CN112946280A - Tgfbi基因在预测结直肠癌患者5-fu化疗反应中的应用 - Google Patents

Tgfbi基因在预测结直肠癌患者5-fu化疗反应中的应用 Download PDF

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CN112946280A
CN112946280A CN202110311336.XA CN202110311336A CN112946280A CN 112946280 A CN112946280 A CN 112946280A CN 202110311336 A CN202110311336 A CN 202110311336A CN 112946280 A CN112946280 A CN 112946280A
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colorectal cancer
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付凯
王雅
姚元炳
陈宇乔
魏群惠
龙仕超
林依琳
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Abstract

本发明属于基因的功能与应用领域，公开了TGFBI基因在预测结直肠癌患者5‑FU化疗反应中的应用，所述的TGFBI基因编码的蛋白质为SEQ ID NO.2所示。申请人发现TGFBI在HCT116、DLD1获得性耐药细胞中的表达均显著低于其亲本细胞株；在亲本细胞系HCT116、DLD1中用siRNA沉默TGFBI的表达后，细胞对5‑FU的敏感性降低；在HCT116获得性耐药细胞中分别过表达GFP、GFP‑TGFBI，发现过表达GFP‑TGFBI可以显著恢复获得性耐药HCT116细胞对5‑FU的敏感性。TGFBI高表达的结直肠癌患者可能在5‑FU治疗中获得更好的药物反应与预后。

Description

TGFBI基因在预测结直肠癌患者5-FU化疗反应中的应用
技术领域
本发明属于基因的功能与应用领域，具体涉及TGFBI基因在预测结直肠癌患者5-FU化疗反应中的应用，是以TGFBI基因作为预测结直肠癌患者对以5-FU为基础的化疗反应的生物标志物。
背景技术
结直肠癌是最常见的消化道肿瘤之一，III期结直肠癌患者和部分高风险II期结直肠癌患者通常会接受以5-FU为基础的化疗，尽管一部分患者会从化疗中获益，但也有很多患者可能会经历几个周期的无效化疗，直到治疗效果确定，这通常会带来不利的甚至危及生命的副作用。5-FU耐药是导致化疗失败的主要原因，而大约1/2患者对5-FU耐药。因此，迫切需要寻找新的生物标志物来预测5-FU化疗的反应，从而指导患者是否接受5-FU化疗。
目前一些预测结直肠癌患者基于5-FU治疗反应的生物标志物已经被报道：由TYMS基因编码的胸腺酸合成酶(thymidylate synthase,TS)的低表达，与结直肠癌患者对基于5-FU治疗的敏感性增加相关(1,2)。研究表明，DPYD基因编码的二氢吡啶脱氢酶(DPD)的表达是5-FU治疗有效性和毒性的预测标志物，肿瘤组织中高DPD活性可能通过降低5-FU的细胞毒性作用从而增加组织耐药性(3)。此外，DPD水平影响5-FU的分解代谢，低水平的DPD通过减少5-FU分解代谢导致药物在细胞内的有效积累(4)。据报道，DPYD在对5-FU无反应的结直肠癌组织中的表达比有反应的结直肠癌组织更高(5)。由TYMP基因编码的胸腺嘧啶磷酸化酶(TP)被发现是预测5-FU化疗有效性的有效标志物(6)。TP的低表达与改善的治疗结果具有相关性，TP的低表达预示着对5-FU化疗的良好反应(7,8)。然而，有研究得到了相反的结论：TP高表达的细胞可能与5-FU敏感性增高有关(3)。除了上述蛋白，膜转运蛋白也参与了5-FU化疗耐药机制，膜转运蛋白通过将药物转运出细胞，导致化疗失败。ATP结合盒式转运蛋白(ATP-binding cassette transporters)属于膜转运蛋白，一些ATP结合盒式转运蛋白也被报道跟结直肠癌患者5-FU耐药相关，例如ABCB5(9)、ABCC11(10)。虽然与5-FU耐药相关的蛋白及其机制已被报道，但应用于临床真正指导结直肠癌患者是否该接受5-FU化疗的生物标志物还没有。更多的5-FU耐药的生物标志物及其相关机制仍有待进一步研究。
TGFBI(transforming growth factorβ-induced protein,转化生长因子β诱导蛋白)，由TGFBI基因编码。TGFBI在TGFB(转化生长因子β)处理的人肺腺癌细胞株A549中首次被发现，它包含一个RGD(Arg-Gly-Asp)基序，该基序可以作为整合素的配体识别位点(11)。TGFBI通过结合整合素介导细胞与胶原蛋白、纤维连接蛋白和层粘连蛋白等细胞外蛋白的粘附(12)。有研究表明TGFBI具有抑癌作用，TGFBI的表达在多种肿瘤中均下调。免疫组织化学结果显示TGFBI在肺癌组织中的表达低于正常组织(13)。在卵巢癌组织中，启动子高甲基化导致TGFBI水平下调(14)。TGFBI启动子高甲基化也发生在肺癌和前列腺癌标本中(15)。在中国仓鼠卵巢(CHO)细胞中过表达TGFBI导致裸鼠体内CHO细胞生长和成瘤能力显著下降(16)。TGFBI的表达也降低了肺癌和乳腺癌细胞的转移潜能(17)。TGFBI在正常的羧基端加工过程中通过释放RGD多肽促进TGFBI诱导的凋亡(18)。TGFBI缺乏易使小鼠发生自发性肿瘤(19)。并且，在缺乏内源性TGFBI蛋白的肺癌细胞H522中，恢复TG FBI表达后，细胞生长显著下降，对凋亡的敏感性显著提高(13)。以上研究认为TGFBI发挥肿瘤抑制因子的作用。然而，TGFBI在肿瘤发展中的作用引起了争议，多项研究报道了TG FBI的促肿瘤功能。TGFBI增加了卵巢癌细胞的转移潜能，TGFBI可能是治疗卵巢癌的潜在药物靶点(20)。有研究表明TGFBI在卵巢癌中具有双重作用，根据肿瘤微环境的不同，TGFBI既可以作为肿瘤抑制因子，也可以作为肿瘤促进因子(21)。还有研究表明TGFBI可能发挥促肿瘤或抗肿瘤的作用，这取决于它结合的细胞表面的整合素类型(22)。
