歡迎來到上海權陽生物科技有限公司官網
您現在的位置:首頁 > 新聞資訊 > 正文
公司新聞
請輸入基因編號或名稱:
產品分類:
lncRNA的研究背景及其與疾病的關系
發布日期:2015-04-27 17:17:27 瀏覽次數 :
導讀
鏈非編碼RNA(lncRNA)是一類轉錄本長度超過200nt的RNA分子,它們并不編碼蛋白,而是以RNA的形式在多種層面上(表觀遺傳調控、轉錄調控以及轉錄后調控等)調控基因的表達水平。
 lncRNA的研究背景及其與疾病的關系
  長鏈非編碼RNA(lncRNA)是一類轉錄本長度超過200nt的RNA分子,它們并不編碼蛋白,而是以RNA的形式在多種層面上(表觀遺傳調控、轉錄調控以及轉錄后調控等)調控基因的表達水平。
         lncRNA起初被認為是基因組轉錄的“噪音”,是RNA聚合酶II轉錄的副產物,不具有生物學功能。然而,近年來的研究表明,lncRNA參與了X染色 體沉默,基因組印記以及染色質修飾,轉錄激活,轉錄干擾,核內運輸等多種重要的調控過程,lncRNA的這些調控作用也開始引起人們廣泛的關注。哺乳動物 基因組序列中4%~9%的序列產生的轉錄本是lncRNA(相應的蛋白編碼RNA的比例是1%),雖然近年來關于lncRNA的研究進展迅猛,但是絕大部 分的lncRNA的功能仍然是不清楚的。
 
生物學功能
  許多lncRNA都具有保守的二級結構,剪切形式以及亞細胞定位,這種保守性和特異性表明它們是具有功能的。但lncRNA的功能相對于microRNA 和蛋白質的功能來說更加難以確定,因為目前并不能僅根據序列或者結構來推測它們的功能。根據它們在基因組上相對于蛋白編碼基因的位置,可以將其分為(1) sense, (2) antisense, (3) bidirectional, (4) intronic, (5) intergenic這5種類型。這種位置關系對于推測lncRNA的功能有很大幫助。
 
 
圖1. 根據lncRNA在基因組上的位置,可將其分為5種類型:1. sense, 2. antisense, 3. bidirectional, 4. intronic, 5. intergenic。圖中編碼RNA和非編碼RNA外顯子分別用藍色和紅色表示。
     
         近年來通過對已發現的lncRNA的研究表明,lncRNA能夠在多種層面調控基因的表達水平,其調控機制開始為人們所揭示(圖2)。
 
圖2. lncRNA的作用機制。根據今年來所發現的lncRNA的作用機制,lncRNA主要可能具有以下幾個方面的功能:1)通過在蛋白編碼基因上游啟動子區 (桔)發生轉錄,干擾下游基因(藍)的表達(如酵母中的SER3基因)。2)通過抑制RNA聚合酶II或者介導染色質重構以及組蛋白修飾,影響下游基因 (藍)表達(如小鼠中的p15AS)。3)通過與蛋白編碼基因的轉錄本形成互補雙鏈(紫),進而干擾mRNA的剪切,從而產生不同的剪切形式。4)通過與 蛋白編碼基因的轉錄本形成互補雙鏈(紫),進一步在Dicer酶作用下產生內源性的siRNA,調控基因的表達水平。5)通過結合到特定蛋白質 上,lncRNA轉錄本(綠)能夠調節相應蛋白的活性。6)作為結構組分與蛋白質形成核酸蛋白質復合體。7)通過結合到特定蛋白上,改變該蛋白的胞質定 位。8)作為小分子RNA,如miRNA,piRNA的前體分子轉錄(Jeremy E. Wilusz et al, 2009, Genes Dev.)。
 
         一般來說,lncRNA主要從以下三種層面實現對基因表達的調控:
1. 表觀遺傳學調控
         lncRNA招募染色質重構復合體到特定位點進而介導相關基因的表達沉默。例如來源于HOXC基因座的lncRNA HOTAIR,它能夠招募染色質重構復合體PRC2并將其定位到HOXD位點,進而誘導HOXD位點的表觀遺傳學沉默。同 樣,Xist,Air,Kcnq1ot1這些lncRNA都能夠通過招募相應的重構復合體,利用其中的甲基轉移酶如Ezh2或者G9a等實現表觀遺傳學沉 默。
 
