目的 建立基于人臍靜脈內(nèi)皮細(xì)胞(HUVEC)、纖維蛋白基質(zhì)及微載體的體外三維新生血管模型,在此模型的基礎(chǔ)上,研究CD105在血管新生過(guò)程中的表達(dá)變化及其作用。
方法 分離、純化并培養(yǎng)HUVEC; 建立三維新生血管模型: 將HUVEC包被于微載體(Cytodex-3)上,然后將該包被有HUVEC的微載體包埋于纖維蛋白凝膠基質(zhì)中,在培養(yǎng)體系中內(nèi)皮細(xì)胞生長(zhǎng)因子的作用下完成HUVEC的出芽、分支、連接成網(wǎng)等過(guò)程。通過(guò)逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)檢測(cè)血管新生不同時(shí)期CD105的表達(dá)情況,并利用反義寡核苷酸技術(shù)干預(yù)CD105的表達(dá),觀察抑制CD105表達(dá)后HUVEC與血管新生過(guò)程的表形變化。
結(jié)果 包埋于三維纖維蛋白基質(zhì)中的HUVEC出現(xiàn)出芽、分支、連接成網(wǎng)等典型的血管新生過(guò)程; 在此過(guò)程中,CD105的表達(dá)在出芽早期及出芽分支期明顯增強(qiáng),而在出芽前及成網(wǎng)后的相對(duì)穩(wěn)定期則呈不表達(dá)或弱表達(dá)狀態(tài)。利用反義寡核苷酸抑制CD105表達(dá)后,HUVEC的生長(zhǎng)及新生血管模型中的出芽、分支過(guò)程均受到明顯抑制。
結(jié)論 CD105在血管新生的不同階段表達(dá)水平不同,在出芽、分支期明顯過(guò)表達(dá); CD105參與了血管新生過(guò)程,抑制CD105的表達(dá)可以有效地抑制血管新生。
引用本文: 姚永忠,潘一明,孫喜太,仇毓東,丁義濤. 血管新生過(guò)程中CD105的表達(dá)變化及其作用. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009, 16(10): 824-829. doi: 復(fù)制
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2. | Suh DY. Understanding angiogenesis and its clinical applications [J]. Ann Clin Lab Sci, 2000; 30(3): 227-238. |
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4. | 駱成玉, 祝學(xué)光. 腫瘤血管生成對(duì)微轉(zhuǎn)移影響的研究進(jìn)展 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10(5): 516-518. |
5. | 張盟輝, 孔憲炳, 王巧玲, 等. 射頻消融聯(lián)合亞砷酸局部治療對(duì)兔肝VX2腫瘤MVD和VEGF表達(dá)的影響 [J].中國(guó)普外基礎(chǔ)與臨床雜志, 2007; 14(1): 19-22. |
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7. | Minhajat R, Mori D, Yamasaki F, et al. Endoglin (CD105) expression in angiogenesis of colon cancer: analysis using tissue microarrays and comparison with other endothelial markers [J]. Virchows Arch, 2006; 448(2): 127-134. |
8. | Charpin C, Dales JP, Garcia S, et al. Tumor neoangiogenesis by CD31 and CD105 expression evaluation in breast carcinoma tissue microarrays [J]. Clin Cancer Res, 2004; 10(17): 5815-5819. |
9. | Yao Y, Pan Y, Chen J, et al. Endoglin (CD105) expression in angiogenesis of primary hepatocellular carcinomas: analysis using tissue microarrays and comparisons with CD34 and VEGF [J]. Ann Clin Lab Sci, 2007; 37(1): 39-48. |
10. | Yao Y, Kubota T, Takeuchi H, et al. Prognostic significance of microvessel density determined by an anti-CD105/endoglin monoclonal antibody in astrocytic tumors: comparison with an anti-CD31 monoclonal antibody [J]. Neuropathology, 2005; 25(3): 201-206. |
11. | Costello B, Li C, Duff S, et al. Perfusion of 99Tcm-labeled CD105 Mab into kidneys from patients with renal carcinoma suggests that CD105 is a promising vascular target [J]. Int J Cancer, 2004; 109(3): 436-441. |
12. | Jaffe EA, Nachman RL, Becker CG, et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria [J]. J Clin Invest, 1973; 52(11): 2745-2756. |