本研究首次发现TGFBI可能作为预测结直肠癌5-FU治疗反应的生物标志物。
参考文献：
1.Iyevleva AG,Buslov KG,Togo AV,Matsko DE,Filimonenko VP,MoiseyenkoVM,et al.Me asurement of DPD and TS transcripts aimed to predict clinicalbenefit from fluoropyrimidines:confirmation of the trend in Russiancolorectal cancer series and caution regarding the genereferees.Onkologie(2007)30(6):295-300.doi:10.1159/000102046.PubMed PMID:17551252.
2.Qiu LX,Tang QY,Bai JL,Qian XP,Li RT,Liu BR,et al.Predictive valueof thymidylate syn thase expression in advanced colorectal cancer patientsreceiving fluoropyrimidine-based chem otherapy:evidence from24studies.International journal of cancer(2008)123(10):2384-9.doi:10.1002/ijc.23822.PubMed PMID:18729195.
3.Panczyk M.Pharmacogenetics research on chemotherapy resistance incolorectal cancer over the last 20years.World journal of gastroenterology(2014)20(29):9775-827.doi:10.3748/wjg.v20.i29.9775.PubMed PMID:25110414；PubMed Central PMCID:PMC4123365.
4.Kunicka T,Prochazka P,Krus I,Bendova P,Protivova M,Susova S,etal.Molecular profil e of 5-fluorouracil pathway genes in colorectalcarcinoma.BMC cancer(2016)16(1):795.doi:10.1186/s12885-016-2826-8.PubMedPMID:27733154；PubMed Central PMCID:PMC5062913.
5.Salonga D,Danenberg KD,Johnson M,Metzger R,Groshen S,Tsao-Wei DD,etal.Colore ctal tumors responding to 5-fluorouracil have low gene expressionlevels of dihydropyrimidine dehydrogenase,thymidylate synthase,and thymidinephosphorylase.Clinical cancer research:an official journal of the AmericanAssociation for Cancer Research(2000)6(4):1322-7.PubMed PMID:10778957.
6.Meropol NJ,Gold PJ,Diasio RB,Andria M,Dhami M,Godfrey T,etal.Thymidine phosph orylase expression is associated with response tocapecitabine plus irinotecan in patients with metastatic colorectalcancer.Journal of clinical oncology:official journal of the American So cietyof Clinical Oncology(2006)24(25):4069-77.doi:10.1200/JCO.2005.05.2084.PubMedPMID:16943524.
7.Soong R,Shah N,Salto-Tellez M,Tai BC,Soo RA,Han HC,et al.Prognosticsignificance of thymidylate synthase,dihydropyrimidine dehydrogenase andthymidine phosphorylase protein expression in colorectal cancer patientstreated with or without 5-fluorouracil-based chemot herapy.Annals ofoncology:official journal of the European Society for Medical Oncology(2008)19(5):915-9.Epub 2008/02/05.doi:10.1093/annonc/mdm599.PubMed PMID:18245778；PubMed Central PMCID:PMCPmc2931808.
8.Gustavsson B,Kaiser C,Carlsson G,Wettergren Y,Odin E,Lindskog EB,etal.Molecular determinants of efficacy for 5-FU-based treatments in advancedcolorectal cancer:mRNA expression for 18 chemotherapy-relatedgenes.International journal of cancer(2009)124(5):1220-6.doi:10.1002/ijc.23852.PubMed PMID:19051292.
