Zmiany ekspresji podjednostek kanału BK w komórkach linii U87-MG wt i U87-MG ∆αBK po traktowaniu czynnikami wywołującymi starzenie komórkowe

The experiment was performed on U87-MG WT and U87-MG ΔαBK cells, both control and treated with 300 µM H₂O₂ or 20 nM doxorubicin. Gene expression was analyzed by PCR using primers specific for genes encoding different isoforms of the BK channel α subunit, including DEC, STREX, VYR, and the glioma-specific isoform gBK. GAPDH was used as an internal control.Under the same experimental conditions, PCR reactions were performed to determine the expression levels of the β and γ subunits of the BK channel.The PCR products were visualized on a 2% agarose gel stained with Midori Green Advance DNA Staining (MG04).

本实验以U87-MG野生型（U87-MG WT）与U87-MG αBK缺失型（U87-MG ΔαBK）细胞为研究对象，对两类细胞均设置对照组及分别经300 μM过氧化氢（H₂O₂）、20 nM多柔比星处理的实验组。采用聚合酶链式反应（PCR），使用靶向编码BK通道α亚基不同剪接异构体的特异性引物，分析编码BK通道α亚基各异构体（包括DEC、STREX、VYR以及胶质瘤特异性亚型gBK）的基因表达水平。以甘油醛-3-磷酸脱氢酶（GAPDH）作为内参基因。在相同实验条件下，通过PCR反应检测BK通道β、γ亚基的表达水平。PCR产物在经Midori Green Advance DNA染色剂（MG04）染色的2%琼脂糖凝胶上进行可视化分析。