摘要
目的:探究长链非编码RNA(long non-coding RNA,LncRNA)肺癌相关转录本1(lung cancer-associated transcript 1,LUCAT1)在宫颈癌细胞增殖、转移及顺铂(DDP)耐药中的作用及可能机制。方法:收集30例宫颈癌患者肿瘤组织和癌旁组织,采用实时荧光定量PCR(qPCR)检测LncLUCAT1的表达;宫颈癌SiHa细胞作为实验细胞系,siRNA干扰SiHa细胞中LncLUCAT1表达后,通过MTT与集落克隆实验检测细胞的增殖能力,Transwell实验检测细胞的侵袭迁移能力;在SiHa/DDP细胞中干扰和过表达LncLUCAT1后,使用qPCR验证转染效果,MTT检测细胞IC50的变化,通过Western blot检测SiHa/DDP细胞中AKT与p-AKT蛋白的表达。结果:相比宫颈癌旁组织,宫颈癌组织中LncLUCAT1呈现高表达,MTT、集落克隆及Transwell结果显示下调LncLUCAT1抑制SiHa细胞增殖及侵袭迁移能力。qPCR结果显示在SiHa/DDP细胞中LncLUCAT1表达显著高于SiHa细胞(P<0.05)。干扰LncLUCAT1表达可增加SiHa/DDP对DDP的敏感度,过表达LncLUCAT1可降低SiHa/DDP对DDP的敏感度。Western blot结果显示干扰LncLUCAT1表达抑制p-AKT蛋白的表达,过表达LncLUCAT1促进p-AKT蛋白的表达。结论:LncLUCAT1可促进宫颈癌SiHa细胞的增殖、转移及顺铂耐药,其机制可能与LncLUCAT1调控AKT蛋白磷酸化的表达相关。
Objective:To investigate the effect and possible mechanism of lung cancer associated transcript 1(LUCAT1)of long non-coding RNA on proliferation,metastasis and cisplatin resistance(DDP)in cervical cancer cells.Methods:The expression of LncLUCAT1 in cancer and pericarcinomatous tissue of 30 patients with cervical cancer was detected by real-time quantitative PCR(qPCR).Cervical cancer SiHa cells were used as experimental cell lines.After siRNA interfering with expression of LncLUCAT1 in SiHa cells,the proliferation ability of cells was detected by MTT and clonogenic assay,and the invasion and migration ability of cells were measured by transwell assay.After LncLUCAT1 being interfered and overexpressed in SiHa/DDP cells,the transfection effect was verified by qPCR,the changes of IC50 were detected by MTT,and the expressions of AKT and p-Akt in SiHa/DDP cells were detected by Western blot.Results:Compared with para-cervical tissues,LncLUCAT1 was highly expressed in cervical cancer tissues.MTT,clonogenic assay and transwell migration assay showed that the proliferation,invasion and migration ability of SiHa cells inhibited by down-regulated LncLUCAT1.qPCR results showed that LncLUCAT1 expression in SiHa/DDP cells was significantly higher than that in SiHa cells(P<0.05).Interference of LncLUCAT1 expression increased the sensitivity of SiHa/DDP to DDP,while overexpression of LncLUCAT1 reduced the sensitivity of SiHa/DDP to DDP.Western blot results showed that interference of LncLUCAT1 expression inhibited p-AKT protein expression,while overexpression of LncLUCAT1 promoted p-AKT protein expression.Conclusion:LncLUCAT1 could regulate the phosphorylation of AKT and reduce the sensitivity of cervical cancer SiHa cells to cisplatin.
作者
魏丽
陈玉忠
杜军
李艳
张慧慧
李多杰
WEI Li;CHEN Yuzhong;DU Jun;LI Yan;ZHANG Huihui;LI Duojie(Department of Gynecological Oncology,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Surgical Oncology,the First Affiliated Hospital of Bengbu Medical College;Department of Radiotherapy,the First Affiliated Hospital of Bengbu Medical College)
出处
《包头医学院学报》
CAS
2023年第5期22-28,共7页
Journal of Baotou Medical College
基金
蚌埠医学院自然科学项目重点项目(2020byzd107),蚌埠医学院科研计划项目(2020byzd133)。