9.Wilson BJ,Schatton T,Zhan Q,Gasser M,Ma J,Saab KR,et al.ABCB5identifies a ther apy-refractory tumor cell population in colorectal cancerpatients.Cancer research(2011)71(15):5307-16.Epub 2011/06/10.doi:10.1158/0008-5472.can-11-0221.PubMed PMID:21652540；PubMed Central PMCID:PMCPmc3395026.
10.Hlavata I,Mohelnikova-Duchonova B,Vaclavikova R,Liska V,Pitule P,Novak P,et al.The role of ABC transporters in progression and clinicaloutcome of colorectal cancer.Mutagenesi s(2012)27(2):187-96.Epub 2012/02/02.doi:10.1093/mutage/ger075.PubMed PMID:22294766.
11.Skonier J,Neubauer M,Madisen L,Bennett K,Plowman GD,PurchioAF.cDNA cloning and sequence analysis of beta ig-h3,a novel gene induced in ahuman adenocarcinoma cell line after treatment with transforming growthfactor-beta.DNA and cell biology(1992)11(7):511-22.Epub 1992/09/01.doi:10.1089/dna.1992.11.511.PubMed PMID:1388724.
12.Kim JE,Kim SJ,Lee BH,Park RW,Kim KS,Kim IS.Identification ofmotifs for cell adhesi on within the repeated domains of transforming growthfactor-beta-induced gene,betaig-h3.The Journal of biological chemistry(2000)275(40):30907-15.Epub 2000/07/25.doi:10.1074/jbc.M002752200.PubMed PMID:10906123.
13.Zhao Y,El-Gabry M,Hei TK.Loss of Betaig-h3 protein is frequent inprimary lung carcin oma and related to tumorigenic phenotype in lung cancercells.Molecular carcinogenesis(2006)45(2):84-92.Epub 2005/12/06.doi:10.1002/mc.20167.PubMed PMID:16329146.
14.Kang S,Dong SM,Park NH.Frequent promoter hypermethylation of TGFBIin epithelial ovarian cancer.Gynecologic oncology(2010)118(1):58-63.Epub2010/04/28.doi:10.1016/j.ygyno.2010.03.025.PubMed PMID:20421128.
15.Shah JN,Shao G,Hei TK,Zhao Y.Methylation screening of the TGFBIpromoter in huma n lung and prostate cancer by methylation-specific PCR.BMCcancer(2008)8:284.Epub 2008/10/07.doi:10.1186/1471-2407-8-284.PubMed PMID:18834524；PubMed Central PMCID:PM CPmc2572632.
16.Skonier J,Bennett K,Rothwell V,Kosowski S,Plowman G,Wallace P,etal.beta ig-h3:a transforming growth factor-beta-responsive gene encoding asecreted protein that inhibits cell attachment in vitro and suppresses thegrowth of CHO cells in nude mice.DNA and cell biology(1994)13(6):571-84.Epub1994/06/01.doi:10.1089/dna.1994.13.571.PubMed PMID:8024701.
17.Wen G,Partridge MA,Li B,Hong M,Liao W,Cheng SK,et al.TGFBIexpression reduces in vitro and in vivo metastatic potential of lung andbreast tumor cells.Cancer Lett(2011)308(1):23-32.Epub 2011/05/13.doi:10.1016/j.canlet.2011.04.010.PubMed PMID:21561707；PubMed Central PMCID:PMCPmc3167173.
18.Kim JE,Kim SJ,Jeong HW,Lee BH,Choi JY,Park RW,et al.RGD peptidesreleased from beta ig-h3,a TGF-beta-induced cell-adhesive molecule,mediateapoptosis.Oncogene(2003)22(13):2045-53.Epub 2003/04/04.doi:10.1038/sj.onc.1206269.PubMed PMID:12673209.
19.Zhang Y,Wen G,Shao G,Wang C,Lin C,Fang H,et al.TGFBI deficiencypredisposes mi ce to spontaneous tumor development.Cancer research(2009)69(1):37-44.Epub 2009/01/02.doi:10.1158/0008-5472.can-08-1648.PubMed PMID:19117985；PubMed Central PMCID:PMC Pmc2664305.
20.Ween MP,Lokman NA,Hoffmann P,Rodgers RJ,Ricciardelli C,OehlerMK.Transforming growth factor-beta-induced protein secreted by peritonealcells increases the metastatic potent ial of ovarian cancercells.International journal of cancer(2011)128(7):1570-84.Epub 2010/06/04.doi:10.1002/ijc.25494.PubMed PMID:20521251.
21.Ween MP,Oehler MK,Ricciardelli C.Transforming growth Factor-Beta-Induced Protein(TG FBI)/(βig-H3):a matrix protein with dual functions inovarian cancer.International journal of molecular sciences(2012)13(8):10461-77.Epub 2012/09/06.doi:10.3390/ijms130810461.Pub Med PMID:22949874；PubMedCentral PMCID:PMCPmc3431872.