13. | Nehls V, Drenckhahn D. A novel, microcarrier-based in vitro assay for rapid and reliable quantification of three-dimensional cell migration and angiogenesis [J]. Microvasc Res, 1995; 50(3): 311-322. |
14. | Xue L, Greisler HP. Angiogenic effect of fibroblast growth factor-1 and vascular endothelial growth factor and their synergism in a novel in vitro quantitative fibrin-based 3-dimensional angiogenesis system [J]. Surgery, 2002; 132(2): 259-267. |
15. | Duff SE, Li C, Garland JM, et al. CD105 is important for angiogenesis: evidence and potential applications [J]. FASEB J, 2003; 17(9): 984-992. |
16. | Dales JP, Garcia S, Andrac L, et al. Prognostic significance of angiogenesis evaluated by CD105 expression compared to CD31 in 905 breast carcinomas: correlation with long-term patient outcome [J]. Int J Oncol, 2004; 24(5): 1197-1204. |
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22. | 馮立民, 姜希宏, 烏新林, 等. survivin反義寡核苷酸誘導(dǎo)膽囊癌細(xì)胞凋亡的研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13(3): 298-301. |
23. | 車向明, 王曙逢, 樊林, 等. E-cadherin反義寡核苷酸對(duì)腫瘤細(xì)胞浸潤(rùn)能力影響的研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13(2): 191-193. |
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25. | Fonsatti E, Altomonte M, Arslan P, et al. Endoglin (CD105): a target for anti-angiogenetic cancer therapy [J]. Curr Drug Targets, 2003; 4(4): 291-296. |
- 1. Folkman J. What is the evidence that tumors are angiogenesis dependent? [J]. J Natl Cancer Inst, 1990; 82(1): 4-6.
- 2. Suh DY. Understanding angiogenesis and its clinical applications [J]. Ann Clin Lab Sci, 2000; 30(3): 227-238.
- 3. Sharma S, Sharma MC, Sarkar C. Morphology of angiogenesis in human cancer: a conceptual overview, histoprognostic perspective and significance of neoangiogenesis [J]. Histopathology, 2005; 46(5): 481-489.
- 4. 駱成玉, 祝學(xué)光. 腫瘤血管生成對(duì)微轉(zhuǎn)移影響的研究進(jìn)展 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10(5): 516-518.
- 5. 張盟輝, 孔憲炳, 王巧玲, 等. 射頻消融聯(lián)合亞砷酸局部治療對(duì)兔肝VX2腫瘤MVD和VEGF表達(dá)的影響 [J].中國(guó)普外基礎(chǔ)與臨床雜志, 2007; 14(1): 19-22.
- 6. Behrem S, Zarkovic K, Eskinja N, et al. Endoglin is a better marker than CD31 in evaluation of angiogenesis in glioblastoma [J]. Croat Med J, 2005; 46(3): 417-422.
- 7. Minhajat R, Mori D, Yamasaki F, et al. Endoglin (CD105) expression in angiogenesis of colon cancer: analysis using tissue microarrays and comparison with other endothelial markers [J]. Virchows Arch, 2006; 448(2): 127-134.
- 8. Charpin C, Dales JP, Garcia S, et al. Tumor neoangiogenesis by CD31 and CD105 expression evaluation in breast carcinoma tissue microarrays [J]. Clin Cancer Res, 2004; 10(17): 5815-5819.
