PEI介导PKC-α反义核酸对肝癌SMMC-7721细胞的增殖抑制作用

Proliferative inhibition effects of PKC-α antisense oligodeoxynuleotides mediated by polyethylenimine on SMMC-7721 cell lines

目的:研究聚乙烯亚胺(PEI)-PKC-α反义脱氧核寡酸(ASODN)对人肝癌SMMC-7721细胞的增殖抑制作用。方法:RT-PCR检测肝癌SMMC-7721细胞、HepG2细胞PKC-α mRNA表达的差异,细胞增殖抑制实验检测PEI和ASODN混合的最佳质量比,WST法和克隆形成抑制实验检测细胞增殖抑制作用,免疫荧光检测PKC-α的表达水平。结果:SSMC-7721细胞PKC-α mRNA表达相对较高,PEI和ASODN的质量比为0.75/1时是PEI转染反义核酸的合适比例,对SMMC-7721细胞具有明显的增殖克隆抑制作用,且呈剂量-效应关系;ASODN和PEI-ASODN对SMMC-7721的IC50分别为16.6μmol/L和0.58μmol/L;PEI-ASODN能够有效抑制PKC-α蛋白的生物合成。结论:PEI-ASODN能显著抑制SMMC-7721细胞增殖和克隆形成,下调PKC-α蛋白的表达。

Aim:To investigate the proliferative inhibition effect of antisense oligonucleotides(ASODN) targeting for protein kinase C alpha(PKC-α) mediated by polyethyleneimine(PEI) on hepatocelluar carcinoma SMMC-7721 cells. Methods: The difference of PKC-α mRNA expression on hepatocelluar carcinoma SMMC-7721 cells and HepG2 cells was detected by reverse transcription-polymerase chain reaction(RT-PCR).The best qualitative ratio of PEI/ASODN was determined by proliferative inhibition on SMMC-7721 cells.PKC-α ASODN were transfected with PEI,the proliferation and the clone formation of SMMC-7721 cells were detected by WST method and clone formation assay,respectively,and the expression of PKC-α was analyzed by immunofluorescence test.Results: The expression of PKC-α mRNA was higher in SMMC-7721 cells than that in HepG2 cells,the best qualitative ratio of PEI /ASODN on SMMC-7721 cells was at 0.75/1.Compare with the control group,PEI group,the proliferation and clone formation rates of SMMC-7721 cells treated with PEI-ASODN were significantly inhibited(P<0.05),and the inhibitive effect was much more significant when the dose of PEI-ASODN increased.The IC50 of ASODN on SMMC-7721 cells' proliferation was 16.6 μmol/L and the IC50 of PEI-ASODN on SMMC-7721 cells was 0.58 μmol/L.And the expression of PKC-α protein in SMMC-7721 cells treated with PEI-ASODN was significantly lower than those in the control group detected by immunofluorescence test.Conclusion:The PKC-α ASODN mediated by PEI can inhibit the proliferation and clone formation in SMMC-7721 cells significantly and can down-regulate the expression of PKC-α protein.