22.Thapa N,Lee BH,Kim IS.TGFBIp/betaig-h3 protein:a versatile matrixmolecule induced by TGF-beta.The international journal of biochemistry&cellbiology(2007)39(12):2183-94.Epub 2007/07/31.doi:10.1016/j.biocel.2007.06.004.PubMed PMID:17659994.
发明内容
本发明的目的在于提供了TGFBI基因在预测结直肠癌患者5-FU化疗反应中的应用，包括检测是以TGFBI基因作为预测结直肠癌患者对以5-FU为基础的化疗反应的生物标志物。
为了达到上述目的，本发明采取以下技术措施：
TGFBI基因在预测结直肠癌患者5-FU化疗反应中的应用，包括检测TGFBI基因表达量的物质或检测TGFBI蛋白水平的物质在制备预测结直肠癌患者5-FU化疗反应试剂中的应用，所述的TGFBI基因GenBank:BC000097.2；TGFBI基因表达量高，说明结直肠癌细胞对5-FU具有高敏感性，5-FU适合作为其化疗药物；若随着化疗时间增长，与初期相比，TGFBI基因表达量降低，说明5-FU不再适合作为化疗药物，需要变更治疗手段。
本发明的保护范围还包括：
包含有TGFBI基因的表达载体制备5-FU敏感性的结直肠癌细胞。
敲减TGFBI基因用于制备耐5-FU的结直肠癌细胞。
以上所述的细胞均可用于制备结直肠癌药物筛选模型。
本发明相对于现有技术具有如下的优点及效果：
(1)本发明首次发现了TGFBI的新功能：TGFBI可能作为预测结直肠癌患者5-FU化疗反应的生物标志物。申请人在结直肠癌细胞系中验证了其介导5-FU耐药性的功能。申请人选择了对5-FU相对敏感的结直肠癌细胞系HCT116、DLD1，通过浓度不断升高的5-FU持续处理细胞120天，获得了HCT116、DLD1的5-FU耐药细胞株，通过Western blot检测TGFBI在亲本和获得性耐药细胞中的表达，发现TGFBI在HCT116、DLD1获得性耐药细胞中的表达均显著低于其亲本细胞株；申请人在亲本细胞系HCT116、DLD1中用siRNA敲减TGFBI的表达后，通过MTT实验检测细胞对5-FU的敏感性，结果显示：在HCT116、DL D1细胞中敲减TGFBI后，细胞对5-FU的敏感性降低；申请人又进一步在HCT116获得性耐药细胞中分别过表达GFP、GFP-TGFBI，发现过表达GFP-TGFBI可以显著恢复获得性耐药HCT116细胞对5-FU的敏感性。因此，TGFBI的高表达预示着细胞对5-FU处理的高敏感性，TGFBI高表达的结直肠癌患者可能在5-FU治疗中获得更好的药物反应与预后。
(2)对TCGA(the cancer genome atlas)数据库中结直肠癌化疗患者的生存进行分析，其中TGFBI的表达与结直肠癌化疗患者的生存显著相关，且TGFBI高表达患者的生存期显著高于TGFBI低表达患者。为了进一步验证TGFBI水平与5-FU敏感性的关系，我们对GEO数据集GSE19860中结直肠癌患者样本也进行了分析，发现接受以5-FU为基础的化疗患者中，对化疗应答患者中TGFBI的表达显著高于化疗无应答患者。因此，TGFBI高表达的结直肠癌患者对5-FU治疗更敏感。TGFBI可能作为预测化疗疗效的生物标志物。
附图说明
图1为亲本细胞和5-FU耐药细胞MTT实验结果示意图；
其中：A为HCT116的亲本细胞和耐药细胞的MTT实验结果，HCT116-P为HCT116亲本细胞，HCT116-R为HCT116耐5-FU细胞；
B为DLD1细胞的亲本细胞和耐药细胞的MTT实验结果，DLD1-P为DLD1亲本细胞，DLD1-R为DLD1耐5-FU细胞。图中*表示p<0.05。
图2为利用Western blot检测耐5-FU细胞和亲本细胞中TGFBI的表达情况的示意图；
图中“+”表示该泳道对应的样品。
图3为亲本细胞敲减TGFBI基因后对5-FU的敏感性检测示意图；
“+”表示该泳道对应的样品；
其中：A为Western blot检测细胞内的TGFBI敲减情况；si-NC表示以乱序RNA转染细胞，si-TGFBI表示以特异性干扰TGFBI基因的小RNA转染细胞；结果显示转染特异性干扰TGFBI基因的小RNA可以成功干扰TGFBI在HCT116细胞和DLD1细胞中的表达；
B的左图为HCT116乱序RNA对照组细胞和TGFBI干扰组细胞对5-FU的敏感性实验结果示意图，si-NC表示以乱序RNA转染细胞，si-TGFBI表示以特异性干扰TGFBI基因的小RNA转染细胞；
B的右图为DLD1乱序RNA对照组细胞和TGFBI干扰组细胞对5-FU的敏感性实验结果示意图，si-NC表示以乱序RNA转染细胞，si-TGFBI表示以特异性干扰TGFBI基因的小RNA转染细胞。图中*表示p<0.05。
图4为不同细胞转染不同的质粒后对5-FU的敏感性实验结果示意图；
“+”表示该泳道对应的样品；
其中A为Western blot检测不同细胞转染不同的质粒后表达GFP或者TGFBI的效率；
B为细胞过表达GFP或者TGFBI后，加入不同浓度5-FU处理，MTT实验检测细胞活力示意图。图中*表示p<0.05；ns表示p>0.05，无显著差异。
图5为TCGA数据库中结直肠癌化疗患者TGFBI高表达组和TGFBI低表达组的五年生存曲线示意图。
