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

作　　者：哈斯也提·艾力 妮拉·马哈德 帕力达·帕拉哈提 崔晓宾 Hasiyeti·AiLi;NiLa·Mahade;Palida·Palahati;CUI Xiaobin(The Second People's Haspital of kashgar,kashgar Xinjiang 844000,China)

机构地区：[1]喀什地区第二人民医院,新疆喀什844000

出　　处：《转化医学杂志》2021年第4期218-223,共6页Translational Medicine Journal

基　　金：国家自然科学基金项目(81463062)。

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。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-1α,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-1αand MMP-9 in group W and GROUP C were similar(P>0.05).Compared with group C,the levels of Notch1 were increased,hiF-1αand 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 i

关 键 词：miR-138-5p SURVIVIN 食管鳞癌细胞生物活性 

分 类 号：R735.1[医药卫生—肿瘤]