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目的 :观察骨桥蛋白(osteopontin,OPN)小干扰RNA稳定转染人胃癌细胞SGC-7901后,细胞增殖、凋亡及迁移能力的变化并探讨其可能的分子机制。方法:用脂质体法分别将OPNsiRNA-pcDNATM6.2及空载体质粒pcDNATM6.2转染人胃癌细胞SGC-7901,稻瘟素筛选,克隆环挑取细胞克隆,用Western blot及RT-PCR技术筛选阳性克隆,并检测稳转细胞中OPN、趋化因子受体4(CXCR4)、基质金属蛋白酶2(MMP2)的表达,应用MTT法、流式细胞仪、细胞迁移试验分别检测转染细胞的增殖、凋亡及迁移能力。结果:OPN siRNA稳定转染细胞SGC-7901后,稳转细胞中CXCR4和MMP2表达量下降;细胞的增殖能力(与转染空载体质粒比较)降低、细胞凋亡率显著高于对照组,转染细胞的迁移能力明显降低。结论:OPN siRNA使胃癌细胞增殖和迁移能力降低、细胞凋亡增加;提示OPN可能通过SDF-1/CXCR4轴及MMP2参与的信号通路调节肿瘤细胞生长及转移。
OBJECTIVE: To observe the changes of proliferation, apoptosis and migration of human gastric cancer cell line SGC-7901 after osteopontin (OPN) small interfering RNA was stably transfected into human gastric cancer cell line SGC-7901 and explore its possible molecular mechanism. METHODS: OPNsiRNA-pcDNATM6.2 and empty vector plasmid pcDNATM6.2 were transfected into human gastric cancer cell line SGC-7901 by lipofectamine respectively. The cell clones were picked out by cloning loop and cloned by Western blot and RT-PCR The positive clones were screened and the expressions of OPN, CXCR4 and MMP2 were detected by MTT assay. The expressions of OPN, CXCR4 and MMP2 were detected by flow cytometry and MTT assay respectively Proliferation, apoptosis and migration ability. Results: The expression of CXCR4 and MMP2 in stable transfected cells was decreased after OPN siRNA was stably transfected into SGC-7901 cells. The proliferation ability of OPN siRNA transfected with SGC-7901 cells was lower than that of transfected empty vector and the apoptosis rate was significantly higher than that of control group Dye cell migration was significantly reduced. CONCLUSION: OPN siRNA can reduce the proliferation and migration of gastric cancer cells and increase the apoptosis of cells. It suggests that OPN may regulate the growth and metastasis of tumor cells through the signal pathways involved in SDF-1 / CXCR4 axis and MMP2.