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目的研究反义整合素β6基因对结肠癌细胞生长增殖的作用。方法先构建反义β6基因的表达载体,然后转染结肠癌细胞,使其稳定表达。应用 RT-PCR 检测细胞中β6mRNA 的表达量;应用流式细胞技术检测细胞表面β6蛋白的表达量;检测分析各组细胞与纤连蛋白结合力的改变;检测分析各组细胞增殖能力的改变;将转染细胞及正常细胞分别种植于裸鼠皮下,观察细胞的成瘤能力。将实验组与对照组的检测结果进行比较。结果与对照组相比,转染了反义β6基因的结肠癌细胞,其β6mRNA 的量及细胞表面β6蛋白的表达量(30%)均明显减少(P<0.01);其与纤连蛋白的结合力降低;其生长及增殖的能力较对照组细胞明显减弱;其在裸鼠皮下的成瘤能力明显减弱(P<0.01)。结论反义β6基因可以在转录和翻译水平明显抑制β6基因的表达,从而抑制结肠癌细胞的生长增殖,证实了整合素β6对结肠癌细胞的生长增殖具有重要的调控作用。
Objective To study the effect of antisense integrin β6 gene on the proliferation and proliferation of colon cancer cells. Methods The antisense β6 gene expression vector was constructed and then transfected into colon cancer cells for stable expression. The expression of β6 mRNA in cells was detected by RT-PCR. The expression of β6 protein on the cell surface was detected by flow cytometry. The changes of the binding capacity of fibronectin were detected and analyzed. Transfected cells and normal cells were implanted in nude mice subcutaneously to observe the tumorigenic ability of the cells. The experimental group and control group test results were compared. Results Compared with the control group, the amount of β6 mRNA and the expression of β6 protein (30%) in the colon cancer cells transfected with antisense β6 gene were significantly decreased (P <0.01). Compared with the control group, The binding capacity decreased; its ability of growth and proliferation was significantly weaker than that of the control group; its tumorigenic ability in nude mice subcutaneously decreased (P <0.01). CONCLUSION: Antisense β6 gene can significantly inhibit the expression of β6 gene at transcription and translation levels, thereby inhibiting the growth of colon cancer cells. It is confirmed that integrin β6 plays an important regulatory role in the growth and proliferation of colon cancer cells.