摘要
目的探讨CXCR7-shRNA慢病毒表达载体靶向抑制CXCR7的表达对人肝癌细胞增殖及侵袭能力的影响。方法利用RNA干扰技术,构建CXCR7的shRNA慢病毒表达载体,转染人肺转移肝癌细胞株HCCLM3,分别通过RT-PCR和Western blot检测HCCLM3细胞中CXCR7 mRNA和蛋白质表达水平的变化;然后通过MTT法考察CXCR7沉默对HCCLM3细胞增殖侵袭能力的影响,通过体外粘附实验和Transwell小室法分别考察CXCR7沉默对HCCLM3细胞粘附及侵袭能力的影响。结果与空白对照组比较,转染CXCR7-shRNA慢病毒载体的HCCLM3细胞中CXCR7的mRNA及蛋白水平表达均明显降低(P<0.01),SDF-1诱导的HCCLM3细胞的增殖能力显著下降(P<0.05),同时HCCLM3细胞的粘附及侵袭能力也明显受到CXCR7-shRNA的抑制(P<0.01)。结论靶向沉默CXCR7能显著抑制结肝癌细胞的增殖和侵袭能力,为肝癌的治疗提供一个潜在的作用靶点。
Objective To explore the effects of CXCR7 knock-down by CXCR7-shRNA lentiviral vector on the proliferation and invasion of human hepatoma carcinoma cells in vitro. Methods CXCR7- shRNA lentiviral vector was transfected into heptatocellular carcinoma HCCLM3 cells. The changes in mRNA and protein expression of CXCR7 in the transfected cells were investigated using real- time PCR and Western blotting, respectively, and MTT assay was employed to assess the cell proliferation changes. In vitro adhesion assay and transwell chamber test were used to observe the adhesion and invasiveness of HCCLM3 cells, respectively. Results Transfection of HCCLM3 cells with CXCR7- shRNA lentiviral vector resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P0.01) and obvious suppression of the cell proliferative activity (P0.05). CXCR7-shRNA also significantly suppressed the invasiveness and adhesion of HCCLM3 cells (P 0.01). Conclusion CXCR7 knock- down can significantly inhibit the proliferation and invasiveness of human hepatoma carcinoma cells in vitro, suggesting the value of CXCR7 as a potential target for hepatoma carcinoma therapy.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第7期994-998,共5页
Journal of Southern Medical University
基金
国家自然科学基金(81200917
81201702)
四川省教育厅资助科研项目(10ZC092)
重庆市自然科学基金项目(cstc2012jjA10139)
重庆大学"生物流变科学与技术"教育部重点实验室访问学者基金(CQKLBST-2012-003)~~