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目的:探讨纳米脂质体槲皮素(nLQ)诱导人食管癌Eca-9706细胞逆转化及凋亡的效应及其机制。方法:采用旋转蒸发法制备以氯仿和DMSO为溶剂的脂质体槲皮素,将其超声波破碎并经80nm滤膜过滤制成nLQ,不同浓度(10、20、40、80和100μmol/L)作用于Eca-9706细胞,用MTT法检测各组Eca-9706细胞的生长抑制率。分别取Eca-9706细胞,分为nLQ组(加40μmol/LnLQ)和对照组。TUNEL法检测2组细胞凋亡率;免疫细胞化学/免疫印迹法检测2组处理48h后Eca-9706细胞CyclinD1、PTEN、c-Met、VEGF、组蛋白去乙酰化酶(HDAC)1及NF-κB表达的变化。结果:各组Eca-9706细胞生长抑制率(F分别为128.041、142.683和231.054,P<0.001)和2组细胞凋亡率(t=94.770,P<0.001)比较,差异均有统计学意义。PTEN免疫反应性和印记信号增强(t分别为5.352和4.308,P均<0.05),Cyclin D1、c-Met、VEGF、HDAC1和NF-κB的免疫反应性减弱(t分别为4.006、9.184、13.853、4.698和3.575,P均<0.05),其印迹信号亦减弱(t分别为3.827、6.399、7.868、4.695和3.406,P均<0.05);以上各细胞蛋白的免疫细胞和免疫印迹信号之间呈正相关(rs分别为0.952、0.915、0.927、0.842、0.879和0.855,P均<0.05)。结论:nLQ可抑制Eca-9706细胞生长及增殖,逆转Eca-9706细胞PTEN的低表达和c-Met及VEGF的高表达,并通过抑制HDAC1和NF-κB及激活PTEN表达的HDAC抑制信号途径而诱导细胞凋亡。
Objective: To investigate the effect and mechanism of nano-liposome quercetin (nLQ) on the reversal and apoptosis of human esophageal cancer Eca-9706 cells. Methods: The liposome quercetin with chloroform and DMSO as the solvent was prepared by rotary evaporation method. The liposome quercetin was disrupted by ultrasonic wave and filtered through 80nm filter to obtain nLQ. Different concentrations (10, 20, 40, 80 and 100μmol / L) Eca-9706 cells treated with Eca-9706 cells by MTT assay. Eca-9706 cells were divided into nLQ group (plus 40μmol / LnLQ) and control group. The apoptosis rates of Eca-9706 cells in two groups were detected by TUNEL method. The expressions of CyclinD1, PTEN, c-Met, VEGF, HDAC-1 and NF- κB expression changes. Results: The growth inhibition rates of Eca-9706 cells in each group were 128.041, 142.683 and 231.054, respectively (P <0.001), and the apoptosis rates of two groups were statistically significant (t = 94.770, P <0.001). PTEN immunoreactivity and imprinting signals were enhanced (t = 5.352 and 4.308 respectively, P <0.05), while the immunoreactivity of Cyclin D1, c-Met, VEGF, HDAC1 and NF-κB decreased (t = 4.006,9.184,13.853 , 4.698 and 3.575, all P <0.05), and their imprinted signals also weakened (t = 3.827,6.399,7.868,4.695 and 3.406 respectively, P <0.05). There was positive correlation between the immunocytes and immunoblots (Rs = 0.952, 0.915, 0.927, 0.842, 0.879 and 0.855 respectively, all P <0.05). CONCLUSION: nLQ can inhibit the growth and proliferation of Eca-9706 cells and reverse the low expression of PTEN and the high expression of c-Met and VEGF in Eca-9706 cells. By inhibiting HDAC1 and NF-κB and activating HDAC signal pathways, Induces apoptosis.