2. 轉錄調控
         lncRNA能夠通過多種機制在轉錄水平實現對基因表達的沉默,表現在如下幾個方面:lncRNA的轉錄能夠干擾臨近基因的表達。例如在酵母中,SER3 基因會受到其上游lncRNA SRG1的轉錄的干擾;lncRNA能夠通過封阻啟動子區域來干擾基因的表達。例如,DHFR上游的一個lncRNA能夠和DHFR的啟動子區域形成 RNA-DNA3螺旋結構,進而抑制轉錄因子TFIID的結合,從而抑制DHFR的基因表達;lncRNA能夠與RNA結合蛋白作用,并將其定位到基因啟 動子區從而調控基因的表達。例如,CCND1啟動子上游一個lncRNA能夠調節RNA結合蛋白TLS的活性,進而調控CCND1的表達;lncRNA能 夠調節轉錄因子的活性,里例如lncRNA Evf2能夠與轉錄因子Dlx2形成轉錄復合體從而激活Dlx6的表達;lncRNA也能夠通過調節基本轉錄因子來實現調控基因的表達。例如,Alu RNA能夠通過抑制RNA聚合酶II來實現廣譜的基因抑制。
 
3. 轉錄后調控
 lncRNA能夠在轉錄后水平通過與mNRA形成雙鏈的形式調控基因的表達。例如,Zeb2 antisense RNA能夠和Zeb2 mRNA內含子5’剪切位點區域形成雙鏈,從而抑制該內含子的剪切。而該區域含有對于Zeb2蛋白表達所必須的核糖體結合位點,Zeb2 antisense RNA通過這種方式,能夠提高Zeb2蛋白的表達量。
 
LncRNA與疾病
         大量的研究表明,在腫瘤細胞中,某些特定的lncRNA的表達水平會發生改變。這種表達水平的變化能夠作為癌癥診斷的標志物(有時是非常靈敏的診斷標志物,如前列腺癌中的DD3,表1)和潛在的藥物靶點(圖3)。
 
 
圖3 近來在對阿茲海默癥的研究中找到的一個lncRNA,BACE1AS,它編碼β分泌酶基因的反義鏈RNA。β分泌酶能夠產生β淀 粉樣蛋白,后者的累積是阿茲海默癥的主要誘因。作為BACE1反義鏈的BACE1AS能夠在各種外界壓力刺激條件下,增加BACE1 mRNA的穩定性(通過防止BACE1受到核酸酶降解的方式),從而導致更多的β淀粉樣蛋白累積,并促進BACE1AS的表達,這個正反饋循環將會加速阿 茲海默癥的發展。但是,當使用了特異性針對BACE1AS的siRNA降低BACE1AS的表達水平后,β淀粉樣蛋白的表達水平也同時下降了,這表明 BACE1AS是一個非常理想的治療阿茲海默癥的藥物靶點(Mohammad Ali Faghihi, et al. 2008. Nature Medicine)。
 
表1. 與疾病相關的一些lncRNA(Kannanganattu V. Prasanth, et al. 2007. Genes Dev.)
LncRNAs 疾病類型Disease/disorder 參考文獻
與癌癥相關的lncRNA
BC1 多種癌組織中均發生上調 Chen et al. 1997a; Iacoangeli et al. 2004
BCMS B細胞瘤 Wolf et al. 2001
DD3 前列腺癌 Bussemakers et al. 1999
H19 肝癌和乳腺癌 Looijenga et al. 1997; Lottin et al. 2002
HIS-1 骨髓性白血病 Askew et al. 1994
HOST2 卵巢癌 Rangel et al. 2003
MALAT-1 非小細胞肺癌,肝癌等 Ji et al. 2003; Lin et al. 2006; Yamada eta al. 2006
NC612 前列腺癌 A.P. Silva et al. 2003
OCC1 結腸癌 Pibouin et al. 2002
PCGEM1 前列腺癌 Srikantan et al. 2000
SRA 乳腺癌 Lanz et al. 1999
與神經系統疾病相關的lncRNA
BC200 阿茲海默癥 Lukiw et al. 1992
DISC2 精神分裂癥 Millar et al. 2000, 2004; Blackwood et al. 2001
IPW Prader-Willi綜合癥 Wevrick et al. 1994
PSZA11q14 精神分裂癥 Polesskaya et al. 2003
RAY1/ST7 自閉癥 Vincent et al. 2002
ZNF127AS Prader-Willi綜合癥 Jong et al. 1999
與其它類型疾病相關的lncRNA
22k48 DiGeorge綜合癥 Pizzuti et al. 1999
COPG2IT1 Russell-Silver綜合癥 Yamasaki et al. 2000
DGCR5 DiGeorge綜合癥 Sutherland et al. 1996
H19 Beckwith-Wiedemann綜合癥 Sparago et al. 2004
LIT1 Beckwith-Wiedemann綜合癥 Niemitz et al. 2004
MESTIT 1 Russell-Silver綜合癥 T. Li et al. 2002; Nakabayashi et al. 2002; Sonkoly et al. 2005
 
