安徽医药
安徽醫藥
안휘의약
ANHUI MEDICAL AND PHARMACEUTICAL JOURNAL
2014年
11期
2036-2039
,共4页
蛋白磷酸酶2A(PP2A)%调节亚基B56ε%细胞转化%DNA损伤修复
蛋白燐痠酶2A(PP2A)%調節亞基B56ε%細胞轉化%DNA損傷脩複
단백린산매2A(PP2A)%조절아기B56ε%세포전화%DNA손상수복
protein phosphatase 2A(PP2A)%regulatory B56εsubunit%cell transformation%DNA damage and repair
目的：探讨蛋白磷酸酶2A-B56ε亚基(PP2A B56ε,由PPP2R5E基因编码)的表达在苯并芘诱导细胞转化过程中对DNA损伤修复功能的影响。方法实时荧光定量-PCR(QRT-PCR)检测BEAS-2B细胞(人正常肺上皮细胞)、和BEAS-2BT细胞(苯并芘诱导恶变的人肺上皮细胞)中PPP2R5E的mRNA转录水平；免疫印迹法(Western Blot)检测细胞中 B56ε编码蛋白的水平；中性单细胞凝胶电泳实验(彗星实验,SCGE)检测喜树碱( camptothecin,CPT)处理的细胞DNA断裂损伤及修复情况；免疫印迹法检测 CPT 处理诱导γH2AX(磷酸化组蛋白 H2AX)水平。结果与 BEAS-2B 细胞相比较,BEAS-2BT 细胞中PPP2R5E其mRNA水平及B56ε蛋白水平均显著高表达(P<0.05)。与BEAS-2B细胞相比,经CPT处理恢复后,BEAS-2BT细胞尾距显著降低(P<0.05)。表明BEAS-2BT细胞中H2AX的磷酸化水平明显降低。结论在苯并芘诱导细胞转化过程中PP2A B56ε通过介导γH2AX去磷酸化影响DNA损伤修复功能。
目的：探討蛋白燐痠酶2A-B56ε亞基(PP2A B56ε,由PPP2R5E基因編碼)的錶達在苯併芘誘導細胞轉化過程中對DNA損傷脩複功能的影響。方法實時熒光定量-PCR(QRT-PCR)檢測BEAS-2B細胞(人正常肺上皮細胞)、和BEAS-2BT細胞(苯併芘誘導噁變的人肺上皮細胞)中PPP2R5E的mRNA轉錄水平；免疫印跡法(Western Blot)檢測細胞中 B56ε編碼蛋白的水平；中性單細胞凝膠電泳實驗(彗星實驗,SCGE)檢測喜樹堿( camptothecin,CPT)處理的細胞DNA斷裂損傷及脩複情況；免疫印跡法檢測 CPT 處理誘導γH2AX(燐痠化組蛋白 H2AX)水平。結果與 BEAS-2B 細胞相比較,BEAS-2BT 細胞中PPP2R5E其mRNA水平及B56ε蛋白水平均顯著高錶達(P<0.05)。與BEAS-2B細胞相比,經CPT處理恢複後,BEAS-2BT細胞尾距顯著降低(P<0.05)。錶明BEAS-2BT細胞中H2AX的燐痠化水平明顯降低。結論在苯併芘誘導細胞轉化過程中PP2A B56ε通過介導γH2AX去燐痠化影響DNA損傷脩複功能。
목적：탐토단백린산매2A-B56ε아기(PP2A B56ε,유PPP2R5E기인편마)적표체재분병비유도세포전화과정중대DNA손상수복공능적영향。방법실시형광정량-PCR(QRT-PCR)검측BEAS-2B세포(인정상폐상피세포)、화BEAS-2BT세포(분병비유도악변적인폐상피세포)중PPP2R5E적mRNA전록수평；면역인적법(Western Blot)검측세포중 B56ε편마단백적수평；중성단세포응효전영실험(혜성실험,SCGE)검측희수감( camptothecin,CPT)처리적세포DNA단렬손상급수복정황；면역인적법검측 CPT 처리유도γH2AX(린산화조단백 H2AX)수평。결과여 BEAS-2B 세포상비교,BEAS-2BT 세포중PPP2R5E기mRNA수평급B56ε단백수평균현저고표체(P<0.05)。여BEAS-2B세포상비,경CPT처리회복후,BEAS-2BT세포미거현저강저(P<0.05)。표명BEAS-2BT세포중H2AX적린산화수평명현강저。결론재분병비유도세포전화과정중PP2A B56ε통과개도γH2AX거린산화영향DNA손상수복공능。
Objective To investigate protein phosphatase 2A-B56εsubunit(PP2A B56ε,encoded by the gene PPP2R5E) expression to DNA damage and repair functions during benzopyrene induced cell transformation. Methods Real-time quantitative-PCR ( QRT-PCR) was used to detect the BEAS-2B cells (normal human lung epithelial cells) and BEAS-2BT cells (benzo(α)pyrene induced malignant human lung epithelial cells) mRNA transcript levels in PPP2R5E. SCGE assay was adopted to evaluate their genotoxicity. Western blot was used to detect theγH2AX(phosphorylated histone H2AX ) level. Results Compared with the BEAS-2B cells,the mRNA and pro-tein encoded by PPP2R5E and its expression levels were significantly higher(P<0. 05). SCGE analysis showed that,compared with BEAS-2B cells,BEAS-2BT cells tail was significantly reduced (P<0. 05). Compared with BEAS-2B cells,γH2AX expression levels of BEAS-2BT cells were significantly higher(P<0. 05). Conclusions In the process of benzo(α)pyrene induced cell transformation, PP2A B56εaffects DNA damage and repair by mediating throughγH2AX dephosphorylation.
