CAJ | 학술논문

目的研究人参皂甙Rg3(简称Rg3)对人鼻咽低分化鳞癌HNE-1细胞体外血管形成的作用.方法采用不同浓度Rg3作用HNE-1细胞,观察HNE-1细胞在Matrigel胶上形成血管网状结构的特点;采用体外管道形成抑制实验检测不同浓度Rg3对HNE-1细胞管道形成的能力;采用免疫组化SP法及Western blot方法检测不同浓度Rg3作用HNE-1细胞48 h后血管内皮细胞生长因子(VEGF)蛋白的表达情况.结果 HNE-1细胞在Matrigel胶上能形成血管网状样结构,并能与内皮细胞连接,共同构成马赛克样血管网状结构.0、50、100、200 μg/ml Rg3作用HNE-1细胞24 h后,管状结构形成数量分别为(75.50±6.86)个、(55.00±11.92)个、(39.75±7.93)个和(24.50±6.25)个,与Rg3浓度呈负相关(r=-0.928,P＜0.01).0、50、100、200 μg/m1 Rg3作用HNE-1细胞48 h后,免疫组化检测显示,VEGF蛋白表达水平明显减弱,分别为0.19±0.03、0.13±0.02、0.11 ±0.01和0.08±0.01,与rg3浓度呈负相关(r=-0.911,P＜0.01);Western blot法检测显示,VEGF表达的A值逐渐减少,分别为119.49、111.51、86.45和38.29,以200μg/ml浓度组作用最为明显.结论 Rg3能够抑制HNE-1细胞体外血管生成拟态的形成,其机制可能与Rg3下调HNE-1细胞中VEGF蛋白的表达有关.
목적 연구인삼조대Rg3(간칭Rg3)대인비인저분화린암HNE-1세포체외혈관형성적작용.방법 채용불동농도Rg3작용HNE-1세포,관찰HNE-1세포재Matrigel효상형성혈관망상결구적특점;채용체외관도형성억제실험검측불동농도Rg3대HNE-1세포관도형성적능력;채용면역조화SP법급Western blot방법검측불동농도Rg3작용HNE-1세포48 h후혈관내피세포생장인자(VEGF)단백적표체정황.결과 HNE-1세포재Matrigel효상능형성혈관망상양결구,병능여내피세포련접,공동구성마새극양혈관망상결구.0、50、100、200 μg/ml Rg3작용HNE-1세포24 h후,관상결구형성수량분별위(75.50±6.86)개、(55.00±11.92)개、(39.75±7.93)개화(24.50±6.25)개,여Rg3농도정부상관(r=-0.928,P＜0.01).0、50、100、200 μg/m1 Rg3작용HNE-1세포48 h후,면역조화검측현시,VEGF단백표체수평명현감약,분별위0.19±0.03、0.13±0.02、0.11 ±0.01화0.08±0.01,여rg3농도정부상관(r=-0.911,P＜0.01);Western blot법검측현시,VEGF표체적A치축점감소,분별위119.49、111.51、86.45화38.29,이200μg/ml농도조작용최위명현.결론 Rg3능구억제HNE-1세포체외혈관생성의태적형성,기궤제가능여Rg3하조HNE-1세포중VEGF단백적표체유관.
Objective To study the anti-angiogenic effect of ginsenoside Rg3 (Rg3 in abbreviation) in human nasopharyngeal carcinoma HNE-1 cells in vitro. Methods The tube-like structure (TLS) formation of HNE-1 cells, cultured in medium with different concentrations of Rg3, was determined by in vitro anti-angiogenic test based on preliminary experiment observing the TLSs formed by HNE-1 cells on Matrigel and their structural characteristics. The VEGF expression level in HNE-1 cells was determined by immunohistochemistry (IHC) and Western-blot test after 48-hour cultured in medium with different concentrations of Rg3. Results HNE-1 cells could form TLSs and mosaic vessels when mix-cultured with CRL-2480 on Matrigel. Rg3 could inhibit the TLS formation of HNE-1 cells. Mter 24-hour culture in medium with Rg3 at concentrations of 0, 50, 100 and 200 μg/ml, the number of TLSs were 75.50 ±6.86,55.00 ± 11.92, 39.75 ± 7.93 and 24.50 ± 6.25, respectively, which were negatively correlated with the concentrations of Rg3 ( r = - 0. 928;P ＜ 0. 01 ). After 48 hours of culture, the expressions of VEGF significantly declined by IHC test with results as 0. 19 ±0. 03, 0. 13 ±0. 02, 0. 11 ±0. 01, and 0.08 ±0.01, respectively, which were negatively correlated with the concentrations of Rg3 ( r = -0. 911;P ＜0.01). The expressions of VEGF also gradually decreased as revealed by Western blot test, with corresponding results as 119.49, 111.51, 86.45, and 38.29. All of the tests showed significantly declined results in the group at the concentration of 200 μg/ml Rg3. Conclusion Rg3 can inhibit the vasculogenic mimicry of HNE-1 cells, and the possible mechanism might be associated with the down-regulation of VEGF protein expression in HNE-1 cells.