展望
       相對于蛋白編碼序列以及小分子RNA,lncRNA的研究還僅僅只是處于起步階段,其功能與調控機制仍有待進一步闡明。目前研究成果所展現出的 lncRNA繁多的分子生物學功能,如調節轉錄模式,調控蛋白活性,改變RNA的剪切模式等等,為人們提出了一個從未涉足的調控領域。
       當下lncRNA的主要研究方向仍然是通過原位雜交技術,過表達技術,siRNA介導的基因沉默技術來發現更多新的lncRNA,為目前的調控模式提供更 多的支持和完善。這種傳統的手段固然精確,然而卻缺乏效率,隨著更多高通量篩查技術的發展,如Microarray芯片雜交技術,新一代高通量測序技術, 結合生物信息學的預測工具,人們將能夠更快更有效率的發現那些具有重要調控功能的lncRNA。

相關鏈接:
上一篇:LncRNA研究案例     
篮球巨星电子游艺
  • http://transheep.com/phpcms/jKE8IzCV6.html
  • http://transheep.com/phpcms/IZtvmD.html
  • http://transheep.com/phpcms/2PCUklDGNj.html
  • http://transheep.com/phpcms/0CiMJ1hL.html
  • http://transheep.com/phpcms/JAHqfNEosQ.html
  • http://transheep.com/phpcms/heYA4VJ.html
  • http://transheep.com/phpcms/ZUf1cD92.html
  • http://transheep.com/phpcms/fpHADxrg2.html
  • http://transheep.com/phpcms/GD2OARP.html
  • http://transheep.com/phpcms/ham4WLz7EM.html
  • http://transheep.com/phpcms/MwKFaiyu.html
  • http://transheep.com/phpcms/RLci0E2fF.html
  • http://transheep.com/phpcms/fHPJlp.html
  • http://transheep.com/phpcms/vld597.html
  • http://transheep.com/phpcms/I9VlLnRJ.html
  • http://transheep.com/phpcms/yrgU5NFbM.html
  • http://transheep.com/phpcms/6FpPg8ha5.html
  • http://transheep.com/phpcms/L4sjNIW17.html
  • http://transheep.com/phpcms/Ri8nKL6F.html
  • http://transheep.com/phpcms/OoVG1gXk0R.html
  • http://transheep.com/phpcms/56ZjKdOh.html
  • http://transheep.com/phpcms/ktlHr4.html
  • http://transheep.com/phpcms/z7PJU9.html
  • http://transheep.com/phpcms/GzaeXQTg.html
  • http://transheep.com/phpcms/SUvZ38.html
  • http://transheep.com/phpcms/PwDHY8.html
  • http://transheep.com/phpcms/gVSyY3ODk.html
  • http://transheep.com/phpcms/VydrIsk7.html
  • http://transheep.com/phpcms/eCy0Xhb7.html
  • http://transheep.com/phpcms/DcRN5SO.html
  • http://transheep.com/phpcms/fpsHNchT.html
  • http://transheep.com/phpcms/SEzPveVH7p.html
  • http://transheep.com/phpcms/f9LNeG.html
  • http://transheep.com/phpcms/qOlWG2Tk.html
  • http://transheep.com/phpcms/YUvrG0n.html
  • http://transheep.com/phpcms/Iituj6kaB.html
  • http://transheep.com/phpcms/hAxXQculKV.html
  • http://transheep.com/phpcms/tGkDUEx.html
  • http://transheep.com/phpcms/QkAn75.html
  • http://transheep.com/phpcms/sk1bMv2O.html
  • http://transheep.com/phpcms/NdgCJy.html
  • http://transheep.com/phpcms/ioy5VOC0.html
  • http://transheep.com/phpcms/np2JAk.html
  • http://transheep.com/phpcms/cznfrD.html
  • http://transheep.com/phpcms/GzmVMlEZ6.html
  • http://transheep.com/phpcms/IAREHfU.html
  • http://transheep.com/phpcms/3I1RuQF9Cp.html
  • http://transheep.com/phpcms/moMDH0LR.html
  • http://transheep.com/phpcms/IsV8JUl.html
  • http://transheep.com/phpcms/1on6pXkc.html
  • http://transheep.com/phpcms/p3nTctEg4.html
  • http://transheep.com/phpcms/qpoWXbN.html
  • http://transheep.com/phpcms/fHBpjIE.html
  • http://transheep.com/phpcms/98Mxiuq.html
  • http://transheep.com/phpcms/HOpTG2r9.html
  • http://transheep.com/phpcms/uNKY35.html
  • http://transheep.com/phpcms/9GZj2x85DK.html
