麦门冬汤合千金苇茎汤对小鼠Lewis原位肺癌lncRNA,mRNA表达谱的影响
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摘要
目的:通过对Lewis肺癌原位移植小鼠肿瘤组织长链非编码RNA(long noncoding RNA,lncRNA)和mRNA表达谱的检测,探讨麦门冬汤合千金苇茎汤(金方)抗肺癌的作用机制。方法:C57BL/6小鼠随机分为正常组、模型组、金方组(20 g·kg-1·d-1),构建小鼠Lewis原位肺癌模型成功后,次日用金方灌胃开始治疗。应用基因芯片技术,检测与金方抗肺癌作用密切相关的差异lncRNA,mRNA,进行聚类分析,并利用t检验和差异表达的倍数变化,筛选出关键lncRNA,mRNA。运用生物信息学方法预测差异lncRNA调控的靶基因,并对其进行功能和通路分析。应用病理组织学技术,检测各组小鼠肿瘤组织光镜下的差异表现。结果:lncRNA,mRNA芯片杂交结果显示,金方调控了887个lncRNA差异表达,其中上调442个,下调445个(P<0.05)。差异表达的mRNA共610个,其中上调376个,下调234个(P<0.05)。使用GO分析方法,发现下调或上调的靶基因主要参与了细胞代谢调控、信号通路调控等生物学过程(biological process,BP)。靶基因分子功能(molecular function,MF)分析显示,下调或上调的靶基因主要具有DNA结合、蛋白质结合、受体结合和转录因子活性等功能。信号通路预测显示金方可调控Janus激酶/信号转导与转录激活因子(JAK/STAT)等与肺癌的发生发展密切相关的信号通路。病理组织学检查结果证实,与模型组比较,金方能明显改善小鼠肺组织的病理改变。结论:金方可能通过调控多种lncRNA和mRNA的表达及下游相关信号通路来发挥其抗肺癌的作用。
Objective:To explore the anti-lung cancer mechanism of Maimendong Tang and Qianjin Weijingtang(Jin Fang) by detecting the expression profiles of long noncoding RNA(lncRNA) and mRNA in mice tumor tissues of orthotopic Lewis lung cancer model.Method:C57 BL/6 mice were randomly divided into normal group,model group and Jin Fang group(20 g·kg-1·d-1).After successful establishment of Lewis lung cancer model in situ in mice,Jin Fang was given orally the next day after treatment.Using gene chip technology,differential lncRNA and mRNA closely related with Jin Fang' s anti-lung cancer effect were detected,and cluster analysis was performed.The key lncRNA and mRNA were screened out by t-test and fold change of differential expression.Bioinformatic methods were used to predict target genes regulated by differential lncRNA,and functional and pathway analysis was performed.The histopathological technique was used to detect the differences in the tumor tissue of each group under light microscope.Result:lncRNA and mRNA chip hybridization results showed that Jin Fang regulated differential expressions of 887 lncRNA,in which 442 were up-regulated and 445 were down-regulated(P<0.05).There were 610 differential mRNA expressions,in which 376 were up-regulated and 234 were down-regulated(P<0.05).GO analysis showed that down-regulated or up-regulated target genes were mainly involved in biological processes(BP),such as cell metabolism regulation and signal pathway regulation.Molecular function(MF) analysis showed such functions DNA binding,protein binding,receptor binding and transcription factor activity in down-regulated or up-regulated target genes.Target genes were involved in multiple biological processes,cellular components and molecular functions.Signal pathway predictions indicated that Jin Fang can regulate the Janus kinase/signal transducer and activator of tran-ions(JAK/STAT) and other signaling pathways closely related to the development of lung cancer.The results of histopathological examination confirmed that Jin Fang could significantly improve the pathological changes of lung tissues in the mice compared with the model group.Conclusion:Jin Fang may exert its anti-lung cancer effect by regulating the expressions of multiple lncRNAs and mRNAs,and down-regulating related signaling pathways.
