磁性纳米复合物对人肝癌细胞增殖能力的影响

Effect of magnetic nanocomposites on proliferation ability of human hepatoma carcinoma cells

目的探讨磁性纳米复合物对人肝细胞癌(肝癌)细胞(Hep G2细胞株)增殖能力的影响。方法以聚乙烯亚胺包被的磁性纳米复合物(PEI-SPIO)作为基因载体复合富含亮氨酸重复单位的G蛋白耦联受体(LGR)5-小干扰RNA(siRNA),待细胞融合达60%时转染Hep G2细胞建立PEI组,等量的单纯LGR5-siRNA转染Hep G2建立SI组,另设未转染对照(Ctrl)组。采用MRI T2扫描检测纳米复合物进入细胞的效率,细胞计数试剂盒(CCK)-8实验检测细胞增殖抑制率,逆转录聚合酶链反应(RT-PCR)检测细胞LGR5的信使核糖核酸(mRNA)表达水平,蛋白印迹法检测LGR5、cyclin D1蛋白表达。结果 PEI组Hep G2细胞的MRI T2信号明显减低。与Ctrl组相比,PEI组Hep G2细胞的细胞增殖抑制率明显升高,LGR5 mRNA的相对表达量和LGR5、cyclin D1蛋白相对表达量均明显降低(均为P<0.05),而SI组细胞的相应指标差异无统计学意义(均为P>0.05)。结论磁性纳米复合物PEI-SPIO复合LGR5-siRNA可有效转染Hep G2细胞,其机制可能是通过下调cyclin D1表达水平抑制人肝癌Hep G2细胞的增殖能力。

Objective To investigate the effect of magnetic nanocomposites on proliferation ability of human hepatoma carcinoma （HCC）cells （HepG2 cell line）.Methods Leucine-rich repeat-containing G protein-coupled receptor （LGR） 5-small interfering ribonucleic acid （siRNA ） was composited with polyethylenimine wrapped superparamagnetic iron oxide nanoparticle （PEI-SPIO）as the gene vector.PEI group was established by transfecting HepG2 cells when cell fusion reached 60% and SI group was established by transfecting HepG2 cells with equivalent simple LGR5-siRNA.Control （Ctrl）group was also established without transfecting.The efficiency of nanocomposites entering cells was scanned with MRI T2.The inhibition rate of cell proliferation was detected by （cell count kit,CCK）-8 assay.The expression level of messenger ribonucleic acid （mRNA）in LGR5 of cells was detected by reverse transcriptase polymerase chain reaction （RT-PCR）and the protein expressions of LGR5 and cyclin D1 were detected by western blotting.Results MRI T2 signal of HepG2 cells in PEI group decreased significantly.Compared with Ctrl group,the inhibition rate of cell proliferation of HepG2 cells in PEI group was significantly increased.The relative expression of LGR5 mRNA and the relative expression of LGR5 and cyclin D1 protein were both significantly decreased （all in P 〈0.05）,while the corresponding indexes of the cells in SI group had no statistical significance （all in P〉0.05 ）.Conclusions Magnetic nanocomposites PEI-SPIO composited with LGR5-siRNA may effectively transfect HepG2 cells.Its mechanism may take effect through down-regulating the expression of cyclin D1 to inhibit the proliferation ability of hepatocellular carcinoma HepG2 cells.