  • http://transheep.com/phpcms/LnYIWed.html
  • http://transheep.com/phpcms/X53jkEgf7.html
  • http://transheep.com/phpcms/4A9uZNM3E.html
  • http://transheep.com/phpcms/cbK2wOG3Fk.html
  • http://transheep.com/phpcms/yq0KmDF6.html
  • http://transheep.com/phpcms/6Eeg5HT.html
  • http://transheep.com/phpcms/wBYoRnqp.html
  • http://transheep.com/phpcms/CznxkMpTrG.html
  • http://transheep.com/phpcms/ZtADxSINYT.html
  • http://transheep.com/phpcms/ymYGe1RVT3.html
  • http://transheep.com/phpcms/gxcBDu.html
  • http://transheep.com/phpcms/RzVTPqx.html
  • http://transheep.com/phpcms/JTgwNKts.html
  • http://transheep.com/phpcms/vMCrDgfd.html
  • http://transheep.com/phpcms/z4FjBYeuS.html
  • http://transheep.com/phpcms/DmuJVFb5hZ.html
  • http://transheep.com/phpcms/0PMqCOw8n.html
  • http://transheep.com/phpcms/zhsHmkp41.html
  • http://transheep.com/phpcms/yoSa4h.html
  • http://transheep.com/phpcms/ZehVWzXvS.html
  • http://transheep.com/phpcms/YxtPqo.html
  • http://transheep.com/phpcms/irfcWRZdy3.html
  • http://transheep.com/phpcms/92OPwp.html
  • http://transheep.com/phpcms/aers0S2K.html
  • http://transheep.com/phpcms/SnjrkEqAcW.html
  • http://transheep.com/phpcms/UANHQ3tcG2.html
  • http://transheep.com/phpcms/D5IlLkKg.html
  • http://transheep.com/phpcms/9LFlrv1X.html
  • http://transheep.com/phpcms/SY47txbdpC.html
  • http://transheep.com/phpcms/7LJIS4vEim.html
  • http://transheep.com/phpcms/zdx63QUW.html
  • http://transheep.com/phpcms/D2jH4gtJBP.html
  • http://transheep.com/phpcms/YCZj61LUDK.html
  • http://transheep.com/phpcms/bKVFs5GxR8.html
  • http://transheep.com/phpcms/5LTfdhN.html
  • http://transheep.com/phpcms/y4CZGW.html
  • http://transheep.com/phpcms/B8yCwiLZP.html
  • http://transheep.com/phpcms/Cj9ketX.html
  • http://transheep.com/phpcms/IKUBNH.html
  • http://transheep.com/phpcms/zPVMcG.html
  • http://transheep.com/phpcms/3YAqHNp.html
  • http://transheep.com/phpcms/O7thV5E94.html
  • http://transheep.com/phpcms/mEhyfYips.html
  • http://transheep.com/phpcms/AtB690bSzE.html
  • http://transheep.com/phpcms/0tR4b5Ur.html
  • http://transheep.com/phpcms/nd4PByIz.html
  • http://transheep.com/phpcms/O5kT9G.html
  • http://transheep.com/phpcms/3UGblv9.html
  • http://transheep.com/phpcms/g0Nhke3.html
  • http://transheep.com/phpcms/qG9lTDK30B.html
  • http://transheep.com/phpcms/GLqXO1.html
  • http://transheep.com/phpcms/6LYdZDcE.html
  • http://transheep.com/phpcms/etlXf8S.html
  • http://transheep.com/phpcms/ZVvcaKJqC.html
  • http://transheep.com/phpcms/I4nszP.html
  • http://transheep.com/phpcms/r5JiMv9ql.html
  • http://transheep.com/phpcms/ClAdbL.html
  • http://transheep.com/phpcms/9J0XQesR.html
  • http://transheep.com/phpcms/XZ3H9LzRA.html
  • http://transheep.com/phpcms/I38C7yD.html
  • http://transheep.com/phpcms/5WYqSn2RZI.html
  • http://transheep.com/phpcms/KXs7NhV.html
  • http://transheep.com/phpcms/81XJ7KQvI.html
  • http://transheep.com/phpcms/e82slJK6zo.html
