摘要
目的 探讨长链非编码RNA OIP5反义RNA 1(OIP5-AS1)靶向调控微小RNA-200b-3p(miR-200b-3p)在食管鳞癌(ESCC)细胞增殖和凋亡中的作用。方法 采用实时荧光定量PCR(qPCR)检测ESCC组织和细胞的OIP5-AS1和miR-200b-3p水平。双荧光素酶报告基因实验鉴定OIP5-AS1与miR-200b-3p之间的相互作用。将靶向OIP5-AS1的小干扰RNA序列si-OIP5-AS1、miR-200b-3p抑制物(inhibitor)单独或共转染EC109细胞,qPCR验证转染效果后采用MTT法和Annexin-V/PI双染流式细胞术检测EC109细胞的增殖和凋亡情况。结果 与癌旁组织比较,ESCC组织的OIP5-AS1水平升高而miR-200b-3p水平降低(P<0.05),且两者水平呈负相关性(r=-0.415,P<0.001)。与HET-1A细胞相比,ESCC细胞的OIP5-AS1水平上调而miR-200b-3p水平下调,差异均有统计学意义(P<0.05)。生物信息学分析和双荧光素酶报告基因实验结果显示OIP5-AS1可靶向结合miR-200b-3p。与未转染细胞相比,EC109细胞转染si-OIP5-AS1后的凋亡率升高且细胞活力减弱,而转染miR-200b-3p inhibitor后的凋亡率降低且细胞活力增强,上述差异有统计学意义(P<0.05);共转染后的凋亡率和细胞活力与未转染细胞的差异无统计学意义(P>0.05)。结论 OIP5-AS1通过抑制miR-200b-3p进而诱导ESCC细胞增殖和抑制细胞凋亡,并在ESCC的发展中起到肿瘤启动因子的作用,OIP5-AS1/miR-200b-3p轴有望成为ESCC的防治靶点。
Objective To explore the exact role of long non-coding RNA OPA-interacting protein 5 antisense transcript 1(OIP5-AS1) targeting microRNA-200b-3p(miR-200b-3p) in the proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC) cells. Methods Levels of OIP5-AS1 and miR-200b-3p in ESCC tissues and cell lines were detected by quantitative real-time PCR(qPCR). Dual luciferase reporter assay was performed to analyze the interaction between OIP5-AS1 and miR-200b-3p. EC109 cells were exposed to small interfering RNA sequence si-OIP5-AS1 targeting OIP5-AS1 alone, miR-200b-3p inhibitor alone or their combined transfection. Transfection effect was verified by qPCR. The proliferation and apoptosis of EC109 cells were detected by MTT method and flow cytometry with Annexin V FITC/PI double staining. Results Compared with the adjacent tissues, the OIP5-AS1 level increased and the miR-200b-3p level decreased in ESCC tissues(P<0.05), and there was a negative correlation between the two levels(r=-0.415, P<0.001). Compared with HET-1A cells, the OIP5-AS1 level was up-regulated, and the miR-200b-3p level was down-regulated in ESCC cells(P<0.05). Bioinformatics analysis and double luciferase reporter gene experiment showed that OIP5-AS1 could target miR-200b-3p. Compared with untransfected cells, EC109 cells transfected with si-OIP5-AS1 had higher apoptotic rate and lower cell viability, while cells transfected with miR-200b-3p inhibitor had lower apoptotic rate and higher cell viability, the difference was statistically significant(P<0.05). No significant differences in apoptotic rate and cell viability were observed between co-transfected cells and non-transfected cells(P>0.05). Conclusion OIP5-AS1 can induce ESCC cell proliferation and inhibit cell apoptosis by inhibiting miR-200b-3p, and play the role of tumor initiating factor in the development of ESCC. OIP5-AS1/miR-200b-3p axis is expected to become the target of ESCC prevention and treatment.
作者
谢宙航
杨朝坤
解少强
XIE Zhouhang;YANG Chaokun;XIE Shaoqiang(Department of Cardiothoracic Surgey,The Second People's Hospital of Yibin,Yibin 644000,China)
出处
《临床肿瘤学杂志》
CAS
2022年第12期1065-1071,共7页
Chinese Clinical Oncology
关键词
食管鳞癌
OIP5反义RNA
1
微小RNA-200b-3p
增殖
凋亡
Esophageal squamous cell carcinoma
OPA-interacting protein 5 antisense transcript 1
MicroRNA-200b-3p
Proliferation
Apoptosis
