LncRNA SOX2-OT调节miR-194-5p/SRXN1轴对食管鳞状细胞癌细胞发生发展的影响OA
Effect of LncRNA SOX2-OT on the Occurrence and Development of Esophageal Squamous Cell Carcinoma Cells by Regulating the miR-194-5p/SRXN1 Axis
目的 探讨长链非编码RNA(LncRNA)性别决定相关基因簇2重叠转录本(SOX2-OT)调节微小RNA-194-5p(miR-194-5p)/硫氧还蛋白-1(SRXN1)轴对食管鳞状细胞癌(ESCC)细胞发生发展的影响.方法 以人ESCC细胞TE-1细胞为研究对象,将其分为Control组(正常培养),sh-NC组(转染sh-NC)、sh-SOX2-OT组(转染sh-SOX2-OT)、sh-SOX2-OT+anti-NC 组(转染 sh-SOX2-OT+anti-NC)、sh-SOX2-OT+anti-miR-194-5p 组(转染 sh-SOX2-OT+anti-miR-194-5p).qRT-PCR 法检测各组 TE-1 细胞中 LncRNA SOX2-OT、miR-194-5p、SRXN1 mRNA 表达;EdU 染色、Transwell 实验、流式细胞仪分别检测各组TE-1细胞的增殖、迁移和侵袭、凋亡;Western Blot检测TE-1细胞中PCNA、SRXN1、MMP-9、Cleaved Caspase-3蛋白的表达;双荧光素酶报告基因实验检测LncRNA SOX2-OT与miR-194-5p、miR-194-5p与SRXN1的相互作用.结果 sh-SOX2-OT组EdU阳性细胞率、迁移数、侵袭数、LncRNA SOX2-OT、SRXN1 mRNA和PCNA、SRXN1、MMP-9 蛋白低于 sh-NC 组、Control 组,凋亡率、miR-194-5 p、Cleaved Caspase-3 蛋白表达高于 sh-NC 组、Control 组(P＜0.05);与 sh-SOX2-OT 组、sh-SOX2-OT+anti-NC 组相比,sh-SOX2-OT+anti-miR-194-5p 组 EdU 阳性细胞率、迁移数、侵袭数、SRXN1 mRNA和蛋白、PCNA、SRXN1、MMP-9蛋白表达升高,凋亡率、miR-194-5p、Cleaved Caspase-3蛋白表达降低(P＜0.05).LncRNA SOX2-OT可以靶向负调控miR-194-5p;miR-194-5p可以靶向负调控SRXN1.结论 敲除LncRNA SOX2-OT可抑制ESCC细胞的发生发展,其机制可能是通过调节miR-194-5p/SRXN1信号通路实现的.
Objective To investigate effects of long non coding RNA(LncRNA)sex determining region Y-box 2 overlap-ping transcript(SOX2-OT)on the occurrence and development of esophageal squamous cell carcinoma(ESCC)cells by regula-ting the microRNA-194-5p(miR-194-5p)/thioredoxin 1(SRXN1)axis.Methods Human ESCC TE-1 cells were grouped into Control group(normal culture),sh-NC group(transfected with sh-NC),sh-SOX2-OT group(transfected with sh-SOX2-OT),sh-SOX2-OT+anti-NC group(transfected with sh-SOX2-OT+anti-NC),and sh-SOX2-OT+anti-miR-194-5p group(transfected with sh-SOX2-OT+anti-miR-194-5p).QRT-PCR method was applied to detect mRNA expression of LncRNA SOX2-OT,miR-194-5p,and SRXN1 in various TE-1 cells.EdU staining,Transwell assay,and flow cytometry were used to measure the prolifera-tion,migration,invasion,and apoptosis of TE-1 cells in each group.Western Blot was applied to detect the expression of PCNA,SRXN1,MMP-9,and Cleaved Caspase-3 proteins in TE-1 cells.Dual luciferase reporter gene assay was applied to detect the inter-action between LncRNA SOX2-OT and miR-194-5p,and between miR-194-5p and SRXN1.Results The EdU positive cell rate,migration number,invasion number,LncRNA SOX2-OT and SRXN1 mRNAs and proteins,PCNA,SRXN1,and MMP-9 proteins in the sh-SOX2-OT group were lower than those in sh-NC group and control group,apoptosis rate,miR-194-5p,and cleaved Caspase-3 protein expression were higher in sh-NC group and control group(P＜0.05).Compared with sh-SOX2-OT group and sh-SOX2-OT+anti-NC group,the sh-SOX2-OT+anti-miR-194-5p group showed an increase in EdU positive cell rate,migration num-ber,invasion number,SRXN1 mRNA and protein,PCNA,SRXN1,MMP-9 protein expression,and a decrease in apoptosis rate,miR-194-5p,and cleaved Caspase-3 protein expression(P＜0.05).LncRNA SOX2-OT was able to target negative regulation of miR-194-5p,miR-194-5p was able to target negative regulation of SRXN1.Conclusion Knockout of LncRNA SOX2-OT could in-hibit the occurrence and development of ESCC cells,and its mechanism may be achieved by regulating the miR-194-5p/SRXN1 signaling pathway.
郭剑锋;熊玉梅;许芬芬;刘劼
攀钢集团总医院消化科,四川攀枝花 617000攀钢集团总医院消化科,四川攀枝花 617000攀钢集团总医院消化科,四川攀枝花 617000攀钢集团总医院消化科,四川攀枝花 617000
医药卫生
长链非编码RNA性别决定相关基因簇2重叠转录本微小RNA-194-5p硫氧还蛋白-1食管鳞状细胞癌
LncRNAsex determining region Y-box 2 overlapping transcriptmiRNA-194-5pthioredoxin 1esophageal squamous cell carcinoma
《四川医学》 2026 (5)
509-515,7
攀枝花市2022年度市级指导性科技计划项目(编号:2022ZD-S-48)
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