重组人血管内皮抑制素对食管癌细胞增殖、凋亡及侵袭作用的机制研究

Effects of recombinant human endostatin on proliferation,apoptosis and invasion of human esophageal cancer cells

目的研究重组人血管内皮抑制素对食管癌细胞(KYSE150)体外增殖、凋亡及侵袭蛋白表达的的影响。方法将食管癌细胞随机分为对照组和低、中、高3个剂量实验组。对照组和低、中、高3个剂量实验组分别给予0,100,200,400μg·mL^(-1)重组人血管内皮抑制素,持续培养48 h。用噻唑蓝(MTT)法检测食管癌细胞增殖抑制率。用Annexin V-FITC/PI法检测细胞凋亡情况,用逆转录聚合酶链式反应(RT-PCR)检测Notch1 mRNA和Notch2 mRNA的表达情况,用蛋白免疫印迹法检测血管内皮生长因子-C(VEGF-C)蛋白的表达情况。结果流式细胞仪分析结果提示,重组人血管内皮抑制素促进人食管癌细胞凋亡。低、中、高剂量实验组Notch1 mRNA、Notch2 mRNA表达均显著低于对照组(均P<0.05),VEGF-C蛋白表达均显著低于对照组(均P<0.05)。低、中、高3个剂量实验组细胞凋亡率分别为(5.63±1.54)%,(8.14±2.31)%,(15.31±2.07)%,对照组为(2.11±0.89)%,差异均有统计学意义(均P<0.05)。结论重组人内皮抑制素对食管癌细胞凋亡具有显著促进作用,对癌细胞增殖有抑制作用,这可能是Notch信号通路影响的表现。

Objective To study the effects of recombinant human endostatin on proliferation,apoptosis and invasion protein expression of esophageal cancer cells（ KYSE150） in vitro. Methods The esophageal cancer cells were randomly divided into control group and low,middle and high dose experimental groups. Control group and low,middle and high dose experimental groups were given 0,100,200,400 μg·mL^-1 recombinant human endostatin,and cultured for 48 h. The inhibition rate on proliferation of esophageal cancer cells was detected by MTT assay.The apoptosis of the cells was detected by Annexin V-FITC/PI,and the expressions of Notch1 mRNA and Notch2 mRNA were detected by reverse transcription polymerase chain reaction（ RT-PCR）,and the expression of vascular endothelial growth factor（ VEGF-C） protein was detected by protein immunoblotting. Results The results of flow cytometry showed that recombinant human endostatin could promote the apoptosis of human esophageal cancer cells. The expressions of Notch1 mRNA and Notch2 mRNA in low,middle and high dose experimental groups were significantly lower than those in control group（ all P〈0. 05）,and the expression of VEGF-C in experimental groups was significantly lower than that in control group（ all P〈0. 05）.The apoptotic rates in low,middle and high dose experimental groups were（ 5. 63 ± 1. 54） %,（ 8. 14 ± 2. 31） %,（ 15. 31 ± 2. 07） %,had significant difference with that in control group,which was（ 2. 11 ± 0. 89） %（ P〈0. 05）.Conclusion Recombinant human endostatin has significant promotion on the apoptosis of esophageal cancer cells and inhibits the proliferation of cancer cells,which may be a manifestation of the effect of Notch signaling pathway.