• 哈爾濱醫(yī)科大學(xué)附屬第二臨床醫(yī)院普外二科(哈爾濱150086);

目的 　探討將真核表達載體pcDNA3.1-VEGF-C重組質(zhì)粒轉(zhuǎn)染入人乳腺癌MCF-7細(xì)胞后VEGF-C表達的變化。

方法 　通過脂質(zhì)體介導(dǎo)方法，將構(gòu)建好含有VEGF-C的重組質(zhì)粒轉(zhuǎn)染入人乳腺癌MCF-7細(xì)胞中，新霉素（G418）篩選得到穩(wěn)定轉(zhuǎn)染細(xì)胞系，RT-PCR和Western blot方法檢測穩(wěn)定轉(zhuǎn)染后細(xì)胞中VEGF-C mRNA和蛋白的表達。

結(jié)果 　成功轉(zhuǎn)染并獲得穩(wěn)定高表達VEGF-C的乳腺癌MCF-7細(xì)胞系， 其轉(zhuǎn)染組VEGF-C mRNA相對吸光度值（12.382±2.183）較空載組(6.039±1.950)顯著上調(diào)(P＜0.01)。Western blot檢測轉(zhuǎn)染組VEGF-C蛋白相對灰度值(0.971±0.186)較空載組(0.594±0.196)明顯上調(diào)(P＜0.05)。 結(jié)論 　脂質(zhì)體介導(dǎo)重組質(zhì)粒pcDNA3.1-VEGF-C轉(zhuǎn)染入人乳腺癌MCF-7細(xì)胞中可顯著增加VEGF-C表達水平。

引用本文： 單明,張建國,郭寶良. VEGF-C基因轉(zhuǎn)染對人乳腺癌MCF-7細(xì)胞VEGF-C表達的影響. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(10): 739-742. doi: 復(fù)制

