受体介导的c-myc反义核酸抑制肝癌Bel-7402细胞增殖的机制

目的探讨半乳糖受体介导的c-myc反义寡核苷酸(antisenseoligodeoxynucleotide,ASODN)抑制肝癌Bel-7402细胞增殖的机制。方法c-mycASODN与半乳糖(galactose,Gal)-聚乙烯亚胺(poly-ethyleneimine,PEI)相作用形成Gal-PEI-ASODN复合物,作用于人肝癌Bel-7402细胞,通过流式细胞仪检测细胞周期、AnnexinV-FITC和碘化丙啶(PropidiumIodide,PI)双染色、DNA电泳实验观察Gal-PEI-ASODN对Bel-7402细胞的作用。结果采用流式细胞仪检测细胞周期,细胞对照组、Gal-PEI对照组、ASODN对照组,细胞增殖指数分别为35.04%、33.95%、32.90%,Gal-PEI-ASODN组细胞增殖指数为23.65%,与细胞对照组相比,sub-G1期+G0/G1期细胞总数从64.03%增加到76.74%,S期+G2/M期的细胞从35.04%减少为23.65%,Gal-PEI-ASODN组细胞增殖指数下降11.39%,差异显著(P<0.01)。采用细胞凋亡检测试剂AnnexinV-FITC/P...

Mechanism of c-myc Antisense Oligodeoxynucleotide Mediated by Receptor on the Proliferative Inhibition of Bel-7402 Cells

Objective To observe the mechanism of c-myc antisense oligodeoxynucleotide mediated by galactose receptor on the proliferative inhibition of hepatocarcinoma Bel-7402 cells.Methods c-myc ASODN mixed with Galactose(Gal)- polyethyleneimine(PEI) reagent forming Gal-PEI-ASODN complex,then incubated with Bel-7402 cells for 48h,using Propidium Iodide(PI) dying,AnnexinV-FITC and PI double staining,DNA gel electrophoresis to detected the proliferative inhibition mechanism of Gal-PEI-ASODN on Bel-7402 cells.Results Cell cycle was determined by flow cytometry.In the control group,Gal-PEI group and ASODN group,cell proliferative index was found to be 35.04% , 33.95% , 32.90% ,respectively.However,cell proliferative index for Gal-PEI-ASODN group was found to be 23.65% ,decreased by 11.39% compared to the control group(P< 0.01 ).Necrotic cell percentage was determined using AnnexinV-FITC/ PI double staining test.The results showed cell apoptotic rate of 2.77% ,cell necrotic rate of 3.06% and the normal cell percentage was detected as 94.13% in the control group;cell apoptotic rate below 4%,cell necrotic rate below 6.8% were detected in both ASODN group and Gal-PEI group; cell apoptotic rate of 6.5% ,cell necrotic rate of 15.9% and the normal cell percentage of 76% were determined in Gal-PEI-ASODN group.There was no DNA ladder morphology,on the contrary,there was DNA smear morphology using DNA gel electrophoresis in Gal-PEI-ASODN group.Conclusion The mechanism by which Gal-PEI-ASODN inhibits cell proliferation maybe that it can restrain the change from G_ 0 /G_ 1 period to S period in cell cycle and induces cell necrosis.