to determine the rate of invasive cells. Western blotting was used to detect the expression of E-cadherin in cells and flow cytometry was used. The rate of E-cadherin positive cells was measured. Results After miR-214 mimics were transfected into Eca109 cells for 48h, the mature miR-214 levels were significantly higher than those in the control group (P<0.01). The invasiveness of miR-214 cells was lower than that of the control group (P<0.05). At the same time, the expression of E-cadherin and E-cadherin positive cells in the miR-214 transfection group was lower than that in the control group (P<0.05). Conclusion miR-214 may inhibit the invasion of esophageal squamous carcinoma cells by inhibiting epithelial-mesenchymal transition (EMT).