槲皮素PLGA-TPGS纳米粒在肝癌细胞中的摄取及其细胞抑制率的研究

Investigation on cellular uptake of quercetin-loaded PLGA-TPGS nanoparticles to liver cancer cells and its cell inhibition ratio

目的:考察槲皮素(quercetin,QT)/香豆素-6(coumarin-6,C6)乙交酯丙交酯共聚物-维生素E聚乙二醇1000琥珀酸酯(polylactide-co-glycolide-D-α-tocopheryl polyethylene glycol 1000 succinate,PLGA-TPGS)纳米粒(QT/C6-loaded PLGATPGS nanoparticles,QCPTN)体外分别被人肝癌细胞株HepG2和小鼠腹水型高淋巴道转移肝癌细胞HCa-F的摄取情况,以及研究槲皮素PLGA-TPGS纳米粒(QT-loaded PLGA-TPGS nanoparticles,QPTN)体外对HepG2细胞的生长抑制率。方法:用激光共聚焦显微镜考察2种肝癌细胞HepG2和HCa-F对QCPTN的体外细胞摄取情况。采用WST-1法体外研究QPTN和槲皮素PLGA纳米粒(QT-loaded PLGA nanoparticles,QPN)对HepG2细胞生长的抑制性。试验分为6组,分别为阴性对照组、空白纳米粒(empty PLGA-TPGS nanoparticles,EPTN)组、氟尿嘧啶溶液(fluorouracil solutions,FS)组、槲皮素溶液(quercetin solutions,QTS)组、QPN组、QPTN组,分别在培育24、48、72 h加入WST-1后在酶标仪上450 nm下测定吸光度值,计算对HepG2细胞的生长抑制率。结果:在2种不同类型肝癌细胞摄取试验中,激光共聚焦显微镜镜下均可见显绿色荧光的QCPTN分布在2种肝癌细胞的细胞核周围,表明QCPTN可被HepG2和HCa-F细胞分别摄取进入细胞内。体外肝癌细胞生长抑制率试验结果表明,自制材料PLGA-TPGS对HepG2细胞生长无明显的抑制性;QPN和QPTN的体外对HepG2细胞生长抑制率有浓度依赖性和时间依赖性;QPTN对HepG2细胞的抑制率大于QPN、QTS和FS。结论:QCPTN通过细胞摄取作用可将模型药物QT和荧光标记物C6同时带入HepG2和HCa-F细胞内,QPTN体外显示对HepG2细胞生长具有较强的抑制性,与QPN、QTS和FS相比,具有较好的抑制肝癌细胞生长的作用。

OBJECTIVE To investigate the cellular uptake of quercetin（QT）/coumarin-6（C6）-loaded polylactide-co-glycolide-D-α-tocopheryl polyethylene glycol 1000 succinate（PLGA-TPGS）nanoparticles（QCPTN）in the human liver cancer cell line HepG2 and the ascitic hepatocarcinoma cell strain,which exhibited high metastatic potential in the lymphatic system（HCaF cells）,and investigate the in vitro cell inhibition ratio of QT-loaded PLGA-TPGS nanoparticles（QPTN）to HepG2 cells.METHODS QCPTN＇s cellular uptakes by HepG2 and HCa-F cells were investigated using a confocal laser scanning microscope（CLSM）.In vitro cell inhibition ratios of QPTN and QT-loaded PLGA nanoparticles（QPN）to HepG2 cells were assessed by the WST-1 assay.The cells were incubated with QPTN,QPN,quercetin solutions（QTS）,fluorouracil solutions（FS）and empty PLGA-TPGS nanoparticles（EPTN）for 24,48 and 72 h,and then joined by WST-1,respectively.The absorption was determined at a wavelength of 450 nm by a microplate reader,and cell inhibition ratio was calculated.RESULTSIn the study of cellular uptake in the two kinds of liver cancer cells,the entire area of both cancer cells,especially the cytoplasm exhibited the strong green fluorescence,which proved that QCPTN had been internalized into liver cancer cells efficiently and rapidly.In vitro cell inhibition ratio results showed that PLGA-TPGS showed no obvious cell inhibition effect to HepG2 cells.QPTN and QPN exhibited dose-dependent,time-dependent in the in vitro cell inhibition test.QPTN exhibited higher cell inhibition effects than QPN,RS and FS.CONCLUSION QCPTN can internalize and deliver both model drug QT and the fluorescent marker C6 into liver cancer cells HepG2 and HCa-F efficiently by cellular uptake.Cell inhibition test in vitro demonstrates that QPTN,which exhibits higher inhibition effects to HepG2 cells,show better cell inhibition effects compared with QPN,QTS and FS.