Objective:To explore the anti-lung cancer mechanism of Maimendong Tang and Qianjin Weijingtang(Jin Fang) by detecting the expression profiles of long noncoding RNA(lncRNA) and mRNA in mice tumor tissues of orthotopic Lewis lung cancer model.Method:C57 BL/6 mice were randomly divided into normal group,model group and Jin Fang group(20 g·kg-1·d-1).After successful establishment of Lewis lung cancer model in situ in mice,Jin Fang was given orally the next day after treatment.Using gene chip technology,differential lncRNA and mRNA closely related with Jin Fang' s anti-lung cancer effect were detected,and cluster analysis was performed.The key lncRNA and mRNA were screened out by t-test and fold change of differential expression.Bioinformatic methods were used to predict target genes regulated by differential lncRNA,and functional and pathway analysis was performed.The histopathological technique was used to detect the differences in the tumor tissue of each group under light microscope.Result:lncRNA and mRNA chip hybridization results showed that Jin Fang regulated differential expressions of 887 lncRNA,in which 442 were up-regulated and 445 were down-regulated(P<0.05).There were 610 differential mRNA expressions,in which 376 were up-regulated and 234 were down-regulated(P<0.05).GO analysis showed that down-regulated or up-regulated target genes were mainly involved in biological processes(BP),such as cell metabolism regulation and signal pathway regulation.Molecular function(MF) analysis showed such functions DNA binding,protein binding,receptor binding and transcription factor activity in down-regulated or up-regulated target genes.Target genes were involved in multiple biological processes,cellular components and molecular functions.Signal pathway predictions indicated that Jin Fang can regulate the Janus kinase/signal transducer and activator of tran-ions(JAK/STAT) and other signaling pathways closely related to the development of lung cancer.The results of histopathological examination confirmed that Jin Fang could significantly improve the pathological changes of lung tissues in the mice compared with the model group.Conclusion:Jin Fang may exert its anti-lung cancer effect by regulating the expressions of multiple lncRNAs and mRNAs,and down-regulating related signaling pathways.
引文
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[12]姜雪莲,姚逸临,罗斌,等.低剂量化疗联合中医药治疗肺癌[J].长春中医药大学学报,2018,34(2):381-383.
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[20]YANG Y,LI H,HOU S,et al.The noncoding RNAexpression profile and the effect of lncRNA AK126698on cisplatin resistance in non-small-cell lung cancer cel[J].PLo S One,2013,8(5):1-12.
[21]TANG Q,NI Z,CHENG Z,et al.Three circulating long non-coding RNAs act as biomarkers for predicting NSCLC[J].Cell Physiol Biochem,2015,37(3):1002-1009.
[22]郑璐玉,熊飞,詹7),等.麦门冬汤合千金苇茎汤提取部位对非小细胞肺癌H460细胞毒作用的研究[J].中国实验方剂学杂志,2010,16(3):60-63.
[23]施文,李俊生.ADAM家族与肿瘤关系的研究现状[J].实用癌症杂志,2008,23(6):671-673.
[24]ZHOU B B,Fridman J S,LIU X,et al.ADAMproteases,Erb B pathways and cancer[J].Expert Opin Investig Drugs,2005,14(6):591-606.
[25]Groner B,Von M V.Jak Stat signaling and cancer:Opportunities,benefits and side effects of targeted inhibition[J].Mol Cell Endocrinol,2017,451(1):1-14.
[26]Harada D,Takigawa N,Kiura K.The role of STAT3 in non-small cell lung cancer[J].Cancers,2014,6(2):708-722.
[27]张宏方,吴芳,环诚,等.调衡方对Lewis肺癌细胞增殖及JAK-STAT信号通路的影响[J].辽宁中医药大学学报,2013,15(3):16-19.
[2]韩荣龙,周家明,胡陵静.癌康宁合剂对晚期非小细胞肺癌化疗后患者维持治疗的临床评价[J].中国实验方剂学杂志,2018,24(19):201-206.
[3]李白坤,湛宇灿,李庆林,等.中医药治疗肺癌作用机制研究进展[J].安徽中医药大学学报,2018,37(3):90-93.
