摘要
目的探讨转录失调的长链非编码RNA(LncRNA)对耐药非小细胞肺癌(NSCLC)细胞生物学特性的影响及作用机制。方法选择2019年4月至2021年2月秦皇岛市第一医院收治的对化学治疗药物耐药的NSCLC患者61例为研究对象。采用经皮肺穿刺的方法获取患者的肺癌组织和癌旁组织,并采用转录组测序及LncRNA表达定量分析方法筛选出2种组织中差异表达的LncRNA。将肺癌组织和癌旁组织中LncRNA表达量比较差异有统计学意义的HOXA11-AS转染A549/DDP细胞(HOXA11-AS转染组),另取A549/DDP细胞转染空载体作为对照组。采用噻唑蓝法检测2组细胞的增殖能力,划痕愈合实验检测2组细胞的迁移能力,Transwell法检测2组细胞的侵袭能力,Western blot法测定2组细胞中磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(AKT)蛋白的表达。结果共筛选出311个差异表达LncRNA,共表达最多的前10个LncRNA中,HOXA11-AS在肺癌组织中的表达量显著高于癌旁组织(P<0.05)。培养1 d时,2组细胞的增殖能力比较差异无统计学意义(P>0.05);培养2、3、4、5 d时,HOXA11-AS转染组细胞的增殖能力显著高于对照组(P<0.05)。培养0 h时,2组细胞的迁移率比较差异无统计学意义(P>0.05);培养24 h时,HOXA11-AS转染组细胞的迁移率显著高于对照组(P<0.05)。HOXA11-AS转染组穿过基底膜的细胞数显著多于对照组(P<0.05)。HOXA11-AS转染组细胞中PI3K、AKT蛋白相对表达量显著高于对照组(P<0.05)。结论转录失调的LncRNA中的HOXA11-AS参与了NSCLC患者对顺铂耐药的发生与发展。HOXA11-AS可促进耐药细胞株A549/DDP细胞的增殖、迁移和侵袭,机制可能与PI3K/AKT信号通路激活有关。
Objective To explore the effect of dystranscriptional long non-coding RNA(LncRNA)on biological characteristics of drug resistant non-small cell lung cancer(NSCLC)cells and its mechanism.Methods Sixty-one NSCLC patients who were resistant to chemotherapy drugs admitted to the First Hospital of Qinhuangdao from April 2019 to February 2021 were selected as the research subjects.The lung cancer tissue and adjacent tissue were obtained by percutaneous lung puncture,and the differentially expressed LncRNAs in the above two tissues were screened by transcriptome sequencing and quantitative analysis of LncRNA expression.The differentially expressed LncRNA HOXA11-AS with significant differences in expression levels between lung cancer tissue and adjacent tissues were transfected into A549/DDP cells(HOXA11-AS transfection group),another A549/DDP cells were taken and transfected with empty vector as the control group.The proliferation ability of cells in the two groups was determined by methyl thiazolyl tetrazolium bromide method,the migration ability of cells in the two groups was determined by scratch healing experiment,the invasion ability of cells in the two groups was determined by Transwell method,and the expressions of phosphatidylinositol-3-kinase(PI3K)and protein kinase B(AKT)protein in cells in the two groups were measured by Western blot method.Results A total of 311 differentially expressed LncRNAs were selected,and among the top 10 LncRNAs with the highest co-expression,the expression level of HOXA11-AS in lung cancer tissue was significantly higher than that in adjacent tissues(P<0.05).At 1 d of cultivation,there was no significant difference in the proliferation ability of cells between the two groups(P>0.05);at 2,3,4,5 d of cultivation,the proliferation ability of cells in the HOXA11-AS transfection group was significantly higher than that in the control group(P<0.05).At 0 h of cultivation,there was no significant difference in the cell migration rate between the two groups(P>0.05);at 24 h of cultivation,the cell migration rate in the HOXA11-AS transfection group was significantly higher than that in the control group(P<0.05).The number of cells passing through the basement membrane in the HOXA11-AS transfection group was significantly more than that in the control group(P<0.05).The relative expressions of PI3K and AKT protein in cells in the HOXA11-AS transfection group were significantly higher than those in the control group(P<0.05).Conclusion HOXA11-AS,one of the LncRNA with transcriptional disorder,participates in the occurrence and development of cisplatin resistance in NSCLC patients.It can promotes the proliferation,migration and invasion of drug-resistant cell line A549/DDP,and the mechanism may be related to the activation of the PI3K/AKT signaling pathway.
作者
贾瑞
王超
李健
JIA Rui;WANG Chao;LI Jian(Department of Thoracic Surgery,First Hospital of Qinhuangdao,Qinhuangdao 066000,Hebei Province,China)
出处
《新乡医学院学报》
CAS
2023年第5期416-420,共5页
Journal of Xinxiang Medical University
基金
秦皇岛市科学技术研究与发展计划项目(编号:202004A103)。
