聚乙烯亚胺-整合素配体复合物介导bcl-2反义核酸对结肠癌细胞的作用

目的:观察聚乙烯亚胺(PEI) -整合素蛋白的配体(Arg -Gly -Asp ,RGD)介导的bcl - 2反义核酸(antisenseoligodeoxynucleotide ,ASODN)对结肠癌细胞caco - 2的作用效果。方法:将阳离子复合物jetPEI-RGD与bcl- 2反义核酸混合形成ASODN -PEI-RGD三聚体复合物,转染caco - 2细胞。用台盼蓝拒染法计数活细胞,观察ASODN -PEI-RGD对caco - 2细胞的生长抑制作用,用流式细胞仪检测细胞的亚二倍体百分率,Heochst332 5 8染色观察细胞凋亡。结果:与ASODN对照组和jetPEI-RGD对照组相比,ASODN -PEI -RGD能够明显抑制caco - 2细胞增殖和诱导细胞凋亡(P<0 0 5 ) ,呈剂量和时间-效应关系。ASODN -PEI-RGD作用4 8h ,荧光染色可见大量的caco - 2细胞核固缩、核碎裂。结论:jetPEI-RGD介导的bcl- 2反义核酸能抑制caco - 2细胞增殖和诱导细胞凋亡。

Effect of bcl-2 antisense oligodeoxynucleotide mediated by jetPEI-RGD on colon carcinoma cell line caco-2

Aim: To observe if bcl-2 antisense oligodeoxynucleotide (ASODN) mediated by jetPEI-RGD improved effect on colon carcinoma cell line caco-2. Methods: Mixing (bcl-2) ASODN with jetPEI-RGD to get ASODN-PEI-RGD, treating with caco-2 cell lines. Inhibition of cells proliferation after treatment with ASODN-PEI-RGD was inspected by trypan blue exclusion assay. Apoptotic changes were observed by fluorescence microscopy and flow cytometry. Results: Proliferation of caco-2 cell surpressed significantly after treatment with ASODN-PEI-RGD compared with control groups (P<0.05) and showed dose-dependent and time-dependent inhibitory effects. After treatment with ASODN-PEI-RGD for 48 hour, many cells stained with Hoechst33258 exhibited apoptotic morphology such as cell shrinkage, nuclear condensation and nuclear fragmentation under fluorescence microscope. Conclusion:jetPEI-RGD can efficiently transfect ASODN for inhibiting proliferation effect and inducing apoptosis on caco-2 cells in vitro.