图6为GEO数据集GSE19860中结直肠癌化疗敏感组患者和化疗耐受组患者中TGFBI的表达量示意图。
具体实施方式
通过以下详细说明结合附图可以进一步理解本发明的特点和优点。所提供的实施例仅是对本发明方法的说明，而不以任何方式限制本发明揭示的其余内容。本发明所述技术方案，如未特别说明，均为本领域的常规技术；所述试剂或材料，如未特别说明，均来源于商业渠道。
实施例1：
TGFBI基因影响结直肠癌细胞对5-FU的耐药性：
本发明所述的TGFBI基因GenBank:BC000097.2，其编码的蛋白质为SEQ ID NO.2所示。本发明实施例以购买的质粒为模板扩增的TGFBI基因，其CDS序列为SEQ ID NO.1所示，碱基序列跟NCBI公布的相比，有同义突变的碱基，但氨基酸序列与NCBI上公开的完全相同。
本发明实施例所用的细胞为对5-FU相对敏感的结直肠癌细胞系HCT116、DLD1，也称为亲本细胞。
1)5-FU获得性耐药细胞系的制备及耐药验证：
本发明中细胞所用培养基均为DMEM+10％胎牛血清+青霉素100单位/ml+链霉素0.1mg/ml，培养环境均为37℃，5％CO2。
5-FU获得性耐药细胞系HCT116：HCT116细胞用40μM5-FU处理24h后，更换培养基，用0.3125μM5-FU继续处理，之后按1.5倍浓度增加5-FU浓度，维持1.5倍5-FU处理细胞两周，每隔两周将5-FU浓度增至上一次处理浓度的1.5倍，处理4个月后，得到5-FU获得性耐药细胞系HCT116。
5-FU获得性耐药细胞系DLD1：DLD1细胞用100μM5-FU处理24h后，更换培养基，用1.25μM5-FU继续处理两周，之后按1.5倍浓度增加5-FU浓度，维持1.5倍5-FU处理细胞两周，每隔两周将5-FU浓度增至上一次处理浓度的1.5倍，处理4个月后，得到5-FU获得性耐药细胞系DLD1。
分别对亲本细胞HCT116、DLD1及其上述制备的耐药细胞进行了5-FU IC50的测定。亲本细胞和耐药细胞置于96孔板中，待细胞贴壁后，在细胞中加入含不同浓度5-FU(0.2μM、2μM、10μM、100μM)的培养基，不同浓度5-FU处理的细胞设置3个重复；5-FU处理72h后，弃掉培养基，加入含MTT的培养基100μL(每100μL培养基含有5μL MTT(5mg/ml))，37℃孵育4h后，弃掉培养基，加入100μL DMSO，摇床上震荡10min后，酶标仪检测OD490的吸光度值，并计算细胞5-FU IC50值。
结果如图1所示：相同浓度5-FU处理时，HCT116耐药细胞株和DLD1耐药细胞株的细胞活力均显著高于相应的亲本细胞系，对IC50值进行计算，亲本HCT116细胞、5-FU耐药HCT116细胞、亲本DLD1细胞、5-FU耐药DLD1细胞的IC50值分别为4.032μM、40.085μM、2.091μM、70.820μM。结果表明：耐药细胞系筛选成功。
2)耐药细胞和亲本细胞中TGFBI的表达量的检测：
分别接种HCT116和DLD1的亲本细胞和耐药细胞于12孔板中，待细胞贴壁生长24h后，用胰酶消化细胞，收集细胞，加入适量的细胞裂解液，于冰上裂解细胞，之后加入等量的2×loading buffer，用于Western blot检测。TGFBI抗体和Actin抗体分别用于检测细胞中TGF BI和Actin的表达情况，其中Actin是内参。
结果如图2所示：细胞中Actin的量一致，表示点样量一致，而5-FU耐药HCT116细胞中TGFBI的量显著低于亲本HCT116细胞，5-FU耐药DLD1细胞中TGFBI的量也显著低于亲本DLD1细胞。结果表明：耐药细胞中TGFBI的表达量显著低于亲本敏感细胞。
3)利用siRNA将亲本细胞中的TGFBI沉默后，检测细胞对5-FU敏感性：
分别接种亲本HCT116、DLD1细胞于12孔板中，待细胞贴壁生长12h后进行转染。转染按以下步骤进行：在100μL Opti-MEM中加入40pmol siRNA,柔和混匀；在100μL Opti-MEM中加入0.5μL lipofectamine2000,轻轻混匀，室温放置5min；将稀释好的siRNA和lipofectam in2000试剂混合，轻轻混匀后室温放置20min；将200μL siRNA/lipofectamin复合物加入到含有细胞的培养板的孔中。
实验分为四组：①HCT116细胞转染乱序RNA组，即si-NC；②HCT116细胞转染TGFBI特异性小RNA组，即si-TGFBI；③DLD1细胞转染乱序RNA组，即si-NC；④DLD1细胞转染TGFBI特异性小RNA组，即si-TGFBI。转染72h后，消化细胞，一半用于Westernblot检测基因沉默效率，另外一半用于MTT实验检测沉默TGFBI后细胞对5-FU的敏感性是否发生变化。
Western blot实验中，TGFBI抗体和Actin抗体分别用来检测细胞中TGFBI和Actin的量。结果如图3中A所示：HCT116细胞中，si-NC组和si-TGFBI组的Actin的量一致，表明细胞上样量一致，而si-TGFBI组中TGFBI的量明显低于si-NC组，表明HCT116细胞中TGFBI的表达被有效沉默；与HCT116细胞一致，DLD1细胞中TGFBI的表达也被有效沉默。
