CAJ | 학술논문

目的研究以慢病毒为载体靶向人乳头瘤病毒16型(HPV16)E6基因的短发夹状干扰RNA(shRNA)对宫颈癌Caski细胞侵袭能力的影响.方法将靶向HPV16型E6的shRNA表达序列(干扰组)和无义表达(无义干扰组)克隆到慢病毒工作质粒中包装成病毒颗粒,感染宫颈癌Caski细胞.观察绿色荧光阳性细胞的比例以了解慢病毒对Caski细胞的感染效率;用RT-PCR法检测shRNA作用后3组Caski细胞HPV16型E6 mRNA含量的变化;Western blot法检测E6蛋白表达的变化;Transwell小室法检测肿瘤细胞侵袭能力的变化.结果慢病毒对Caski细胞的最佳每个细胞中的病毒颗粒数目(MOI)为2.5;与无义干扰组相比,干扰组细胞中HPV16E6的mRNA含量减少70%,蛋白表达水平降低63%,Caski细胞的侵袭能力显著下降.结论慢病毒携带的shRNA能干扰宫颈癌Caski细胞中HPV16型E6的表达,并抑制癌细胞的侵袭能力.
목적 연구이만병독위재체파향인유두류병독16형(HPV16)E6기인적단발협상간우RNA(shRNA)대궁경암Caski세포침습능력적영향.방법 장파향HPV16형E6적shRNA표체서렬(간우조)화무의표체(무의간우조)극륭도만병독공작질립중포장성병독과립,감염궁경암Caski세포.관찰록색형광양성세포적비례이료해만병독대Caski세포적감염효솔;용RT-PCR법검측shRNA작용후3조Caski세포HPV16형E6 mRNA함량적변화;Western blot법검측E6단백표체적변화;Transwell소실법검측종류세포침습능력적변화.결과 만병독대Caski세포적최가매개세포중적병독과립수목(MOI)위2.5;여무의간우조상비,간우조세포중HPV16E6적mRNA함량감소70%,단백표체수평강저63%,Caski세포적침습능력현저하강.결론 만병독휴대적shRNA능간우궁경암Caski세포중HPV16형E6적표체,병억제암세포적침습능력.
Objective To investigate the influence of lentivirus mediated short hairpin RNA(shRNA)target to human papilloma virus(HPV)16 E6 on invasive ability of cervical cancer Caski cells.Methods Lentivirus was produced after shRNA target to human papilloma virus(HPV)16 E6 and to nonsense was cloned to lentivirus work vector.Infection ratio was assessed by assay of EGFP positive cells of Caski.Total mRNA of E6 was determined by RT-PCR after Caski cells were infected by lentivirus.The change of E6 protein expression was analyzed by Western blot.The invasive ability of Caski cells was assayed employing Transwell.Results The optimal MOI (Multiplicity of infection)of lentivirus to Caski was 2.5.Total mRNA and protein of E6 were decreased (by 70%and 63%)in interfering group compared with control group.The invasive ability of Caski cells also reduced after infected by lentivirus.Conclusion shRNA mediated by lentivirus can inhibit expression of HPV16 E6 and invasive ability of cervical cancer cells.