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目的检测八聚体结合蛋白-4(octamer-binding protein-4,OCT4)基因在不同分化程度的胃癌细胞株(MKN-28、SGC-7901、BGC-823)及正常胃黏膜细胞株GES-1中的表达情况,研究OCT4基因表达水平与胃癌细胞分化程度和侵袭力的相关性。方法 RT-PCR确定GES-1、MKN-28、SGC-7901、BGC-823中OCT4基因的表达水平,OCT4siRNA转染BGC-823细胞,分别通过荧光定量PCR和Western blot检测干扰OCT4基因表达的效果,并利用Transwell小室法研究OCT4在BGC-823细胞侵袭力中的作用。结果正常人胃黏膜细胞中未检测到OCT4的表达,人胃癌细胞系分化程度越低,OCT4基因表达量越高,低分化人胃癌细胞系BGC-823中OCT4基因表达最高,转染OCT4siRNA的BGC-823细胞内的OCT4mRNA和OCT4蛋白表达量明显下降(P<0.05)。干扰胃癌细胞BG-823中OCT4基因的表达,穿膜细胞数量减少(P<0.05),侵袭力下降。结论 OCT4基因表达量与胃癌细胞的分化程度有关,OCT4基因可能在胃癌细胞的分化中起重要作用,影响胃癌细胞的侵袭能力,其表达程度有望成为胃癌患者恶性度预测的参考指标之一。
Objective To detect the expression of OCT4 gene in gastric cancer cell lines (MKN-28, SGC-7901, BGC-823) and normal gastric mucosal cell line GES-1 In the expression of OCT4 gene expression levels and gastric cancer cell differentiation and invasiveness of the correlation. The expression of OCT4 gene in GES-1, MKN-28, SGC-7901 and BGC-823 cells was determined by RT-PCR. OCT4 siRNA was transfected into BGC-823 cells and the expression of OCT4 gene was detected by real-time PCR and Western blot respectively Transwell chamber method was used to study the role of OCT4 in invasiveness of BGC-823 cells. Results OCT4 expression was not detected in normal human gastric mucosa cells. The lower the differentiation of human gastric cancer cell lines was, the higher the expression of OCT4 gene was. The highest expression of OCT4 gene was in BGC-823 cell line, and the expression of OCT4 siRNA in BGC The expression of OCT4 mRNA and OCT4 protein in -823 cells decreased significantly (P <0.05). Interfered the expression of OCT4 gene in BG-823 gastric cancer cells, decreased the number of transmembrane cells (P <0.05) and decreased invasiveness. Conclusion The expression of OCT4 gene is related to the differentiation of gastric cancer cells. OCT4 gene may play an important role in the differentiation of gastric cancer cells and affect the invasion ability of gastric cancer cells. The expression of OCT4 gene may be one of the reference indexes for predicting the malignancy of gastric cancer patients.