- 9. Yao Y, Pan Y, Chen J, et al. Endoglin (CD105) expression in angiogenesis of primary hepatocellular carcinomas: analysis using tissue microarrays and comparisons with CD34 and VEGF [J]. Ann Clin Lab Sci, 2007; 37(1): 39-48.
- 10. Yao Y, Kubota T, Takeuchi H, et al. Prognostic significance of microvessel density determined by an anti-CD105/endoglin monoclonal antibody in astrocytic tumors: comparison with an anti-CD31 monoclonal antibody [J]. Neuropathology, 2005; 25(3): 201-206.
- 11. Costello B, Li C, Duff S, et al. Perfusion of 99Tcm-labeled CD105 Mab into kidneys from patients with renal carcinoma suggests that CD105 is a promising vascular target [J]. Int J Cancer, 2004; 109(3): 436-441.
- 12. Jaffe EA, Nachman RL, Becker CG, et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria [J]. J Clin Invest, 1973; 52(11): 2745-2756.
- 13. Nehls V, Drenckhahn D. A novel, microcarrier-based in vitro assay for rapid and reliable quantification of three-dimensional cell migration and angiogenesis [J]. Microvasc Res, 1995; 50(3): 311-322.
- 14. Xue L, Greisler HP. Angiogenic effect of fibroblast growth factor-1 and vascular endothelial growth factor and their synergism in a novel in vitro quantitative fibrin-based 3-dimensional angiogenesis system [J]. Surgery, 2002; 132(2): 259-267.
- 15. Duff SE, Li C, Garland JM, et al. CD105 is important for angiogenesis: evidence and potential applications [J]. FASEB J, 2003; 17(9): 984-992.
- 16. Dales JP, Garcia S, Andrac L, et al. Prognostic significance of angiogenesis evaluated by CD105 expression compared to CD31 in 905 breast carcinomas: correlation with long-term patient outcome [J]. Int J Oncol, 2004; 24(5): 1197-1204.
- 17. Wikstrm P, Lissbrant IF, Stattin P, et al. Endoglin (CD105) is expressed on immature blood vessels and is a marker for survival in prostate cancer [J]. Prostate, 2002; 51(4): 268-275.
- 18. 姚衛(wèi)東, 姜惠峰, 曲桂梅, 等. 肝細(xì)胞癌組織中endoglin (CD105)和VEGF 及p57kip2表達(dá)與預(yù)后的關(guān)系 [J]. 腫瘤防治雜志, 2005; 12(5): 360-364.
- 19. Saad RS, Liu YL, Nathan G, et al. Endoglin (CD105) and vascular endothelial growth factor as prognostic markers in colorectal cancer [J]. Mod Pathol, 2004; 17(2): 197-203.
- 20. Saad RS, Jasnosz KM, Tung MY, et al. Endoglin (CD105) expression in endometrial carcinoma [J]. Int J Gynecol Pathol, 2003; 22(3): 248-253.
- 21. Saad RS, El-Gohary Y, Memari E, et al. Endoglin (CD105) and vascular endothelial growth factor as prognostic markers in esophageal adenocarcinoma [J]. Hum Pathol, 2005; 36(9): 955-961.
- 22. 馮立民, 姜希宏, 烏新林, 等. survivin反義寡核苷酸誘導(dǎo)膽囊癌細(xì)胞凋亡的研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13(3): 298-301.
- 23. 車向明, 王曙逢, 樊林, 等. E-cadherin反義寡核苷酸對(duì)腫瘤細(xì)胞浸潤(rùn)能力影響的研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13(2): 191-193.
- 24. Bartsch M, Weeke-Klimp AH, Meijer DK, et al. Cell-specific targeting of lipid-based carriers for ODN and DNA [J]. J Liposome Res, 2005; 15(1-2): 59-92.
- 25. Fonsatti E, Altomonte M, Arslan P, et al. Endoglin (CD105): a target for anti-angiogenetic cancer therapy [J]. Curr Drug Targets, 2003; 4(4): 291-296.