靶向干扰前列腺源性ETS因子促进HT29细胞的增殖与侵袭

摘要

Objective:Colorectal cancer is one of the common gastrointestinal tumors.Recent studies have shown that, the expression of PDEF can promote the differentiation of Secretory progenitor cells to goblet cells in the intestinal tissue.Therefore the oc-currence of colorectal cancer may be related to expression of PDEF.In this study,we tried to investigate the effects of proliferation and invasion after interference targeting prostate-derived ETS factor in colorectal cell lines HT29.Methods: HT29 cells were transiently transfected with PDEF shRNA plasmids and blank control plasmid via cathodolyte liposome transfection method.By fluorescence microscopy,RT-PCR,Western blot technique to detect the expression of PDEF mRNA and protein in normal control group,blank control group,shRNA group.The proliferation and invasion ability of HT29 cells after transfection were assessed by MTT assay and Transwell invasion assay respectively.Results: Green fluorescent protein was observed in blank control plasmid group and shRNA plasmid group.Western blot showed the reduced PDEF protein expression compared with normal control group and blank control group.Interference PDEF gene expression can significantly promote the proliferation of HT29 cells (P<0.05).The ability of cell invasion in interference group was significantly higher than the normal control group and blank control group after 48h ( P<0.05).Conclusion:Interference PDEF in HT29 cells can promote cell proliferation and invasion.%目的：结直肠癌是常见的消化道肿瘤之一，研究发现在结直肠组织，前列腺源性ETS因子（ Prostate-derived Ets factor，PDEF）的表达可以促进分泌性祖细胞向杯状细胞分化，因此结直肠癌的发生可能与PDEF的表达有关。本研究旨在探讨靶向干扰前列腺源性ETS因子对结直肠癌细胞株HT29增殖与侵袭的影响。方法：阳离子脂质体法将PDEF干扰质粒和空白对照空质粒瞬时转染HT29细胞，通过荧光显微镜、RT-PCR、Western blot技术测定正常对照组、空白对照组和shRNA组中PDEF mRNA和蛋白的表达情况；MTT法检测HT29细胞的增殖情况；采用Transwell迁移实验观察细胞侵袭能力。结果：干扰质粒、空质粒成功转染HT29细胞，通过荧光显微镜可以观察到绿色荧光蛋白的表达；Western blot结果可见shRNA组PDEF蛋白的表达较正常对照组和空白对照组降低；MTT法检测发现干扰PDEF基因的表达能够明显促进HT29细胞的增殖（ P＜0.05）；48 h后，shRNA组细胞侵袭能力明显强于正常对照组和空白对照组（P＜0.05）。结论：在HT29细胞中干扰PDEF的表达，可以促进细胞的增殖与侵袭。