HCT116和DLD1细胞分别经过乱序对照RNA(si-NC)和TGFBI特异性小RNA(si-TGFBI)转染72h后，进行MTT实验，实验结果如图3中B所示：在HCT116细胞中，在0.4μM、1.6μM、6.4μM、25.6μM5-FU浓度处理时，si-NC组细胞存活率分别为84.3％、75.7％、45.8％和24.5％，而si-TGFBI组细胞存活率分别为98.3％、102.9％、80.5％和40.4％，在相同浓度5-FU处理下，si-TGFBI组细胞存活更多，表明在HCT116细胞中沉默TGFBI的表达后，细胞对5-FU处理更耐受；在DLD1细胞中，在1.6μM、6.4μM、25.6μM、102.4μM 5-FU浓度处理时，si-NC组细胞的存活率分别为：52.4％、17.4％、14.2％、10.9％，而si-TGFBI组细胞的存活率分别为：65.6％、42.0％、34.9％、29.6％，在相同浓度5-FU处理下，si-TGFBI组细胞存活更多，表明在DLD1细胞中沉默TGFBI的表达后，细胞对5-FU处理更耐受。
4)在耐5-FU HCT116细胞中过表达TGFBI后检测细胞对5-FU的敏感性：
(1)pEGFP-C1-TGFBI表达载体的构建：
以pOTB7-TGFBI(P14682)(购自淼灵质粒基因平台，货号是P14682)为模板，利用引物5’-GATCTCGAGCTCAAGCTTCGAATTCCATGGCGCTCTTCGTGCGG-3’和5’-GATCTCGAGCTCAAGCTTCGAATTCCATGGCGCTCTTCGTGCGGTCAGTTATCTAGATCCGGTGGATCCCTAATGCTTCATCCTCTCTAATAACTTTTGATAGACAG-3’，扩增全长的TGFBI。质粒pEGFP-C1通过EcoRⅠ、BamHⅠ双酶切进行线性化；随后采用Gibson组装的方式将TGFBI构建入pEGFP-C1载体中，构建的载体经测序鉴定正确，即得pEGFP-C1-TGFBI表达载体。
(2)过表达TGFBI可以恢复耐5-FU HCT116细胞对5-FU的敏感性：
以步骤1)制备的5-FU获得性耐药细胞系HCT116(HCT116-R)为研究对象，接种HCT116-R细胞于12孔板的2个孔，分别用于转染pEGFP-C1和pEGFP-C1-TGFBI质粒，同时接种HCT116亲本敏感细胞(HCT116-P)于12孔板的1个孔，用于转染pEGFP-C1质粒做对照。待细胞贴壁生长12h后进行转染，转染采用lipofectamine2000试剂，转染过程与步骤3)中转染siRNA相似。转染24h后，消化细胞，收集细胞，一半用于Western blot检测转染效率，另一半接种96孔板，用不同浓度5-FU处理72h后，进行MTT实验，检测细胞对5-FU的敏感性。
Western blot实验分别用GFP抗体和Actin抗体检测细胞中GFP和Actin的表达情况，其中Actin作为内参，如图4中A所示：在转染pEGFP-C1的HCT116-P细胞和HCT116-R细胞中检测到大小为27KDa的GFP条带，而HCT116-R细胞中由于转染了pEGFP-C1-TGFBI质粒，在27KDa位置未检测到条带，在95KDa处检测到GFP条带(GFP蛋白为27KDa，TGFBI蛋白大小为68KDa)，表明HCT116-P细胞和HCT116-R细胞均成功表达转染质粒。
对成功转染pEGFP-C1和pEGFP-C1-TGFBI质粒的HCT116-P细胞和HCT116-R细胞用不同浓度5-FU处理72h后，进行MTT实验。结果如图4中B所示：在相同浓度5-FU处理下，过表达GFP的HCT116-R细胞的存活率显著高于过表达GFP的HCT116-P细胞的存活率。在50μM、150μM、500μM5-FU浓度处理时，过表达GFP-TGFBI的HCT116-R细胞的存活率分别为：40.8％、37.2％和15.7％，而过表达GFP的HCT116-R细胞的存活率分别为：66.4％、51.1％、48.2％，在HCT116-R细胞中过表达TGFBI后，细胞的存活率显著低于GFP对照组细胞，表明在HCT116-R细胞中过表达TGFBI后恢复了细胞对5-FU的敏感性。
实施例2：
生物信息学分析验证TGFBI基因与结直肠癌患者5-FU化疗敏感性显著相关：
为了确认TGFBI基因的表达确实跟结直肠癌患者5-FU化疗敏感性相关，我们对TCGA(the cancer genome atlas)数据库中结直肠癌化疗患者的生存进行分析，其中TGFBI基因的表达与结直肠癌化疗患者的生存显著相关，且TGFBI高表达患者的生存期显著高于TGFBI低表达患者(图5)。为了进一步验证TGFBI水平与5-FU敏感性的关系，我们对GEO数据集GSE19860中结直肠癌患者样本也进行了分析，发现接受以5-FU为基础的化疗患者中，对化疗敏感的患者中TGFBI的表达显著高于化疗耐受患者(图6)。因此，TGFBI高表达的结直肠癌患者对5-FU治疗更敏感。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。
序列表
<110> 中南大学湘雅医院
<120> TGFBI基因在预测结直肠癌患者5-FU化疗反应中的应用
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Val Gly Ser Thr Thr Thr Gln Leu Tyr Thr Asp Arg Thr Glu Lys Leu
