Effect of VEGF-C Gene Transfection on Expression of VEGF-C in Human Breast Cancer MCF-7 Cell_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

SHAN Ming , ZHANG Jianguo , GUO Baoliang

The Second Department of General Surgery， The Second Affiliated Clinical Hospital of Harbin Medical University， Harbin 150086， ChinaCorresponding Author： ZHANG Jian-guo， E-mail： zhangjiang43@hotmail.com;

Corresponding author：

Keywords：

MCF-7 cell line; Eukaryotic expression; Plasmid; Vascular endothelial growth factor-C; Gene transfection

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Objective 　To explore the effect of vascular endothelial growth factor-C (VEGF-C) gene transfection on the expression level of VEGF-C in human breast cancer MCF-7 cell.
Methods 　The constructed VEGF-C gene eukaryotic expression vector was transfected into the human breast cancer MCF-7 cell by using lipofectamine transfection reagents， and the positive cell clones were obtained through G418 selection after transfection. The expressions of VEGF-C mRNA and protein were detected by RT-PCR and Western blot respectively.
Results 　Following the transfection of the VEGF-C recombination plasmid, there were significant differences on the expression levels of VEGF-C mRNA and protein between pcDNA3.1-VEGF-C transfection group and pcDNA3.1 transfection group （12.382±2.183 vs 6.039±1.950, P＜0.01; 0.971±0.186 vs 0.594±0.196, P＜0.05).
Conclusion 　With the transfection of pcDNA3.1-VEGF-C vector by using the liposome, the expression levels of VEGF-C mRNA and protein rise up in breast cancer MCF-7 cell.

Citation： SHAN Ming,ZHANG Jianguo,GUO Baoliang. Effect of VEGF-C Gene Transfection on Expression of VEGF-C in Human Breast Cancer MCF-7 Cell. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2008, 15(10): 739-742. doi: Copy

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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.
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14.	梁燕, 姜军, 王清良, 等. 自制组织芯片研究VEGF-C在乳腺癌中的表达及其意义 [Ｊ].中国普外基础与临床杂志, 2006; 13(5)∶580.
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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.	潘剑, 华成舸, 温玉明, 等. 人血管内皮生长因子-C基因编码区cDNA的克隆与测序 [Ｊ]. 华西口腔医学杂志, 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.	张克君, 高焱明, 杨金镛, 等. IκB-α基因转染HCC9204细胞对NF-κB和MMP-9表达的影响 [Ｊ]. 中国普外基础与临床杂志, 2007; 14(2)∶185.

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, 黄培堂, 等译. 分子克隆实验指南 [M]. 第3版. 北京: 科学出版社, 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. 　王凯峰，叶胜龙，宋丽杰, 等. 不同转移潜能的小鼠肝癌淋巴道转移和淋巴管生成的研究 [Ｊ]. 中华肝脏病杂志, 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, 姜军, 王清良, 等. 自制组织芯片研究VEGF-C在乳腺癌中的表达及其意义 [Ｊ].中国普外基础与临床杂志, 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. 　潘剑, 华成舸, 温玉明, 等. 人血管内皮生长因子-C基因编码区cDNA的克隆与测序 [Ｊ]. 华西口腔医学杂志, 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. 　张克君, 高焱明, 杨金镛, 等. IκB-α基因转染HCC9204细胞对NF-κB和MMP-9表达的影响 [Ｊ]. 中国普外基础与临床杂志, 2007; 14(2)∶185.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Effect of VEGF-C Gene Transfection on Expression of VEGF-C in Human Breast Cancer MCF-7 Cell

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