Lnc RNA HNF1A-AS1靶向miR-32-5p调控肺癌对顺铂的耐药性  

Effects of Lnc RNA HNF1A-AS1 on lung cancer resistance to cisplatin by targeting miR-32-5p

作　　者：张艳丽[1] 唐文慧[3] 苏军[2] 马红霞[2] ZHANG Yanli;TANG Wenhui;SU Jun(Department of Respiratory Diseases,TCM Hospital Affiliated to Xinjiang Medical University,Xinjiang,Urumqi 830000,China;不详)

机构地区：[1]新疆医科大学附属中医医院呼吸科,乌鲁木齐市830000 [2]新疆医科大学附属中医医院呼吸与危重症医学科,乌鲁木齐市830000 [3]北京市大兴区人民医院呼吸与危重症医学科

出　　处：《河北医药》2021年第21期3229-3232,共4页Hebei Medical Journal

摘　　要：目的探讨长链非编码RNA HNF1A-AS1对顺铂耐药肺癌细胞凋亡、耐药性的影响机制。方法运用MTT法检测顺铂耐药肺癌细胞H520/DDP的IC50。将si-NC组(转染si-NC)、si-HNF1A-AS1组(转染si-HNF1A-AS1)、miR-NC组(转染miR-NC)、miR-32-5p组(转染miR-32-5p mimics)、si-HNF1A-AS1+anti-miR-NC组(共转染si-HNF1A-AS1和anti-miR-NC)和si-HNF1A-AS1+anti-miR-32-5p组(共转染si-HNF1A-AS1和anti-miR-32-5p)均用脂质体法转染至H520/DDP细胞。RT-qPCR实验检测细胞中HNF1A-AS1、miR-32-5p的mRNA的表达;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测细胞的荧光活性。结果与肺鳞状细胞癌细胞H520相比,H520/DDP细胞的IC50显著升高,细胞凋亡率显著降低(P<0.05),成功构建顺铂耐药H520细胞;H520/DDP细胞中HNF1A-AS1的表达明显升高,miR-32-5p的表达明显降低(P<0.05);抑制HNF1A-AS1、过表达miR-32-5p均可明显下调H520/DDP细胞的IC50,促进凋亡;miR-32-5p可抑制野生型HNF1A-AS1细胞的荧光活性,并负向调控其表达。抑制miR-32-5p可逆转抑制HNF1A-AS1对H520/DDP细胞的增敏和促凋亡作用。结论长链非编码RNA HNF1A-AS1可通过靶向负调控miR-32-5p的表达调控顺铂耐药肺癌细胞的顺铂敏感性。Objective To investigate the effects of Lnc RNA HNF1A-AS1 on apoptosis and drug tolerance of lung cancer cells to cisplatin by targeting miR-32-5p.Methods The IC50 of H520/DDP in cisplatin-resistant lung cancer cells was detected by MTT assay.Those in siNC group(transfected siNC),siHNF1A-AS1 group(transfected siHNF1A-AS1),miRNC group(transfected miR-NC),miR-32-5p group(transfected miR-32-5p mimics),siHNF1A-AS1+anti-miR-NC group(co-transfected siHNF1A-AS1 and anti-miR-NC),siHNF1A-AS1+anti-miR-32-5p groups(co-transfected siHNF1A-AS1 and anti-miR-32-5p)were transfected into H520/DDP cells by liposome method.RTqPCR assay was used to detect the expression of HNF1A-AS1 and miR-32-5p mRNA.Flow cytometry was used to detect apoptosis and double luciferase reporter assay was used to detect the fluorescence activity of the cells.Results Compared with that in lung squamous cell carcinoma cell line H520,the IC50 of H520/DDP cells was significantly increased,however,the apoptosis rate was significantly decreased(P<0.05).Cisplatin resistant H520 cells were successfully constructed.The expression levels of HNF1A-AS1 in H520/DDP cells wee significantly increased,but the expression levels of miR-32-5p were significantly decreased(P<0.05).The inhibition of HNF1A-AS1 and overexpression of miR-32-5p cloud significantly down-regulate the IC50 of H520/DDP cells and promote cell apoptosis.In addition the miR-32-5p inhibited the fluorescent activity of wild type HNF1A-AS1 cells and negatively regulated its expression.Moreover the inhibition of miR-32-5p reversibly inhibited the sensitization and pro-apoptotic effects of HNF1A-AS1 on H520/DDP cells.Conclusion The long chain non-coding RNA HNF1A-AS1 can regulate the cisplatin sensitivity of cisplatin resistant lung cancer cells by targeting negative regulation of the expression of miR-32-5p.

关 键 词：长链非编码RNA HNF1A-AS1 miR-32-5p 肺癌 顺铂敏感性 

分 类 号：R734.2[医药卫生—肿瘤]