115 120 125
Arg Pro Glu Met Glu Gly Pro Gly Ser Phe Thr Ile Phe Ala Pro Ser
130 135 140
Asn Glu Ala Trp Ala Ser Leu Pro Ala Glu Val Leu Asp Ser Leu Val
145 150 155 160
Ser Asn Val Asn Ile Glu Leu Leu Asn Ala Leu Arg Tyr His Met Val
165 170 175
Gly Arg Arg Val Leu Thr Asp Glu Leu Lys His Gly Met Thr Leu Thr
180 185 190
Ser Met Tyr Gln Asn Ser Asn Ile Gln Ile His His Tyr Pro Asn Gly
195 200 205
Ile Val Thr Val Asn Cys Ala Arg Leu Leu Lys Ala Asp His His Ala
210 215 220
Thr Asn Gly Val Val His Leu Ile Asp Lys Val Ile Ser Thr Ile Thr
225 230 235 240
Asn Asn Ile Gln Gln Ile Ile Glu Ile Glu Asp Thr Phe Glu Thr Leu
245 250 255
Arg Ala Ala Val Ala Ala Ser Gly Leu Asn Thr Met Leu Glu Gly Asn
260 265 270
Gly Gln Tyr Thr Leu Leu Ala Pro Thr Asn Glu Ala Phe Glu Lys Ile
275 280 285
Pro Ser Glu Thr Leu Asn Arg Ile Leu Gly Asp Pro Glu Ala Leu Arg
290 295 300
Asp Leu Leu Asn Asn His Ile Leu Lys Ser Ala Met Cys Ala Glu Ala
305 310 315 320
Ile Val Ala Gly Leu Ser Val Glu Thr Leu Glu Gly Thr Thr Leu Glu
325 330 335
Val Gly Cys Ser Gly Asp Met Leu Thr Ile Asn Gly Lys Ala Ile Ile
340 345 350
Ser Asn Lys Asp Ile Leu Ala Thr Asn Gly Val Ile His Tyr Ile Asp
355 360 365
Glu Leu Leu Ile Pro Asp Ser Ala Lys Thr Leu Phe Glu Leu Ala Ala
370 375 380
Glu Ser Asp Val Ser Thr Ala Ile Asp Leu Phe Arg Gln Ala Gly Leu
385 390 395 400
Gly Asn His Leu Ser Gly Ser Glu Arg Leu Thr Leu Leu Ala Pro Leu
405 410 415
Asn Ser Val Phe Lys Asp Gly Thr Pro Pro Ile Asp Ala His Thr Arg
420 425 430
Asn Leu Leu Arg Asn His Ile Ile Lys Asp Gln Leu Ala Ser Lys Tyr
435 440 445
Leu Tyr His Gly Gln Thr Leu Glu Thr Leu Gly Gly Lys Lys Leu Arg
450 455 460
Val Phe Val Tyr Arg Asn Ser Leu Cys Ile Glu Asn Ser Cys Ile Ala
465 470 475 480
Ala His Asp Lys Arg Gly Arg Tyr Gly Thr Leu Phe Thr Met Asp Arg
485 490 495
Val Leu Thr Pro Pro Met Gly Thr Val Met Asp Val Leu Lys Gly Asp
500 505 510
Asn Arg Phe Ser Met Leu Val Ala Ala Ile Gln Ser Ala Gly Leu Thr
515 520 525
Glu Thr Leu Asn Arg Glu Gly Val Tyr Thr Val Phe Ala Pro Thr Asn
530 535 540
Glu Ala Phe Arg Ala Leu Pro Pro Arg Glu Arg Ser Arg Leu Leu Gly
545 550 555 560
Asp Ala Lys Glu Leu Ala Asn Ile Leu Lys Tyr His Ile Gly Asp Glu
565 570 575
Ile Leu Val Ser Gly Gly Ile Gly Ala Leu Val Arg Leu Lys Ser Leu
580 585 590
Gln Gly Asp Lys Leu Glu Val Ser Leu Lys Asn Asn Val Val Ser Val
