长链非编码RNA-MALAT1对结直肠癌细胞介导的血管形成的影响

Effect of lncRNA-MALAT1 on Colorectal Cancer Cells Mediated Angiogenesis

目的观察长链非编码RNA-肺腺癌转移相关转录因子1(MALAT1)对结直肠癌细胞介导的血管形成的影响,并初步探索其中可能的机理。方法在结直肠癌细胞株SW48中通过转染质粒过表达MALAT1经常规培养或乏氧培养后,收集培养液上清利用酶联免疫吸附试验(ELISA)检测其中血管内皮生长因子(VEGF)的含量,并利用此培养液上清孵育脐静脉内皮细胞(HUVEC),检测其对HUVEC形成血管微管能力的影响;同时收集SW48细胞通过蛋白质免疫印迹(Western blot)法检测其中缺氧诱导因子-1α(HIF-1α)的表达。结果在SW48中过表达MALAT1后,常规培养条件下其培养液上清中的VEGF含量为(514±32)mg/L,较对照组的(110±14)mg/L明显增高(P<0.05),乏氧培养条件下MALAT1组VEGF含量为(928±18)mg/L,较对照组的(230±21)mg/L也有明显增高(P<0.05);利用上述培养液上清孵育HUVEC,过表达MALAT1可促进其体外的成管能力;同时Western blot法证实过表达MALAT1可促进HIF-1α蛋白的表达。结论过表达MALAT1可促进结直肠癌细胞介导的血管形成,其可能作为结直肠癌新的药物治疗靶点。

Objective To observe the effect of lncRNA-metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） on colorectal cancer cells-induced angiogenesis, and explore the potential underlying mechanism. Methods MALAT1 was overexpressed in colorectal cancer cells SW48 by plasmids transfection, then SW48 cells were cultured at normoxia or hypoxia conditions. The culture media was collected, and the concentration of vascular endothelial growth factor （VEGF） in the media was measured by the enzyme-linked immuno sorbent assay （ELISA）, and the human umbilical vein endothelial cells （HUVEC） were incubated with the media collected above. Meanwhile, the expression ofhypoxia-inducible factor-1α （HIF-1α） in SW48 cells was detected by western blot. Results Overexpression of MALAT1 increased the VEGF level in the culture media, normoxia： the MALAT1 group （514±32） mg/L vs. the control group （110± 14） mg/L, P〈0.05; hypoxia： the MALAT 1 group （928±18） mg/L vs. the control group （230±21） rag/L, P〈0.05. Meanwhile, the tube formation activity of HUVEC was enhanced, and the expression of HIF- 1α was elevated in the MALAT1 group by western blot. Conclusion Overexpression of MALAT1 could promote colorectal cancer cells- mediated angiogenesis, it may be developed as a new drug target for colorectal cancer treatment.