目的 研究組織因子途徑抑制物-2(TFPI-2)基因?qū)θ艘认侔┘?xì)胞系Panc-1細(xì)胞侵襲能力的影響.方法 通過(guò)脂質(zhì)體法將TFPI-2基因轉(zhuǎn)染到Panc-1細(xì)胞, 用RT-PCR和Western blot分別檢測(cè)轉(zhuǎn)染前、后TFPI-2 mRNA和相應(yīng)蛋白的表達(dá).Boyden小室法測(cè)定Panc-1-TFPI-2、Panc-1-V和Panc-1-P 3組細(xì)胞的體外侵襲能力的變化,同時(shí)將3組細(xì)胞分別接種裸鼠,觀察其體內(nèi)腫瘤生長(zhǎng)及轉(zhuǎn)移情況.結(jié)果 轉(zhuǎn)染成功的Panc-1細(xì)胞有TFPI-2 mRNA及相應(yīng)蛋白的表達(dá); Panc-1-TFPI-2組、Panc-1-V組和Panc-1-P組細(xì)胞穿膜細(xì)胞數(shù)分別為(24.4±3.5)個(gè)、(61.3±4.1)個(gè)和(60.2±3.9)個(gè),前者明顯少于后兩者(P＜0.05),提示Panc-1-TFPI-2組細(xì)胞的體外侵襲能力下降.3組細(xì)胞分別接種裸鼠,Panc-1-TFPI-2組腫瘤體積為(438.0±69.8) mm3,與Panc-1-V組的(852.0±102.9) mm3和Panc-1-P組的(831.0±78.1) mm3相比明顯縮小,差異有統(tǒng)計(jì)學(xué)意義(P＜0.05); 鏡下見(jiàn)Panc-1-V組和Panc-1-P組腫瘤組織浸潤(rùn)到肌層,有肝、肺轉(zhuǎn)移灶,而Panc-1-TFPI-2組未見(jiàn)肌層浸潤(rùn)及肝、肺轉(zhuǎn)移.結(jié)論 TFPI-2基因表達(dá)可抑制胰腺癌細(xì)胞系Panc-1的侵襲轉(zhuǎn)移能力,為胰腺癌的基因治療提供了實(shí)驗(yàn)依據(jù).
引用本文： 湯志剛,孫振陽(yáng),胡何節(jié),許戈良,陳炯,李建生. TFPI-2 對(duì)胰腺癌細(xì)胞體內(nèi)和體外侵襲能力的影響. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 15(1): 46-50. doi: 復(fù)制
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- 1. Wong CM,Ng YL,Lee JM,et al.Tissue factor pathway inhibitor-2 as a frequently silenced tumor suppressor gene in hepatocellular carcinoma[J].Hepatology,2007; 45(5)∶1129.
- 2. Nobeyama Y,Okochi-Takada E,Furuta J,et al.Silencing of tissue factor pathway inhibitor-2 gene in malignant melanomas[J].Int J Cancer,2007; 121(2)∶301.
- 3. Torres-Collado AX,Kisiel W,Iruela-Arispe ML,et al.ADAMTS1 interacts with,cleaves,and modifies the extracellular location of the matrix inhibitor tissue factor pathway inhibitor-2[J].J Biol Chem,2006; 281(26)∶17827.
- 4. Hirata M,Sato T,Tsumagari M,et al.Discoordinate regulation of expression of matrix metalloproteinases and tissue inhibitor of metalloproteinases-3 in bovine endometrial stromal cells on type-Ⅰ collagen gel[J].Biol Pharm Bull,2003; 26(7)∶1013.
- 5. Siegling S,Thyzel E,Brinkmann T,et al.Genetic variations in the human TFPI-2 gene and its promoter region by denaturing high performance liquid chromatography[J].Thromb Res,2004; 113(5)∶341.
- 6. Chen XY,Li JS,Ma J,et al.Potential role of novel hepatocellular carcinoma-associated gene IDD 01 in promoting tumorigenesis of HepG2 cell line[J].Chin Med J (Engl),2006; 119(20)∶1709.
- 7. Tanaka Y,Utsumi J,Matsui M,et al.Purification,molecular cloning,and expression of a novel growth-promoting factor for retinal pigment epithelial cells,REF-1/TFPI-2[J].Invest Ophthalmol Vis Sci,2004; 45(1)∶245.
- 8. Jiang P,Watanabe H,Okada G,et al.Diagnostic utility of aberrant methylation of tissue factor pathway inhibitor 2 in pure pancreatic juice for pancreatic carcinoma[J].Cancer Sci,2006; 97(11)∶1267.
- 9. Neaud V,Hisaka T,Monvoisin A,et al.Paradoxical pro-invasive effect of the serine proteinase inhibitor tissue factor pathway inhibitor-2 on human hepatocellular carcinoma cells[J].J Biol Chem,2000; 275(45)∶35565.
- 10. Blindt R,Vogt F,Lamby D,et al.Characterization of differential gene expression in quiescent and invasive human arterial smooth muscle cells[J].J Vasc Res,2002; 39(4)∶340.
- 11. Jin M,Udagawa K,Miyagi E,et al.Expression of serine proteinase inhibitor PP5/TFPI-2/MSPI decreases the invasive potential of human choriocarcinoma cells in vitro and in vivo[J].Gynecol Oncol,2001; 83(2)∶325.
- 12. Kong D,Ma D,Bai H,et al.Expression and characterization of the first kunitz domain of human tissue factor pathway inhibitor-2[J].Biochem Biophys Res Commun,2004; 324(4)∶1179.
- 13. Konduri SD,Tasiou A,Chandrasekar N,et al.Role of tissue factor pathway inhibitor-2 (TFPI-2) in amelanotic melanoma (C-32) invasion[J].Clin Exp Metastasis,2000; 18(4)∶303.
- 14. Ivanciu L,Gerard RD,Tang H,et al.Adenovirus-mediated expression of tissue factor pathway inhibitor-2 inhibits endothelial cell migration and angiogenesis[J].Arterioscler Thromb Vasc Biol,2007; 27(2)∶310.
- 15. Iino M,Foster DC,Kisiel W.Quantification and characterization of human endothelial cell-derived tissue factor pathway inhibitor-2[J].Arterioscler Thromb Vasc Biol,1998; 18(1)∶40.
- 16. Rao CN,Reddy P,Reeder DJ,et al.Prokaryotic expression,purification,and reconstitution of biological activities (Antiprotease,antitumor,and heparin-binding) for tissue factor pathway inhibitor-2[J].Biochem Biophys Res Commun,2000; 276(3)∶1286.
- 17. Yanamandra N,Kondraganti S,Gondi CS,et al.Recombinant adeno-associated virus (rAAV) expressing TFPI-2 inhibits invasion,angiogenesis and tumor growth in a human glioblastoma cell line[J].Int J Cancer,2005; 115(6)∶998.
- 18. Chand HS,Du X,Ma D,et al.The effect of human tissue factor pathway inhibitor-2 on the growth and metastasis of fibrosarcoma tumors in athymic mice[J].Blood,2004; 103(3)∶1069.
- 19. Kondraganti S,Gondi CS,Gujrati M,et al.Restoration of tissue factor pathway inhibitor inhibits invasion and tumor growth in vitro and in vivo in a malignant meningioma cell line[J].Int J Oncol,2006; 29(1)∶25.
- 20. Sun Y,Xie M,Liu M,et al.Growth suppression of human laryngeal squamous cell carcinoma by adenovirus-mediated tissue factor pathway inhibitor gene 2[J].Laryngoscope,2006; 116(4)∶596.