1. 　Noguchi T, Takeno S, Shibata T, et al. VEGF-C expression correlates with histological differentiation and metastasis in squamous cell carcinoma of the esophagus [Ｊ]. Oncol Rep, 2002; 9(5)∶995.
2. 　Sambrook J, Russell DW, 黃培堂, 等譯. 分子克隆實驗指南 [M]. 第3版. 北京: 科學(xué)出版社, 2002∶1713~1726.
3. 　Ishikawa M, Kitayama J, Kazama S, et al. Expression of vascular endothelial growth factor C and D (VEGF-C and -D) is an important risk factor for lymphatic metastasis in undifferentiated early gastric carcinoma [Ｊ]. Jpn J Clin Oncol, 2003; 33(1)∶21.
4. 　王凱峰， 葉勝龍， 宋麗杰, 等. 不同轉(zhuǎn)移潛能的小鼠肝癌淋巴道轉(zhuǎn)移和淋巴管生成的研究 [Ｊ]. 中華肝臟病雜志, 2006; 14(3)∶187.
5. 　He Y, Rajantie I, Pajusola K, et al. Vascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels [Ｊ]. Cancer Res, 2005; 65(11)∶4739.
6. 　Karkkainen MJ, Haiko P, Sainio K, et al. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins [Ｊ]. Nat Immunol, 2004; 5(1)∶74.
7. 　Auvinen MI, Sihvo EI, Ruohtula T, et al. Incipient angiogenesis in Barrett’s epithelium and lymphangiogenesis in Barrett’s adenocarcinoma [Ｊ]. J Clin Oncol, 2002; 20(13)∶2971.
8. 　Bachelder RE, Crago A, Chung J, et al. Vascular endothelial growth factor is an autocrine survival factor for neuropilin-expressing breast carcinoma cells [Ｊ]. Cancer Res, 2001; 61(15)∶5736.
9. 　Onogawa S, Kitadai Y, Tanaka S, et al. Regulation of vascular endothelial growth factor (VEGF)-C and VEGF-D expression by the organ microenvironment in human colon carcinoma [Ｊ]. Eur J Cancer, 2004; 40(10)∶1604.
10. 　Brusselmans K, Bono F, Collen D, et al. A novel role for vascular endothelial growth factor as an autocrine survival factor for embryonic stem cells during hypoxia [Ｊ]. J Biol Chem, 2005; 280(5)∶3493.
11. 　Mkinen T, Veikkola T, Mustjoki S, et al. Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3 [Ｊ]. EMBO J, 2001; 20(17)∶4762.
12. 　Oliver G, Detmar M. The rediscovery of the lymphatic system: old and new insights into the development and biological function of the lymphatic vasculature [Ｊ]. Genes Dev, 2002; 16(7)∶773.
13. 　Dixelius J, Makinen T, Wirzenius M, et al. Ligand-induced vascular endothelial growth factor receptor-3 (VEGFR-3) heterodimerization with VEGFR-2 in primary lymphatic endothelial cells regulates tyrosine phosphorylation sites [Ｊ]. J Biol Chem, 2003; 278(42)∶40973.
14. 　梁燕, 姜軍, 王清良, 等. 自制組織芯片研究VEGF-C在乳腺癌中的表達及其意義 [Ｊ].中國普外基礎(chǔ)與臨床雜志, 2006; 13(5)∶580.
15. 　Shibuya M, Claesson-Welsh L. Singnal transduction by VEGF receptors in regulation of angiogenesis and lymphangiogenesis [Ｊ]. Exp Cell Res, 2006; 312(5)∶549.
16. 　Paavonen K, Horelli-Kuitunen N, Chilov D, et al. Novel human vascular endothelial growth factor genes VEGF-B and VEGF-C localize to chromosomes 11q13 and 4q34, respectively [Ｊ]. Circulation, 1996; 93(6)∶1079.
17. 　潘劍, 華成舸, 溫玉明, 等. 人血管內(nèi)皮生長因子-C基因編碼區(qū)cDNA的克隆與測序 [Ｊ]. 華西口腔醫(yī)學(xué)雜志, 2003; 21(4)∶318.
18. 　Joukov V, Sorsa T, Kumar V, et al. Proteolytic processing regulates receptor specificity and activity of VEGF-C [Ｊ]. EMBO J, 1997; 16(3)∶3898.
19. 　Ropert C. Liposomes as a gene delivery system [Ｊ]. Braz J Med Biol Res, 1999; 32(2)∶163.
20. 　張克君, 高焱明, 楊金鏞, 等. IκB-α基因轉(zhuǎn)染HCC9204細(xì)胞對NF-κB和MMP-9表達的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2007; 14(2)∶185.
  1. 1. 　Noguchi T, Takeno S, Shibata T, et al. VEGF-C expression correlates with histological differentiation and metastasis in squamous cell carcinoma of the esophagus [Ｊ]. Oncol Rep, 2002; 9(5)∶995.
  2. 2. 　Sambrook J, Russell DW, 黃培堂, 等譯. 分子克隆實驗指南 [M]. 第3版. 北京: 科學(xué)出版社, 2002∶1713~1726.
  3. 3. 　Ishikawa M, Kitayama J, Kazama S, et al. Expression of vascular endothelial growth factor C and D (VEGF-C and -D) is an important risk factor for lymphatic metastasis in undifferentiated early gastric carcinoma [Ｊ]. Jpn J Clin Oncol, 2003; 33(1)∶21.
  4. 4. 　王凱峰， 葉勝龍， 宋麗杰, 等. 不同轉(zhuǎn)移潛能的小鼠肝癌淋巴道轉(zhuǎn)移和淋巴管生成的研究 [Ｊ]. 中華肝臟病雜志, 2006; 14(3)∶187.
  5. 5. 　He Y, Rajantie I, Pajusola K, et al. Vascular endothelial cell growth factor receptor 3-mediated activation of lymphatic endothelium is crucial for tumor cell entry and spread via lymphatic vessels [Ｊ]. Cancer Res, 2005; 65(11)∶4739.
  6. 6. 　Karkkainen MJ, Haiko P, Sainio K, et al. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins [Ｊ]. Nat Immunol, 2004; 5(1)∶74.
  7. 7. 　Auvinen MI, Sihvo EI, Ruohtula T, et al. Incipient angiogenesis in Barrett’s epithelium and lymphangiogenesis in Barrett’s adenocarcinoma [Ｊ]. J Clin Oncol, 2002; 20(13)∶2971.
  8. 8. 　Bachelder RE, Crago A, Chung J, et al. Vascular endothelial growth factor is an autocrine survival factor for neuropilin-expressing breast carcinoma cells [Ｊ]. Cancer Res, 2001; 61(15)∶5736.
  9. 9. 　Onogawa S, Kitadai Y, Tanaka S, et al. Regulation of vascular endothelial growth factor (VEGF)-C and VEGF-D expression by the organ microenvironment in human colon carcinoma [Ｊ]. Eur J Cancer, 2004; 40(10)∶1604.
  10. 10. 　Brusselmans K, Bono F, Collen D, et al. A novel role for vascular endothelial growth factor as an autocrine survival factor for embryonic stem cells during hypoxia [Ｊ]. J Biol Chem, 2005; 280(5)∶3493.
  11. 11. 　Mkinen T, Veikkola T, Mustjoki S, et al. Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3 [Ｊ]. EMBO J, 2001; 20(17)∶4762.
  12. 12. 　Oliver G, Detmar M. The rediscovery of the lymphatic system: old and new insights into the development and biological function of the lymphatic vasculature [Ｊ]. Genes Dev, 2002; 16(7)∶773.
  13. 13. 　Dixelius J, Makinen T, Wirzenius M, et al. Ligand-induced vascular endothelial growth factor receptor-3 (VEGFR-3) heterodimerization with VEGFR-2 in primary lymphatic endothelial cells regulates tyrosine phosphorylation sites [Ｊ]. J Biol Chem, 2003; 278(42)∶40973.
  14. 14. 　梁燕, 姜軍, 王清良, 等. 自制組織芯片研究VEGF-C在乳腺癌中的表達及其意義 [Ｊ].中國普外基礎(chǔ)與臨床雜志, 2006; 13(5)∶580.
  15. 15. 　Shibuya M, Claesson-Welsh L. Singnal transduction by VEGF receptors in regulation of angiogenesis and lymphangiogenesis [Ｊ]. Exp Cell Res, 2006; 312(5)∶549.
  16. 16. 　Paavonen K, Horelli-Kuitunen N, Chilov D, et al. Novel human vascular endothelial growth factor genes VEGF-B and VEGF-C localize to chromosomes 11q13 and 4q34, respectively [Ｊ]. Circulation, 1996; 93(6)∶1079.
  17. 17. 　潘劍, 華成舸, 溫玉明, 等. 人血管內(nèi)皮生長因子-C基因編碼區(qū)cDNA的克隆與測序 [Ｊ]. 華西口腔醫(yī)學(xué)雜志, 2003; 21(4)∶318.
  18. 18. 　Joukov V, Sorsa T, Kumar V, et al. Proteolytic processing regulates receptor specificity and activity of VEGF-C [Ｊ]. EMBO J, 1997; 16(3)∶3898.
  19. 19. 　Ropert C. Liposomes as a gene delivery system [Ｊ]. Braz J Med Biol Res, 1999; 32(2)∶163.
  20. 20. 　張克君, 高焱明, 楊金鏞, 等. IκB-α基因轉(zhuǎn)染HCC9204細(xì)胞對NF-κB和MMP-9表達的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2007; 14(2)∶185.