RNAi-mediated inhibition of Lgr5 leads to decreased angiogenesis in gastric cancer
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Hong-Qing Xi1,*, Ke-Cheng Zhang1,*, Ji-Yang Li1,*, Jian-Xin Cui1, Yun-He Gao1, Bo Wei1, Dongsheng Huang2 and Lin Chen1
1Department of General Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, China
2Department of General Surgery, Zhejiang Provincial People’s Hospital, Hangzhou 310014, China
*These authors have contributed equally to this work
Lin Chen, email: firstname.lastname@example.org
Dongsheng Huang, email: email@example.com
Keywords: gastric cancer, Lgr5, angiogenesis, RNA interference
Received: November 21, 2016 Accepted: January 11, 2017 Published: February 28, 2017
Leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) is a novel gastric cancer marker. However, it is unclear whether it can play roles in tumor angiogenesis. In this study, we aim to investigate the role of Lgr5 on gastric cancer angiogenesis. Lgr5, VEGF expression levels and microvessel density (MVD) were detected in tumor tissue. Then, Lgr5 mRNA was downregulated by small interference RNA technique. Western blotting and real-time quantitative PCR (qRT-PCR) were performed to detect the expression of Lgr5 and VEGF protein and mRNA in Lgr5 siRNA-transfected gastric cancer cells. The effect of silencing Lgr5 on angiogenesis was examined by assessing human umbilical vein endothelia cell (HUVEC) capillary tube formation. The results indicated that Lgr5 expression was upregulated in gastric cancer and positively correlated with VEGF (r=0.305, P=0.001) and MVD (r=0.312, P=0.001). Silencing of Lgr5 expression resulted in suppression of VEGF mRNA and protein (all P=0.001). Moreover, when HUVECs were stimulated with conditioned medium from Lgr5 siRNA-transfected gastric cancer cells, tube formation was significantly decreased (2.51 ± 0.19 mm/mm2) compared with the treatment with regular cell culture medium (DMEM) (7.34 ± 0.30 mm/mm2) or medium from control siRNA-transfected cells (7.18 ± 0.33 mm/mm2) (all P=0.001). In conclusion, Lgr5 plays important roles in angiogenesis. Lgr5-specific siRNA could be designed into an effective therapeutic agent to inhibit gastric cancer angiogenesis.
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