Li Xian, Li Shanshan, Wang Xinxin, Zhao Surong, Liu Hao
College of Pharmacy, Bengbu Medical College, Bengbu 233030, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2018 Dec 30;38(12):1409-1414. doi: 10.12122/j.issn.1673-4254.2018.12.02.
To study the effect of knocking down fascin on cervical cancer cell proliferation and tumorigenicity in nude mice.
Cervical cancer CaSki cells were infected with a lentiviral vector carrying fascin siRNA or with a negative control lentivirus, and fascin mRNA 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.
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 ( < 0.05). Fascin knockdown resulted in significantly reduced proliferation of CaSki cells ( < 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.
Knocking down fascin can inhibit the growth and tumorigenicity of cervical cancer cells in nude mice.
研究敲低肌动蛋白丝束蛋白对子宫颈癌细胞增殖及裸鼠致瘤性的影响。
用携带丝束蛋白小干扰RNA的慢病毒载体或阴性对照慢病毒感染子宫颈癌CaSki细胞,采用实时定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测细胞中丝束蛋白的信使核糖核酸(mRNA)和蛋白表达。采用噻唑蓝(MTT)比色法检测敲低丝束蛋白后CaSki细胞的增殖情况。将转染丝束蛋白小干扰RNA或对照慢病毒载体的CaSki细胞以及未转染的CaSki细胞皮下接种于裸鼠,测量移植瘤的体积和重量;采用蛋白质免疫印迹法检测移植瘤中增殖细胞核抗原(PCNA)、生存素、细胞周期蛋白依赖性激酶4(CDK4)及p21蛋白的表达。
携带丝束蛋白小干扰RNA的慢病毒载体感染CaSki细胞后,而非阴性对照载体,可使细胞中丝束蛋白mRNA和蛋白表达水平显著降低(<0.05)。敲低丝束蛋白可导致CaSki细胞增殖显著减少(<0.05)。与对照小鼠相比,接种敲低丝束蛋白的CaSki细胞的裸鼠移植瘤体积和重量减小,PCNA、生存素和CDK4水平降低,移植瘤中p21蛋白表达增加。阴性对照慢病毒不影响CaSki细胞在裸鼠中的增殖或致瘤性,也不影响移植瘤中PCNA、生存素、CDK4或p21蛋白的表达水平。
敲低丝束蛋白可抑制子宫颈癌细胞在裸鼠中的生长及致瘤性。