ITGB2反义RNA 1对非小细胞肺癌细胞顺铂敏感性的影响  

EffectsI of ITGB2 antisense RNA 1 on the sensitivity of non-small cell lung cancer cells to cisplatin

作　　者：陈慧勇[1] 王开雷 万仁平[1] 李友涛[1] 王林辉[1] CHEN Huiyong;WANG Kailei;WAN Renping;LI Youtao;WANG Linhui(Department of Thoracic Surgery,Yuebei People’s Hospital,Shaoguan 512026,China)

机构地区：[1]粤北人民医院胸外科,广东韶关512026

出　　处：《临床肿瘤学杂志》2024年第12期1152-1158,共7页Chinese Clinical Oncology

基　　金：韶关市科技局课题(230330178036323)。

摘　　要：目的 探讨长链非编码RNA ITGB2反义RNA 1(ITGB2-AS1)对非小细胞肺癌细胞顺铂敏感性的影响。方法 收集非小细胞肺癌患者及顺铂耐药患者的癌旁组织和癌组织,荧光原位杂交技术检测组织中ITGB2-AS1的表达水平。实时荧光定量PCR(qPCR)检测顺铂耐药细胞A549/DDP和H1975/DDP中ITGB2-AS1的表达。构建靶向ITGB2-AS1的小发夹RNA序列(sh-ITGB2-AS1)和阴性对照序列(sh-NC)并转染A549/DDP和H1975/DDP细胞,分为Control组、Cisplatin组、Cisplatin+sh-NC组和Cisplatin+sh-ITGB2-AS1组,qPCR检测sh-ITGB2-AS1的干扰效率,采用CCK-8和TUNEL实验检测细胞活力和凋亡水平,Transwell实验检测细胞的迁移和侵袭能力,Western blot检测细胞中多药耐药蛋白1(MRP1)和P-糖蛋白(P-gp)的表达水平。结果 与癌旁组织相比,癌组织中ITGB2-AS1表达水平升高,且化疗耐药患者癌组织中ITGB2-AS1表达水平最高。与A549和H1975细胞相比,A549/DDP和H1975/DDP中ITGB2-AS1表达水平升高。转染sh-ITGB2-AS1抑制了A549/DDP和H1975/DDP细胞中ITGB2-AS1表达水平。与Control组相比,Cisplatin组和Cisplatin+sh-NC组的细胞活力、凋亡水平和迁移侵袭能力无显著变化,而与Cisplatin+sh-NC组相比,Cisplatin+sh-ITGB2-AS1组的细胞活力显著下降,细胞凋亡水平升高,迁移侵袭能力下降。与Control组、Cisplatin组和Cisplatin+sh-NC组相比,Cisplatin+sh-ITGB2-AS1组细胞的MRP1和P-gp蛋白表达水平下降。结论 肺癌中ITGB2-AS1表达上调,且抑制ITGB2-AS1表达能够增强顺铂耐药细胞对于顺铂药物的敏感性。Objective To investigate the role of ITGB2 antisense RNA 1(ITGB2-AS1)on cisplatin drug sensitivity in non-small cell lung cancer cells.Methods Paracancerous and cancerous tissues from non-small cell lung cancer patients and cisplatin-resistant patients were collected,and the expression level of ITGB2-AS1 in the tissues was detected by fluorescence in situ hybridization.Cisplatin-resistant cell lines A549/DDP and H1975/DDP were cultured,and ITGB2-AS1 expression was detected by real-time fluorescence quantitative PCR(qPCR)in A549/DDP and H1975/DDP cells.Small hairpin RNA sequence targeting ITGB2-AS1(sh-ITGB2-AS1)and a negative control sequence(sh-NC)were constructed and transfected into A549/DDP and H1975/DDP cells.Cells were divided into Control group,Cisplatin group,Cisplatin+sh-NC group,and Cisplatin+sh-ITGB2-AS1 group.Moreover,qPCR was performed to detect interference efficiency of sh-ITGB2-AS1.Cell viability and apoptosis levels were detected by CCK-8 and TUNEL assays.Cell abilities of migration and invasion were detected by Transwell assay,and multidrug resistance protein 1(MRP1)and P-glycoprotein(P-gp)expression levels were detected in cells by Western blot.Results ITGB2-AS1 expression levels were elevated in cancer tissues compared with paracancerous tissues,and were highest in cancer tissues of chemoresistant patients.Compared with A549 and H1975 cells,ITGB2-AS1 expression levels were elevated in A549/DDP and H1975/DDP cells.Transfection of sh-ITGB2-AS1 inhibited ITGB2-AS1 expression levels in A549/DDP and H1975/DDP cells.There were no significant changes in cell viability,apoptosis level and abilities of migration and invasion in the Cisplatin and Cisplatin+sh-NC groups compared to the Control group.The decreased cell viability,elevated apoptosis level and weakened abilities of migration and invasion were found in Cisplatin+sh-ITGB2-AS1 group as compared with Cisplatin+sh-NC group.Expression levels of MRP1 and P-gp proteins were decreased in the cells of Cisplatin+sh-ITGB2-AS1 group compared with Control,

关 键 词：非小细胞肺癌 ITGB2反义RNA 1 顺铂 耐药 

分 类 号：R374.2[医药卫生—病原生物学]