目的 構(gòu)建穩(wěn)定轉(zhuǎn)染人組織因子(TF)的人胃癌細(xì)胞株SGC7901,研究TF對(duì)胃癌細(xì)胞體外侵襲轉(zhuǎn)移特性的影響。
方法 采用巢式PCR技術(shù)自人胎盤組織克隆目的基因TF,應(yīng)用脂質(zhì)體介導(dǎo)的轉(zhuǎn)染技術(shù)將其導(dǎo)入人胃癌細(xì)胞株SGC7901,采用G418篩選穩(wěn)定表達(dá)TF的細(xì)胞,采用RT-PCR及Western blot法檢測TF mRNA及蛋白的表達(dá),并進(jìn)行Transwell實(shí)驗(yàn)及劃痕實(shí)驗(yàn)觀察細(xì)胞體外侵襲轉(zhuǎn)移能力的變化。
結(jié)果 經(jīng)基因測序證實(shí)成功構(gòu)建了穩(wěn)定、高效表達(dá)TF的人胃癌細(xì)胞系SGC7901/TF。轉(zhuǎn)染組TF mRNA及TF蛋白表達(dá)增高;與陰性對(duì)照組及空白對(duì)照組相比,轉(zhuǎn)染組細(xì)胞的體外侵襲轉(zhuǎn)移能力增強(qiáng)。
結(jié)論 TF能夠增強(qiáng)人胃癌細(xì)胞的體外侵襲轉(zhuǎn)移能力。
引用本文: 俞永江,馬海濤,曹農(nóng),焦作義,李玉民. 組織因子對(duì)人胃癌細(xì)胞體外侵襲轉(zhuǎn)移特性的影響△. 中國普外基礎(chǔ)與臨床雜志, 2012, 19(9): 942-946. doi: 復(fù)制
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2. | Belting M, Ahamed J, Ruf W. Signaling of the tissue factor coagulation pathway in angiogenesis and cancer[J]. Arterioscler Thromb Vasc Biol, 2005, 25(8):1545-1550. |
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6. | Nitori N, Ino Y, Nakanishi Y, et al. Prognostic significance of tissue factor in pancreatic ductal adenocarcinoma[J]. Clin Cancer Res, 2005, 11(7):2531-2539. |
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12. | 陳寧, 張同琳, 姚宏偉. 組織因子對(duì)肝癌細(xì)胞侵襲能力的影響[J]. 中國生物化學(xué)與分子生物學(xué)報(bào), 2006, 22(11):909-913. |
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15. | Versteeg HH, Spek CA, Peppelenbosch MP, et al. Tissue factor and cancer metastasis:the role of intracellular and extracellular signaling pathways[J]. Mol Med, 2004, 10(1-6):6-11. |
16. | Hjortoe GM, Petersen LC, Albrektsen T, et al. Tissue factor-factorⅦa-specific up-regulation of IL-8 expression in MDA-MB-231 cells is mediated by PAR-2 and results in increased cell migration[J]. Blood, 2004, 103(8):3029-3037. |
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24. | 方峻, 湯浩, 夏凌輝, 等. 組織因子siRNA 表達(dá)載體的構(gòu)建及其抑制人神經(jīng)母細(xì)胞瘤細(xì)胞組織因子表達(dá)的研究[J]. 微循環(huán)學(xué), 2007, 17(3):16-18. |
25. | Shibuya M, Claesson-Welsh L. Signal transduction by VEGFreceptors in regulation of angiogenesis and lymphangiogenesis[J]. Exp Cell Res, 2006, 312(5):549-560. |
26. | 方峻, 魏文寧, 劉仲萍, 等. 人組織因子基因表達(dá)載體的構(gòu)建及其在人卵巢癌細(xì)胞系中的表達(dá)[J]. 中國實(shí)驗(yàn)血液學(xué)雜志, 2003, 11(6):579-582. |
- 1. Mackman N. Role of tissue factor in hemostasis, thrombosis, and vascular development[J]. Arterioscler Thromb Vasc Biol, 2004, 24(6):1015-1022.
- 2. Belting M, Ahamed J, Ruf W. Signaling of the tissue factor coagulation pathway in angiogenesis and cancer[J]. Arterioscler Thromb Vasc Biol, 2005, 25(8):1545-1550.
- 3. Yamashita H, Kitayama J, Ishikawa M, et al. Tissue factor expression is a clinical indicator of lymphatic metastasis and poor prognosis in gastric cancer with intestinal phenotype[J]. J Surg Oncol, 2007, 95(4):324-331.
- 4. 焦作義, 茍長征, 曹農(nóng), 等. 組織因子表達(dá)與胃癌微血管生成的關(guān)系及其臨床意義[J]. 癌癥, 2005, 24(7):880-884.
- 5. Nakasaki T, Wada H, Shigemori C, et al. Expression of tissue factor and vascular endothelial growth factor is associated with angiogenesis in colorectal cancer[J]. Am J Hematol, 2002, 69(4):247-254.
- 6. Nitori N, Ino Y, Nakanishi Y, et al. Prognostic significance of tissue factor in pancreatic ductal adenocarcinoma[J]. Clin Cancer Res, 2005, 11(7):2531-2539.