[4]朱赛君,许尤琪.中药复方治疗肺癌作用机理的实验研究进展[J].中医药导报,2018,24(12):57-61.
[5]王珊珊,郭茗,朱垚,等.国医大师周仲瑛教授辨治肺癌经验[J].中华中医药杂志,2015,30(12):4332-4335.
[6]高浩学.麦门冬合千金苇茎汤抗肺癌效应物质基础及其苇茎的化学成分研究[D].南京:南京中医药大学,2009.
[7]李炜,唐于平,高浩学,等.麦门冬合千金苇茎汤效应部位的化学成分[J].中国实验方剂学杂志,2010,16(18):78-81.
[8]熊飞,周宗剑,姜淼,等.麦门冬汤合苇茎汤抑制小鼠Lewis肺癌生长的体内实验研究[J].南京中医药大学学报,2011,27(2):144-147.
[9]王金辉,高然,明超,等.长链非编码RNA在非小细胞肺癌中的研究进展[J].现代肿瘤医学,2018,26(17):2819-2822.
[10]樊黎明,胡志东.LncRNA在非小细胞肺癌中的研究进展[J].中国肺癌杂志,2016,19(2):108-112.
[11]马雪曼,王笑民,于明薇,等.小鼠lewis肺癌原位移植瘤和异位皮下移植瘤模型的对比研究[J].实用癌症杂志,2018,33(4):523-526.
[12]姜雪莲,姚逸临,罗斌,等.低剂量化疗联合中医药治疗肺癌[J].长春中医药大学学报,2018,34(2):381-383.
[13]杨金坤.现代中医肿瘤学[M].上海:上海中医药大学出版社,2004:365.
[14]李炳照,陈海霞,李丽萍,等.实用中医方剂双解与临床[M].北京:科学技术文献出版社,2008:508.
[15]连建伟.方剂学[M].杭州:浙江科学技术出版社,2005:113.
[16]Reddy K B.MicroRNA(miRNA)in cancer[J].Cancer Cell Int,2015,15(1):1-6.
[17]YANG G,LU X,YUAN L.LncRNA:a link between RNA and cancer[J].Biochim Biophys Acta,2014,1839(11):1097-1109.
[18]NIE W,GE H J,YANG X Q,et al.LncRNA-UCA1exerts oncogenic functions in non-small cell lung cancer by targeting miR-193a-3p[J].Cancer Lett,2016,371(1):99-106.
[19]TAO H,YANG J J,ZHOU X,et al.Emerging role o long noncoding RNAs in lung cancer:current status and future prospects[J].Respir Med,2016,110(1):12-19.
[20]YANG Y,LI H,HOU S,et al.The noncoding RNAexpression profile and the effect of lncRNA AK126698on cisplatin resistance in non-small-cell lung cancer cel[J].PLo S One,2013,8(5):1-12.
[21]TANG Q,NI Z,CHENG Z,et al.Three circulating long non-coding RNAs act as biomarkers for predicting NSCLC[J].Cell Physiol Biochem,2015,37(3):1002-1009.
[22]郑璐玉,熊飞,詹7),等.麦门冬汤合千金苇茎汤提取部位对非小细胞肺癌H460细胞毒作用的研究[J].中国实验方剂学杂志,2010,16(3):60-63.
[23]施文,李俊生.ADAM家族与肿瘤关系的研究现状[J].实用癌症杂志,2008,23(6):671-673.
[24]ZHOU B B,Fridman J S,LIU X,et al.ADAMproteases,Erb B pathways and cancer[J].Expert Opin Investig Drugs,2005,14(6):591-606.
[25]Groner B,Von M V.Jak Stat signaling and cancer:Opportunities,benefits and side effects of targeted inhibition[J].Mol Cell Endocrinol,2017,451(1):1-14.
[26]Harada D,Takigawa N,Kiura K.The role of STAT3 in non-small cell lung cancer[J].Cancers,2014,6(2):708-722.
[27]张宏方,吴芳,环诚,等.调衡方对Lewis肺癌细胞增殖及JAK-STAT信号通路的影响[J].辽宁中医药大学学报,2013,15(3):16-19.