变异IκBα抑制肝癌细胞株HCC9204中NF-κB的活性及侵袭转移

Nuclear Factor KappaB Activity and Cell Migration Metastasis of HCC9204 inhibited by Mutated Inhibitor Kappa Alpha

目的探讨重组腺病毒变异IκBα(mIκBα)对HCC9204中NF-κB活性和转移相关能力的影响。方法利用有限稀释法测定病毒滴度,扩增AdmIκBα的滴度为1.252×109pfu/mL,以MOI 20、35、50的AdIκBαm感染HCC9204细胞;Western印迹检测细胞NF-κB(p65)的表达,EMSA法检测NF-κB的活性;以细胞粘附人工重组基底膜实验检测HCC9204细胞的粘附能力;用Transwell小室法检测HCC9204的侵袭和运动能力。结果MOI=20时,NF-κB的活性、P65蛋白表达开始下降,在MOI=50时降低到最低水平,AdIκBαm对NF-κB表达的抑制呈剂量依赖性,并且HCC9204细胞的细胞黏附、运动和侵袭转移能力逐渐降低。结论重组腺病毒变异IκBa抑制HCC9204中NF-κB的活性及体外细胞的浸润转移能力。

Objective To investigate the effects of mutant inhibitory kappaB α （mI kB α） gene transfection into human hepatocellular carcinoma cell lines（HCC9204） on migration metastasis and its mechanisms involved in this effect.Methods To test the virus titer in 293 cells with limited dilution method ,the titer ofAdI kB α M is 1.252 × 10^9pfu/mL.And infected HCC9204cells with MOI 20, 35, 50,respectively.The expression of p65 was investigated by Western blot,the activity of NF- kB was detected through EMSA,the effects of I kB α m on the adhesion,migration,metastasis assay of HCC9204 cells were investigated by Transwell and Matrigel methods.Results The activity of NF- κB and the expression of p65 protein began to decreace when MOI was 10,and the lowest when MOI was 50.The cells through the PVPF in Invasion and Migration Assay and the cells through PVPF membrance in Migration Assay were lower than that of non-infected cells.Conclusion The expression of mutant I κB α gene（mIκBα）can inhibit the activity of NF- κB, which can desease migration, Invasion and Migration.