- 7. Francis JL, Biggerstaff J, Amirkhosravi A. Hemostasis and malignancy[J]. Semin Thromb Hemost, 1998, 24(2):93-109.
- 8. Hamada K, Kuratsu J, Saitoh Y, et al. Expression of tissue factor expression correlates with grade of malignancy in human glioma[J]. Cancer, 1996, 77(9):1877-1883.
- 9. Seto S, Onodera H, Kaido T, et al. Tissue factor expression in human colorectal carcinoma:correlation with hepatic metastasis and impact on prognosis[J]. Cancer, 2000, 88(2):295-301.
- 10. Bromberg ME, Sundaram R, Homer RJ, et al. Role of tissue factor in metastasis:functions of the cytoplasmic and extracellular domains of the molecule[J]. Thromb Haemost, 1999, 82(1):88-92.
- 11. 姚宏偉, 萬遠(yuǎn)廉, 吳濤, 等. 組織因子表達(dá)對(duì)人類大腸癌細(xì)胞體外及體內(nèi)侵襲能力的影響[J]. 中華普通外科雜志, 2005, 20(7):442-444.
- 12. 陳寧, 張同琳, 姚宏偉. 組織因子對(duì)肝癌細(xì)胞侵襲能力的影響[J]. 中國生物化學(xué)與分子生物學(xué)報(bào), 2006, 22(11):909-913.
- 13. Brunhilde FH, Yoshikazu T, Barbara M, et al. Inhibition oftissue factor signaling suppresses tumor growth[J]. Blood, 2008, 111(1):190-199.
- 14. Camerer E, Huang W, Coughlin SR. Tissue factor- and factor X-dependent activation of protease-activated receptor 2 by factor Ⅶa[J]. Proc Natl Acad Sci U S A, 2000, 97(10):5255-5260.
- 15. Versteeg HH, Spek CA, Peppelenbosch MP, et al. Tissue factor and cancer metastasis:the role of intracellular and extracellular signaling pathways[J]. Mol Med, 2004, 10(1-6):6-11.
- 16. Hjortoe GM, Petersen LC, Albrektsen T, et al. Tissue factor-factorⅦa-specific up-regulation of IL-8 expression in MDA-MB-231 cells is mediated by PAR-2 and results in increased cell migration[J]. Blood, 2004, 103(8):3029-3037.
- 17. D’Andrea MR, Derian CK, Santulli RJ, et al. Differentialexpression of protease-activated receptors-1 and -2 in stromal fibroblasts of normal, benign, and malignant human tissues[J]. Am J Pathol, 2001, 158(6):2031-2041.
- 18. Dorsam RT, Gutkind JS. G-protein-coupled receptors andcancer[J]. Nat Rev Cancer, 2007, 7(2):79-94.
- 19. Pendurthi UR, Allen KE, Ezban M, et al. FactorⅦa and thrombin induce the expression of Cyr61 and connective tissue growth factor, extracellular matrix signaling proteins that could act as possible downstream mediators in factorⅦa x tissue factor-induced signal transduction[J]. J Biol Chem, 2000, 275(19):14632-14641.
- 20. Zhao W, Wang X, Qu B, et al. Clinical significance of plasma tissue factor pathway and urokinase-type plasminogen activator system in cancer patients[J]. Chin Med J (Engl), 2002, 115(5):702-704.
- 21. Ott I, Fischer EG, Miyagi Y, et al. A role for tissue factor in cell adhesion and migration mediated by interaction with actin-binding protein 280[J]. J Cell Biol, 1998, 140(5):1241-1253.
- 22. 湯堅(jiān)強(qiáng), 萬遠(yuǎn)廉, 劉玉樹, 等. 組織因子促進(jìn)人類大腸癌細(xì)胞的侵襲及與基質(zhì)金屬蛋白酶類的相關(guān)性研究[J]. 北京大學(xué)學(xué)報(bào)(醫(yī)學(xué)版), 2005, 37(3):265-268.
- 23. 辛華, 鄭雅娟, 韓振國. 組織因子在腫瘤細(xì)胞向血管內(nèi)游走過程中的作用[J]. 中國病理生理雜志, 2004, 20(6):960-963.
- 24. 方峻, 湯浩, 夏凌輝, 等. 組織因子siRNA 表達(dá)載體的構(gòu)建及其抑制人神經(jīng)母細(xì)胞瘤細(xì)胞組織因子表達(dá)的研究[J]. 微循環(huán)學(xué), 2007, 17(3):16-18.
- 25. Shibuya M, Claesson-Welsh L. Signal transduction by VEGFreceptors in regulation of angiogenesis and lymphangiogenesis[J]. Exp Cell Res, 2006, 312(5):549-560.
- 26. 方峻, 魏文寧, 劉仲萍, 等. 人組織因子基因表達(dá)載體的構(gòu)建及其在人卵巢癌細(xì)胞系中的表達(dá)[J]. 中國實(shí)驗(yàn)血液學(xué)雜志, 2003, 11(6):579-582.