siRNA对恶性黑色素瘤A375细胞中HOXB7基因的干涉作用研究
Effect of siRNA Targeting the HOXB7 Gene in Human Malignant Melanoma A375 Cells
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摘要: 目的:观察靶向siRNA抑制恶性黑色素瘤细胞株A375同源盒基因HOXB7表达后,对相应生长因子bFGF表达水平及A375细胞生物学特性的影响。方法:根据HOXB7基因设计3条序列特异性的siRNA(si-HOXB7-1,si-HOXB7-2,si-HOXB7-3),在脂质体Lipofectamine2000的介导下转染A375细胞,采用RT-PCR和FCM法来观察HOXB7及bFGF的基因沉默效应。MTT法检测A375细胞体外增殖活性。结果:3条siRNA均能不同程度地下调A375细胞HOXB7表达水平,第3条序列能更有效地封闭HOXB7基因,而空转组与对照组相比差异无统计学意义(P>0.05)。结论:靶向HOXB7基因的siRNA可有效沉默HOXB7基因及其调控的基因bFGF的表达,并控制恶性黑色素瘤细胞的生长。Abstract: Objective: To observe the influence of small interfering RNA (siRNA) targeted against the HOXB7 gene on expression of HOXB7 and bFGF and other biologic characteristics in the A375 malignant melanoma cell line. Methods: Three sequence-specific siRNA's (si-HOXB7-1, si-HOXB72, si-HOXB7-3) were designed based on the HOXB7 gene sequence and were transfected into A375 cells by lipofectamine 2000 induction. Gene silencing effects of HOXB7 and bFGF were observed using RT-PCR and flow cytometry. MTT colorimetry was used to detect cell proliferative activity in vitro. Results: The expression of HOXB7 in A375 cells could be downregulated in varying degrees by the 3 siRNA's. The 3rd siRNA can most effectively block expression of the HOXB7 gene, so there was no significant difference between the free-running group and the control group (P>0.05). Conclusion: siRNA targeting the HOXB7 gene can effectively downregulate the expression of HOXB7 and its downstream gene bFGF and repress proliferation of A375 cells.
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