摘要
目的:探讨长链非编码核富含丰富的转录本1(lncRNA NEAT1)在宫颈癌中的表达,及其对宫颈癌细胞HeLa增殖、细胞周期与凋亡的影响。方法:选取2018年3月至2020年3月在四川省人民医院进行外科切除病灶的宫颈癌患者50例作为研究对象,取同期50例因子宫平滑肌瘤(宫颈无任何病变)等良性疾病而切除子宫的正常宫颈组织作为对照,qRT-PCR检测lncRNA NEAT1在肿瘤组织与正常组织中的表达。分析lncRNA NEAT1的表达与宫颈癌患者临床病理参数的关系。细胞实验分为3组:正常组:(不加任何药物,常规培养)、对照组:(细胞转染150 nmol/L的lncRNA NEAT1-siRNA-NC后,常规培养);实验组:(细胞转染150 nmol/L的lncRNA NEAT1-siRNA后,常规培养)。CCK-8法检测各组细胞的增殖;DNA ladder分析各组细胞DNA的损伤情况;流式细胞术检测各组细胞的细胞周期和凋亡率;裸鼠皮下种植瘤模型检测各组细胞的体内生长能力;Western blot检测各组细胞中磷酸化组蛋白(γ-H2AX)、毛细血管扩张性共济失调症突变蛋白激酶(ATM)蛋白表达水平。结果:与正常组织相比,lncRNA NEAT1在肿瘤组织中的表达明显升高。lncRNA NEAT1的相对表达与宫颈癌患者的肿瘤大小、组织学分级相关(均P<0.05);与正常组相比,实验组细胞增殖能力明显下降,DNA出现明显的凋亡“阶梯图谱”,处于G1期细胞的比例明显下降,处于S期细胞的比例明显升高,形成S期阻滞,细胞凋亡率、细胞中γ-H2AX、ATM蛋白的表达明显升高,差异均具有统计意义(均P<0.05)。结论:在宫颈癌组织中lncRNA NEAT1的表达明显升高,沉默其表达后,宫颈癌细胞的HeLa细胞的DNA损伤明显,细胞周期被阻滞于S期,体外增殖与体内生长明显受限,凋亡率明显升高。
Objective:To investigate the expression of long non-coding RNA nuclear-enriched abundant transcript 1(lncRNA NEAT1)in cervical cancer and its effect on proliferation,cell cycle and apoptosis of HeLa cells.Methods:Fifty patients with cervical cancer who underwent surgical resection of the lesions in our hospital from March 2018 to March 2020 were selected as the research objects. Fifty cases of uterine leiomyoma,during the same period,without any cervical lesions and other benign diseases were removed and the uterus were selected as the control objects. qRT-PCR was used to detect the expression of lncRNA NEAT1 in tumor tissues and normal tissues. The relationship between the expression of lncRNA NEAT1 and the clinicopathological parameters of cervical cancer patients was analyzed. The cell experiment was divided into three groups:normal group:(routine culture without any drugs),control group:(cells transfected with 150 nmol/L lncRNA NEAT1-siRNA-NC conventional culture),experimental group:(cells transfected with 150 nmol/L lncRNA NEAT1-siRNA,conventional culture);CCK-8 method was used to detect the proliferation of each group of cells;DNA ladder was used to analyze the DNA damage of each group of cells;flow cytometry was used to detect the cell cycle and apoptosis rate of each group of cells. Nude mice subcutaneous implant tumor model was used to detect the growth in vivo. Western blot was used to detect the protein expression levels phosphorylated histone(γ-H2AX)and ataxia-telangiectasia mutated kinase(ATM)in each group of cells.Results:Compared with normal tissues,the expression of lncRNA NEAT1 in tumor tissues was significantly higher.The relative expression of lncRNA NEAT1 was correlated with the tumor size and histological grade of cervical cancer patients(all P<0.05);compared with the normal group,the cell proliferation ability of the experimental group was significantly decreased,and the DNA showed an obvious "ladder map" of apoptosis,the proportion of cells in G1phase was significantly decreased,the proportion of cells in S phase was significantly increased,the S phase block was formed,the apoptosis rate,the expressions of γ-H2AX and ATM protein in cells were significantly increased,which differences were statistically significant(all P<0.05).Conclusion:The expression of lncRNA NEAT1 is significantly increased in cervical cancer tissues. After silencing its expression,the DNA damage of HeLa cells is obvious,the cell cycle is blocked in S phase,the proliferation in vitro and growth in vivo are significantly restricted,and the apoptosis rate is significantly increased.
作者
李怡琳
梅劼
郭晓霞
雷华江
廖治
凌波
江虹
LI Yilin;MEI Ji;GUO Xiaoxia;LEI Huajiang;LIAO Zhi;LING Bo;JIANG Hong(Sichuan Provincial People's Hospital,Sichuan Academy of Medical Sciences,Chengdu 610000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第17期2107-2112,共6页
Chinese Journal of Immunology
