Inhibitory effect of RNA interference targeting vascular epithelial growth factor-C on proliferation of human colon cancer cell line LOVO in vitro and In vivo
10.3760/cma.j.issn.1008-1372.2010.08.001
- VernacularTitle:shRNA沉默VEGF-C基因对大肠癌淋巴管生成和淋巴结转移的影响
- Author:
Xiaowen HE
;
Ting LOI
;
Daojin CHEN
- Publication Type:Journal Article
- Keywords:
RNA,small interfering;
Gene silencing;
Vascular endothelial growth factor C/GE;
Intestinal neoplasms/ME/PA;
Lymphatic metastasis;
Lymphangiogenesis
- From:
Journal of Chinese Physician
2010;12(8):1009-1013
- CountryChina
- Language:Chinese
-
Abstract:
Objective This study was to explore the inhibitory effect of shRNA-VEGF - C on growth of human colon cancer cell line Lovo in vitro and vivo. Methods Recombinant VEGF-C short hairpin RNA (shRNA) plasmid was constructed and transfected into Lovo cells. The expression of VEGF-C was detected at mRNA and protein levels by real-time reverse transcription-polymerase chain reaction (RT-PCR). In vivo study, xenograft tumors were established by injecting LOVO cells into nude mice, then shR-NA-VEGF-C were injected into the tumors, the tumor volume and weight and the incidences of lymph node metastasis were detected. Immunohistochemical staining was used to detect the lymphatic microvessel density of colon cancer tissues. Results After transfection of shRNA-VEGF-C, the mRNA of VEGF-C in Lovo cells were down-regulated. Four weeks after injection, the tumor volume and tumor weight in VEGF-C-shR-NA group were significantly smaller than that in empty plasmid group and NS group [(324. 9 ± 64. 8 ) mm3 vs. (553.5±90. 1)mm3 and (570. 1±85.4)mm3; (3.01 ±0.55)g vs (4.65 ±0.65)g and (4.75 ±0. 75)g]. The incidences of lymph node metastasis (30. 1% ) were significantly inhibited compared with empty plasmid group (50. 2% ) and normal saline group (53. 1% ). In shRNA-VEGF-C group, and microlymphatic density (15.5 ± 6. 90) was also decreased compared with empty plasmid group (24. 18 ±6. 45 ), and normal saline group (29. 59 ± 8. 21 ) ( all P <0. 01 ). Conclusion shRNA-VEGF-C can inhibit the growth of LOVO cells in vitro and vivo. VEGF-C may inhibit the lymph node metastasis of colon cancer by suppressing lymphangiogenesis.
