利用lncRNA和mRNA表达谱揭示热疗对胃癌耐药细胞SGC-7901/DDP增敏的机制  

LncRNA and mRNA Expression Profiling Reveals Mechanism of Hyperthermia Sensitizing Gastric Cancer Resistant Cell SGC-7901/DDP

作　　者：刘佳佳 陈冬梅 蒙洁 王一清 陈彻 王晶 LIU Jia-Jia;CHEN Dong-Mei;MENG Jie;WANG Yi-Qing;CHEN Che;WANG Jing(Molecular Biology Laboratory for Clinical Laboratory Diagnostics,School of Clinical Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;Institute of Reproduction Medicine,First Hospital of Lanzhou University,Lanzhou 730000,China;Key Laboratory of Traditional Chinese Herbs and Prescription Innovation and Transformation of Gansu Province,Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学临床医学院临床检验诊断学分子生物实验室,兰州730000 [2]兰州大学第一医院生殖医学专科研究所,兰州730000 [3]甘肃中医药大学甘肃省中药与方剂创新转化重点实验室,兰州730000

出　　处：《中国生物化学与分子生物学报》2020年第10期1188-1198,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：甘肃省卫生行业科研计划项目(No.GSWSKY2017-27);国家自然科学基金(No.81760835)。

摘　　要：药物耐药导致胃癌(gastric cancer,GC)细胞对化疗的敏感性降低,而热疗(hyperthermia,HT)可以增加化疗的敏感性并引起细胞内基因发生特异性表达。然而,热疗增强细胞SGC-7901/DDP对顺铂(cisplatin,diaminedichloroplatinum,DDP)敏感性的分子机制研究尚缺乏报道。在本研究中,利用MTT实验和流式细胞术分析了对照组、DDP组、HT组和DDP联合HT组细胞SGC-7901/DDP的增殖与凋亡情况。采用高通量微阵列分析和实时定量聚合酶链式反应(qRT-PCR)分析热疗增敏的分子机制。结果显示,与DDP组相比,HT组与DDP联合HT组的细胞增殖被显著抑制(P<0.001),而与DDP联合HT组细胞相比,HT组细胞增殖受抑制更为显著(P<0.05);与DDP组相比,HT组、DDP联合HT组细胞发生早期凋亡的比例显著增加(P<0.001),且DDP联合HT组明显高于HT组(P<0.05),表明HT增强了顺铂对细胞SGC-7901/DDP的敏感性。相比对照组,在DDP联合HT干预后,LINC00161和TCONS_00018082上调(P<0.01),LINC00473和TCONS_00015171下调(P<0.01);另外,DDP联合HT组比对照组中的差异表达mRNA显著富集在凋亡信号通路、TGF-β信号通路和Notch信号通路(P<0.05)。本研究提示,热疗可能通过调控上述lncRNA和相关通路增强细胞SGC-7901/DDP对顺铂的敏感性。Drug resistance leads to reduced sensitivity of gastric cancer(GC)cell to chemotherapy,while hyperthermia(HT)can increase sensitivity and cause specific expression of genes.However,the molecular mechanism of HT sensitization to diaminedichloroplatinum(DDP)in SGC-7901/DDP has not been studied.In this study,MTT assay and flow cytometry were used to analyze proliferation and apoptosis of SGC-7901/DDP in control group,DDP group,HT group,DDP combined with HT group.Microarray analysis and quantitative real-time polymerase chain reaction(qRT-PCR)were performed to explore mechanism of HT sensitization.The results showed that cell proliferation of HT group and DDP combined HT group was significantly inhibited compared with cisplatin group(P<0.001),and the cell proliferation of the DDP combined HT group was more significantly inhibited than the HT group(P<0.05);Compared with DDP group,the early apoptosis rate of HT group and DDP combined HT group was significantly increased(P<0.001),and DDP combined with HT group was significantly higher than HT group(P<0.05),indicating that HT enhanced the sensitivity of cisplatin to cell SGC-7901/DDP.Compared with the control group,LINC00161 and TCONS_00018082 were up-regulated(P<0.01),LINC00473 and TCONS_00015171 were down-regulated(P<0.01)after DDP combined with HT intervention SGC-7901/DDP.In addition,the differentially expressed mRNA after DDP combined with HT group was significantly enriched in apoptosis signaling pathway,TGF-βsignaling pathway and Notch signaling pathway(P<0.05).In summary,HT can enhance sensitivity of SGC-7901/DDP to DDP by regulating related apoptotic genes and pathways.

关 键 词：胃癌 热疗 顺铂 敏化作用 长链非编码RNA 

分 类 号：Q354[生物学—遗传学]