敲低Fascin抑制宫颈癌细胞增殖及裸鼠的成瘤能力

Knocking down fascin inhibits cervical cancer cell proliferation and tumorigenesis in nude mice

目的研究敲低Fascin对宫颈癌细胞增殖及裸鼠成瘤能力的影响。方法 Fascin siRNA慢病毒(干扰组)和阴性对照慢病毒(阴性组)感染宫颈癌CaSki细胞,用qRT-PCR和Western blot方法检测Fascin mRNA以及蛋白表达水平评价干扰效果。MTT方法测定Fascin敲低对CaSki细胞增殖能力的影响。取对照组、阴性组、干扰组CaSki细胞接种至裸鼠皮下,建立裸鼠移植瘤模型,测定移植瘤生长体积和质量,Western blot方法检测移植瘤组织中增殖细胞核抗原(PCNA)、survivin、细胞周期依赖性蛋白激酶4(CDK4)、p21蛋白水平。结果 Fascin siRNA慢病毒感染后的CaSki细胞中Fascin mRNA和蛋白水平低于没有感染的CaSki细胞(P<0.05)。阴性对照慢病毒感染对CaSki细胞中Fascin mRNA和蛋白水平没有影响。敲低Fascin后的CaSki细胞增殖能力明显降低(P<0.05)。干扰组CaSki接种的裸鼠移植瘤体积和质量也降低,移植瘤组织中PCNA、survivin、CDK4蛋白水平降低,p21蛋白水平升高。阴性组CaSki细胞接种对细胞增殖、裸鼠成瘤能力及移植瘤中PCNA、survivin、CDK4、p21蛋白水平没有影响。结论敲低Fascin能够抑制宫颈癌细胞生长和裸鼠成瘤能力。

Objective To study the effect of knocking down fascin on cervical cancer cell proliferation and tumorigenicity in nude mice. Methods Cervical cancer CaSki cells were infected with a lentiviral vector carrying fascin siRNA or with a negative control lentivirus, and fascin m RNA and protein expressions in the cells were detected using qRT-PCR and Western blotting.MTT assay was used to determine the proliferation of CaSki cells with fascin knockdown. CaSki cells transfected with fascin siRNA or the control lentiviral vector and non-transfected CaSki cells were inoculated subcutaneously in nude mice, and the volume and weight of the transplanted tumor were measured; Western blotting was used to detect the expressions of proliferating cell nuclear antigen(PCNA), survivin, cyclin dependent kinase 4(CDK4) and p21 proteins in the tumor xenograft. Results Infection with the lentiviral vector carrying fascin siRNA, but not the negative control vector, caused significant reductions in the expression levels of fascin mRNA and protein in CaSki cells(P<0.05). Fascin knockdown resulted in significantly reduced proliferation of CaSki cells in vitro(P<0.05). The nude mice inoculated with CaSki cells with fascin knockdown showed reduced tumor volume and weight, lowered levels of PCNA, survivin and CDK4, and increased expression of p21 protein in the tumor xenograft compared with the control mice. The negative control lentivirus did not affect the proliferation or tumorigenicity of CaSki cells in nude mice or the expression levels of PCNA, survivin, CDK4 or p21 proteins in the xenografts. Conclusion Knocking down fascin can inhibit the growth and tumorigenicity of cervical cancer cells in nude mice.