摘要
目的观察长链非编码RNA(lncRNA)靶向微小RNA(miR)-130a-3p基因对骨关节炎(OA)软骨细胞增殖及炎性因子分泌的影响。方法将中南大学湘雅医学院附属海口医院从2018年1月至2020年1月收治的OA患者30例纳入研究。分别采集OA以及正常软骨细胞,通过实时荧光聚合酶链反应(RT-PCR)检测不同软骨细胞中的lncRNA同源基因转录反义RNA(HOTAIR)以及miR-130a-3p表达水平。此外,于OA软骨细胞中转染siRNA-lncRNA HOTAIR或LV-lncRNA HOTAIR,采用细胞增殖实验(CCK-8)法完成细胞增殖情况的检测,并以RT-PCR法检测炎性因子分泌情况。计数资料以%表示,采用χ2检验。符合正态分布的计量资料以均数±标准差(±s)表示,采用t检验。结果OA软骨细胞中的lncRNA HOTAIR mRNA相对表达量为3.01±0.42,明显高于正常软骨细胞(1.08±0.23),而miR-130a-3p mRNA相对表达量为0.48±0.11,明显低于正常软骨细胞(1.00±0.01),差异均有统计学意义(P值均<0.05)。转染si-lncRNA HOTAIR组OA软骨细胞的增殖能力相较于转染si-NC组明显增加,而转染LV-lncRNA HOTAIR组OA软骨细胞的增殖能力相较于转染LV-NC组显著下降,差异均有统计学意义(P值均<0.05)。转染si-lncRNA HOTAIR组OA软骨细胞的miR-130a-3p表达水平显著升高,而转染LV-lncRNA HOTAIR组OA软骨细胞的miR-130a-3p表达水平显著下降,且经Pearson相关分析显示,在OA软骨细胞中,lncRNA HOTAIR和miR-130a-3p表达水平呈明显负相关关系,差异有统计学意义(r=-0.912,P<0.05)。转染si-lncRNA HOTAIR后的OA软骨细胞肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)mRNA相对表达量分别为0.47±0.10、0.52±0.13,均明显低于转染LV-lncRNA HOTAIR后(0.88±0.19、0.93±0.15),差异有统计学意义(P值均<0.05)。结论lncRNA HOTAIR通过靶向调控miR-130a-3p基因,继而影响OA软骨细胞的增殖,并具有一定的炎性反应调控作用。
Objective To investigate and analyze the effects of microRNA(miR)-130a-3p targeted by long non-coding RNA(lncRNA)on proliferation and inflammatory factor secretion of osteoarthritis(OA)chondrocytes.Methods A total of 30 patients with OA admitted to the Haikou Hospital from February 2018 to February 2020 were included in the study.OA and normal chondrocytes were collected respectively,and the expression levels of lncRNA HOX transcript antisense RNA(HOTAIR)and miR-130a-3p in different chondrocytes were detected by real-time fluorescence polymerase chain reaction(RT-PCR).In addition,siRNA-lncRNA HOTAIR or LV-lncRNA HOTAIR was transfected into OA chondrocytes,and cell proliferation was examined by cell counting kit-8(CCK-8)method.The inflammatory factor secretion was determined by RT-PCR method.The data were analyzed by the SPSS 22.0 software,count data were expressed as%,and theχ2 test was done.The measurement data were expressed as mean±standard deviation,and t test was carried out.P<0.05 indicated a statistically significant difference.Results The relative expression level of lncRNA HOTAIR mRNA in OA chondrocytes was 3.01±0.42,significantly higher than that in normal chondrocytes(1.08±0.23),and the relative expression level of miR-130a-3p mRNA was 0.48±0.11,significantly lower than that in normal chondrocytes(1.00±0.01)(all P<0.05).The proliferation ability of OA chondrocytes in the si-lncRNA HOTAIR transfection group was significantly increased as compared with that in the Si-NC transfection group,and that of OA chondrocytes in the lentivirus-long non-coding RNA(LV-lncRNA)HOTAIR transfection group was significantly decreased as compared with that in the LV-NC transfection group(all P<0.05).The expression level of miR-130a-3p in OA chondrocytes transfected with silencing-long non-coding RNA(si-lncRNA)HOTAIR was significantly increased,and that in OA chondrocytes transfected with LV-lncRNA HOTAIR was significantly decreased.The Pearson correlation analysis showed that lncRNA HOTAIR and miR-130a-3p expression
作者
王和杰
范忠诚
金旭红
卓泽铭
邢势
Wang Hejie;Fan Zhongcheng;Jin Xuhong;Zhuo Zeming;Xing Shi(Orthopaedic Medicine Center of Haikou Hospital Affiliated to Xiangya Medical College of Zhongnan University,Haikou 572000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第5期871-873,共3页
Chinese Journal of Experimental Surgery