595 600 605
Asn Lys Glu Pro Val Ala Glu Pro Asp Ile Met Ala Thr Asn Gly Val
610 615 620
Val His Val Ile Thr Asn Val Leu Gln Pro Pro Ala Asn Arg Pro Gln
625 630 635 640
Glu Arg Gly Asp Glu Leu Ala Asp Ser Ala Leu Glu Ile Phe Lys Gln
645 650 655
Ala Ser Ala Phe Ser Arg Ala Ser Gln Arg Ser Val Arg Leu Ala Pro
660 665 670
Val Tyr Gln Lys Leu Leu Glu Arg Met Lys His
675 680
<210> 3
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
gatctcgagc tcaagcttcg aattccatgg cgctcttcgt gcgg 44
<210> 4
<211> 107
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gatctcgagc tcaagcttcg aattccatgg cgctcttcgt gcggtcagtt atctagatcc 60
ggtggatccc taatgcttca tcctctctaa taacttttga tagacag 107

Claims (4)

1.检测TGFBI基因表达量或检测TGFBI蛋白水平的物质在制备预测结直肠癌患者5-FU化疗反应试剂中的应用，所述的TGFBI基因编码的蛋白质为SEQ ID NO.2所示。
2.包含有TGFBI基因的表达载体在制备5-FU敏感性的结直肠癌细胞中的应用，所述的TGFBI基因编码的蛋白质为SEQ ID NO.2所示。
3.敲减TGFBI基因用于制备耐5-FU的结直肠癌细胞，所述的TGFBI基因编码的蛋白质为SEQ ID NO.2所示。
4.权利要求2或3所述的细胞在制备结直肠癌药物筛选模型中的应用。
CN202110311336.XA 2021-03-24 2021-03-24 Tgfbi基因在预测结直肠癌患者5-fu化疗反应中的应用 Pending CN112946280A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113881707A (zh) * 2021-10-25 2022-01-04 中国人民解放军军事科学院军事医学研究院 调控脐带间充质干细胞免疫抑制作用的产品、方法及用途

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090258795A1 (en) * 2007-03-15 2009-10-15 Genomic Health, Inc. Gene expression markers for prediction of patient response to chemotherapy

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090258795A1 (en) * 2007-03-15 2009-10-15 Genomic Health, Inc. Gene expression markers for prediction of patient response to chemotherapy

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
AHMED ASHOUR AHMED等: "The Extracellular Matrix Protein TGFBI Induces Microtubule Stabilization and Sensitizes Ovarian Cancers to Paclitaxel", 《CANCER CELL》 *
ESTHER GRAUDENS等: "Deciphering cellular states of innate tumor drug responses", 《GENOME BIOLOGY》 *
IRIGOYEN MARTA等: "TGFBI expression is associated with a better response to chemotherapy in NSCLC", 《MOL CANCER》 *
MELANIE SPITZNER等: "A gene expression signature for chemoradiosensitivity of colorectal cancer cells", 《INT. J. RADIATION ONCOLOGY BIOL. PHYS.》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113881707A (zh) * 2021-10-25 2022-01-04 中国人民解放军军事科学院军事医学研究院 调控脐带间充质干细胞免疫抑制作用的产品、方法及用途

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