哈斯也提·艾力,妮拉·马哈德,帕力达·帕拉哈提,崔晓宾.miR-138-5p通过调控Survivin表达对食管鳞癌细胞活性的影响[J].转化医学杂志,2021,10(4):218-223
miR-138-5p通过调控Survivin表达对食管鳞癌细胞活性的影响
The effect of miR-138-5p on the activity of esophageal squamous cell carcinoma cells by regulating the expression of Survivin
  
DOI:
中文关键词:  miR-138-5p  Survivin  食管鳞癌细胞生物活性
英文关键词:miR-138-5p  Survivin  Biological activity of esophageal squamous cell carcinoma cells
基金项目:国家自然科学基金项目(81463062)
作者单位
哈斯也提·艾力 喀什地区第二人民医院 
妮拉·马哈德 喀什地区第二人民医院 
帕力达·帕拉哈提 喀什地区第二人民医院 
崔晓宾 喀什地区第二人民医院 
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中文摘要:
      目的 探究miR-138-5p通过调控Survivin表达对食管鳞癌细胞活性的影响。方法 将实验分为W组(无转染组)、C组(食管鳞癌细胞转染NC组)、Z组(转染miR-138-5p组)。RT-PCR检测Survivin、miR-138-5p水平;Western blot检测Notch1、HIF-1α、MMP-9的表达;Transwell小室测迁移能力;CCK-8检测增殖能力;流式细胞仪检测凋亡情况;双荧光素酶报告基因检测miR-138-5p与Survivin靶向关系。结果 miR-138-5p在食管鳞癌组织中的水平低于癌旁组织(P<0.05);食管鳞癌组织中Survivin水平较癌旁组织高(P<0.05)。miR-138-5p、Survivin在W组与C组中的水平较为接近(P>0.05);与C组相比,Z组miR-138-5p的水平有所上升、Survivin水平有所下降(P<0.05),由此可看出,miR-138-5p转染成功。W组与C组中Notch1、HIF-1α、MMP-9水平较为接近(P>0.05);与C组相比,Z组中Notch1水平有所升高、HIF-1α、MMP-9水平有所下降(P<0.05)。W组食管鳞癌细胞迁移数量(282.67±27.65)与C组细胞迁移数量(279.31±28.22)相差较小(P>0.05);与C组相比,Z组的迁移数量(76.57±6.71)有所降低(P<0.05)。W组与C组在各时间点的OD值较为接近(P>0.05);Z组在各时间点的OD值均较W组与C组低(P<0.05)。W组食管鳞癌细胞凋亡率(2.47±0.11)%与C组食管鳞癌细胞凋亡率(2.45±0.13)%较为接近(P>0.05);Z组食管鳞癌细胞凋亡率(13.32±1.23)%高于W组与C组(P<0.05)。双荧光素梅报告基因检测实验结果显示Survivin′-UTR-WT在miR-NC组表达水平为(0.79±0.09)、miR-138-5p组表达水平为(0.59±0.07),组间比较差异有统计学意义(P<0.001);Survivin′-UTR-MUT在miR-NC组(0.91±0.11)及miR-138-5p组(1.02±0.12)表达无显著差异(P>0.05)。与食管癌组比较,miR-138-5p组大鼠移植瘤的瘤体积、瘤质量降低,抑瘤率升高(P<0.05)。结论 miR-138-5p可通过降低Survivin的表达,抑制食管鳞癌细胞的增殖及迁移,并促进其凋亡。
英文摘要:
      Objective Explore the effect of miR-138-5p on the activity of esophageal squamous cell carcinoma cells by regulating the expression of Survivin. Methods Three groups were set in our study including group W (no transfection group), group C (NC transfection group), and group Z (Transfection of miR-138-5p group). RT-PCR was used to detect the levels of Survivin and miR-138-5p. Western blot was used to detect the expression of Notch1, HIF-1 α, and MMP-9. Transwell chamber was used to detect the migration ability. CCK-8 was used to detect the proliferation ability. Flow cytometry was used to detect apoptosis. The relationship between miR-138-5p and Survivin targeting was detected by dual-luciferase reporter gene. Results The level of Mir-138-5p in esophageal squamous cell carcinoma tissues was lower than that in paracancer tissues (P<0.05). Survivin level in esophageal squamous cell carcinoma tissues was higher than that in adjacent tissues (P<0.05).The levels of Mir-138-5P and Survivin in W group and C group were similar (P>0.05). Compared with group C, the level of Mir-138-5p was increased and the level of Survivin was decreased in group Z (P<0.05), indicating that the transfection of Mir-138-5p was successful. The levels of Notch1, HIF-1α and MMP-9 in group W and GROUP C were similar (P>0.05). Compared with group C, the levels of Notch1 were increased, hiF-1 α and MMP-9 were decreased in group Z (P<0.05).The number of migrating cells in group W was significantly lower than that of in the group W (P< 0.05). The OD values of the group W and group C at each time point were close (P> 0.05), and the OD values of the group Z at each time point were lower than those of in the group W and group C (P< 0.05). The apoptosis rate of ESCC in the group W was (2.47 ± 0.11)%, which was close to that in the group C (2.45 ± 0.13)% (P > 0.05). The apoptosis rate of ESCC in the group Z was (13.32 ± 1.23)%, which was higher than that of in the group W and group C (P< 0.05). The results of double fluorescein plum reporter gene detection showed that the expression level of Survivin '- utr-wt in the mir-nc group was higher (0.79 ± 0.09) than that of in the group and that of in the miR-138-5p group (0.59 ± 0.07, P< 0.001). There was no difference of the expression level of Survivin' - utr-mut in the mir-nc group (0.91 ± 0.11) and miR-138-5p group (1.02 ± 0.12, P > 0.05).Compared with the esophageal cancer group, the tumor volume and tumor mass of rats in Mir-138-5p group were decreased, and the tumor inhibition rate was increased (P<0.05).Conclusion miR-138-5p can inhibit the proliferation and migration of ESCC cells and promote the apoptosis of ESCC cells by reducing the expression of Survivin.
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