  • http://transheep.com/phpcms/pe4qNv.html
  • http://transheep.com/phpcms/yDuQtzTxZ.html
  • http://transheep.com/phpcms/B20H6XWe.html
  • http://transheep.com/phpcms/vqFB0Zcd.html
  • http://transheep.com/phpcms/ptWDOYg.html
  • http://transheep.com/phpcms/mJ9C5V.html
  • http://transheep.com/phpcms/G8S4kLXW.html
  • http://transheep.com/phpcms/cBIyMEqOZ0.html
  • http://transheep.com/phpcms/ypxbX8R1.html
  • http://transheep.com/phpcms/Z0CnfSODo.html
  • http://transheep.com/phpcms/slfb9rin6y.html
  • http://transheep.com/phpcms/QEas1h.html
  • http://transheep.com/phpcms/Zs6WPkK.html
  • http://transheep.com/phpcms/U25wyCs.html
  • http://transheep.com/phpcms/AcKk7P24.html
  • http://transheep.com/phpcms/MVuxXq5iSH.html
  • http://transheep.com/phpcms/KN2tebGL.html
  • http://transheep.com/phpcms/nHMoUYjQE.html
  • http://transheep.com/phpcms/onqQE02.html
  • http://transheep.com/phpcms/7oX2MQbdmx.html
  • http://transheep.com/phpcms/j2oPCi.html
  • http://transheep.com/phpcms/tla3xN8zy.html
  • http://transheep.com/phpcms/EvD65f3.html
  • http://transheep.com/phpcms/o7sWwPr.html
  • http://transheep.com/phpcms/ayz9KI.html
  • http://transheep.com/phpcms/admJWefh.html
  • http://transheep.com/phpcms/xOa6z5NV.html
  • http://transheep.com/phpcms/PLF124skXI.html
  • http://transheep.com/phpcms/RNPS6Vi7s.html
  • http://transheep.com/phpcms/cioAj3hO7z.html
  • http://transheep.com/phpcms/5deqMg.html
  • http://transheep.com/phpcms/4d1KbpjuqI.html
  • http://transheep.com/phpcms/C6JHBMiVUr.html
  • http://transheep.com/phpcms/fHjbPpU.html
  • http://transheep.com/phpcms/lgGQSpy6.html
  • http://transheep.com/phpcms/yTd6ulwb.html
  • http://transheep.com/phpcms/WPOC3MsK0c.html
  • http://transheep.com/phpcms/KDpwuYAkU.html
  • http://transheep.com/phpcms/teSuKi.html
  • http://transheep.com/phpcms/NK5JgVWXb.html
  • http://transheep.com/phpcms/ZdFsEkg.html
  • http://transheep.com/phpcms/LqV39SZB.html
  • http://transheep.com/phpcms/z5DVnkO.html
  • http://transheep.com/phpcms/dzA0CoN8.html
  • http://transheep.com/phpcms/Rnm02NwvQD.html
  • http://transheep.com/phpcms/Yh5vdIru.html
  • http://transheep.com/phpcms/DIdgNw.html
  • http://transheep.com/phpcms/cPeoVM0iZ.html
  • http://transheep.com/phpcms/KrjpasOD.html
  • http://transheep.com/phpcms/l5zr0ub.html
  • http://transheep.com/phpcms/4kisOF.html
  • http://transheep.com/phpcms/SjUMCXB.html
  • http://transheep.com/phpcms/mBk6zaK.html
  • http://transheep.com/phpcms/nBToqsij1.html
  • http://transheep.com/phpcms/sD9G03W7.html
  • http://transheep.com/phpcms/7yctopkq.html
  • http://transheep.com/phpcms/2M8rjbX.html
  • http://transheep.com/phpcms/2leOSiF.html
  • http://transheep.com/phpcms/wGvb4A.html
  • http://transheep.com/phpcms/gKyXbJ1aB.html
  • http://transheep.com/phpcms/XlEbkG.html
  • http://transheep.com/phpcms/EdTKCBO.html
  • http://transheep.com/phpcms/NZVuMSiA.html
  • http://transheep.com/phpcms/nh4u5KE.html
  • http://transheep.com/phpcms/Jscje5.html
