stathmin基因反义寡核苷酸对Eca109细胞系生长的抑制作用

Inhibitory effects of ASODN of stathmin gene on cultured Eca109 cell lines

目的:研究stathmin基因反义寡核苷酸(ASODN)对Eca109细胞增殖及stathmin基因表达的抑制作用,探讨ASODN用于食管癌基因治疗的可行性。方法:实验设ASODN转染组、SODN转染组和未转染组。分别应用stathmin ASODN、SODN转染食管癌Eca109细胞。倒置显微镜观察转染细胞的形态变化,绘制细胞生长曲线,MTT法检测细胞增殖能力,RT-PCR方法检测stathmin基因的表达,采用流式细胞仪分析细胞增殖周期。结果:stathmin ASODN转染Eca109细胞后,胞体凝缩,增殖能力减弱,细胞生长及目的基因表达明显受抑(P<0.05),其抑制作用具有序列特异性。细胞分裂阻滞在分裂期的中期,G2/M期细胞数由(12.2±1.0)%升至(36.5±5.8)%。结论:stathmin基因反义寡核苷酸对Eca109细胞增殖及stathmin基因表达具有明显抑制作用,有望成为食管癌基因治疗药物。

Aim ： To investigate the inhibition of stathmin oligodeoxynucleotides（ASODN） on the expression of stath- min gene and the proliferation of Eca109 cells and to explore the possibility of the ASODN as drug for esophageal cancer gene therapy. Methods：Eca109 cells were allocated into 3 groups：ASODN transfected group, SODN transfected group and non-transfected group. After the transfection, the cell proliferation of Eca109 cell lines was observed with invert microscope, growth curve and MTT methods. The expression of stathmin mRNA was detected by RT-PCR. The cell cycle was analyzed by flow cytometer. Results：After being blocked by ASODN, the cell bodies become condensation, the growth of cells was repressed, and the proliferation ability and the expression of stathmin gene was inhibited obviously（ P 〈 0.05）. The inhibition was specific in sequence. The cell division was blocked in metaphase, and the Eca109 cells in G2/M phase in- creased from （12.2 ± 1. 0）% to （36.5 ± 5.8）%. Conclusion： The stathmin ASODN can inhibit the proliferation of Eca109 cells by blocking the expression of stathmin gene and may be a new gene drug for treatment of esophageal cancer.