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目的研究抗癌一号KAI1(又称CD82)基因对骨髓瘤细胞生物学特性的影响及机制。方法将携带KAI1基因克隆至重组腺病毒,制备Ad5-KAI1重组腺病毒,并感染骨髓瘤细胞SKO-007。Ad5-GFP作为阴性对照,未感染细胞为空白对照,利用CCK-8和AnnexinⅤ-PI等方法分别检测不同时间点(24 h,48 h)的细胞活率和凋亡水平。另外,利用W estern印迹方法检测不同组别细胞表达ERK及其磷酸化水平。结果 Ad5-KAI1组细胞活率抑制水平和凋亡水平明显高于对照组,且剪切型胱天蛋白酶(caspase)3、ERK磷酸化水平在实验组高表达,而抑制ERK磷酸化则导致凋亡进一步增加。结论 KAI1显著抑制骨髓瘤细胞增殖,并诱导其caspase依赖的凋亡;KAI1在促进凋亡的同时,其应激性诱导了ERK磷酸化,后者又对细胞具有保护作用。
Objective To study the effect of anti-cancer KAI1 (aka CD82) gene on the biological characteristics of myeloma cells and its mechanism. Methods The KAI1 gene was cloned into the recombinant adenovirus and the Ad5-KAI1 recombinant adenovirus was prepared and infected with myeloma SKO-007. Ad5-GFP as a negative control, uninfected cells as a blank control, CCK-8 and Annexin Ⅴ-PI were used to detect cell viability and apoptosis levels at different time points (24 h, 48 h). In addition, Western blotting was used to detect the expression of ERK and its phosphorylation in different groups of cells. Results The cell viability and apoptosis levels in Ad5-KAI1 group were significantly higher than those in control group, and the phosphorylation level of ERK phosphorylation was significantly higher in the experimental group than in the control group, while the phosphorylation of ERK resulted in apoptosis Death increased further. Conclusions KAI1 significantly inhibits the proliferation of myeloma cells and induces its caspase-dependent apoptosis. While KAI1 promotes apoptosis, KAI1 induces phosphorylation of ERK, which in turn has a protective effect on the cells.