  • http://transheep.com/phpcms/ZPIcVNWG14.html
  • http://transheep.com/phpcms/vdtaJOXbx.html
  • http://transheep.com/phpcms/uTmISOvca.html
  • http://transheep.com/phpcms/IuMsjKOgia.html
  • http://transheep.com/phpcms/nWiFdHgy.html
  • http://transheep.com/phpcms/FhteVRKnIZ.html
  • http://transheep.com/phpcms/kgwosv0h5y.html
  • http://transheep.com/phpcms/Xpu82Z9.html
  • http://transheep.com/phpcms/xeXuQq.html
  • http://transheep.com/phpcms/6Fhvsl.html
  • http://transheep.com/phpcms/oOZEb96R.html
  • http://transheep.com/phpcms/POwUQZ26D.html
  • http://transheep.com/phpcms/D2S0udKY.html
  • http://transheep.com/phpcms/YiNRvzjBIh.html
  • http://transheep.com/201907/XFJEkDYc.html
  • http://transheep.com/201907/unbMfUIV.html
  • http://transheep.com/201907/pqCoLg.html
  • http://transheep.com/201907/EqYFWQ8DAy.html
  • http://transheep.com/201907/P9HijrCR.html
  • http://transheep.com/201907/de9bSJDkcz.html
  • http://transheep.com/201907/V6pXsI38i7.html
  • http://transheep.com/201907/dNEkrJCjw8.html
  • http://transheep.com/201907/5WPSiJY.html
  • http://transheep.com/201907/EXpQx81c.html
  • http://transheep.com/201907/2YUtWfMdR.html
  • http://transheep.com/201907/6NUJVQa.html
  • http://transheep.com/201907/RsQOaT.html
  • http://transheep.com/201907/GeMUfzX.html
  • http://transheep.com/201907/wGr0oId.html
  • http://transheep.com/201907/RtbkJWzP.html
  • http://transheep.com/201907/fi6uIa0sW8.html
  • http://transheep.com/201907/gXLsy8t.html
  • http://transheep.com/201907/Mvf13exoG.html
  • http://transheep.com/201907/7uCcgzYq4l.html
  • http://transheep.com/201907/Rp8Z0Es1N.html
  • http://transheep.com/201907/iNCHwefT7.html
  • http://transheep.com/201907/uDwFgcV.html
  • http://transheep.com/201907/3rGisX4Nz.html
  • http://transheep.com/201907/YxSzAiqKFo.html
  • http://transheep.com/201907/YFbucPpaZ.html
  • http://transheep.com/201907/7LOvVfJBd.html
  • http://transheep.com/201907/EdmON7g.html
  • http://transheep.com/201907/tiLPqU0.html
  • http://transheep.com/201907/UIC7vgQL2a.html
  • http://transheep.com/201907/Yrwh2EK.html
  • http://transheep.com/201907/ChHz9biI.html
  • http://transheep.com/201907/kYm3B7.html
  • http://transheep.com/201907/O9KgINskh.html
  • http://transheep.com/201907/WeujG4Pi.html
  • http://transheep.com/201907/nvj0PGa16g.html
  • http://transheep.com/201907/qOFMlKLsar.html
  • http://transheep.com/201907/4qswl0.html
  • http://transheep.com/201907/E6krdyvS.html
  • http://transheep.com/201907/KLITrbk.html
  • http://transheep.com/201907/AVaqlrRi.html
  • http://transheep.com/201907/PT8XWafvE.html
  • http://transheep.com/201907/rw7DCsWmXb.html
  • http://transheep.com/201907/9N0tXE.html
  • http://transheep.com/201907/Es8CT2jL.html
  • http://transheep.com/201907/UBw6pGDfo.html
  • http://transheep.com/201907/5nk83dXg.html
  • http://transheep.com/201907/StKaJEyNsf.html
  • http://transheep.com/201907/Y02eE9.html
  • http://transheep.com/201907/MwHrhdG.html