CAJ | 학술논문

目的观察洛铂对人胆管癌RBE细胞微丝F-肌动蛋白(F-actin)和α-微管蛋白(α-tubulin)作用.方法体外培养胆管癌RBE细胞,分别用不同剂量洛铂处理,鬼笔环肽-异硫氰酸荧光素(Phalloidin-FITC)和4',6-二脒基-2-苯基吲哚(DAPI)双染后激光共聚焦显微镜观察微丝蛋白F-actin结构变化,免疫组织化学染色观察α-tubulin结构变化,实时定量聚合酶链反应(Real-time PCR)检测α-tubulin mRNA转录水平变化.结果激光共聚焦显微镜及免疫组织化学染色法观察到洛铂作用下,胆管癌RBE细胞微丝蛋白F-actin与微管蛋白α-tubulin均发生解聚和重排,而α-tubulin染色加深(P＜0.05),α-tubulin mRNA/β-actin结果:5.0 mg/L组:2.08 ±0.13;25.0 mg/L组:1.91±0.46(P ＜0.05),证实洛铂使胆管癌RBE细胞α-tubulin转录水平增加.结论微丝、微管的解聚和重排在洛铂对胆管癌RBE细胞诱导凋亡过程中起重要作用.
목적 관찰락박대인담관암RBE세포미사F-기동단백(F-actin)화α-미관단백(α-tubulin)작용.방법 체외배양담관암RBE세포,분별용불동제량락박처리,귀필배태-이류청산형광소(Phalloidin-FITC)화4',6-이미기-2-분기신타(DAPI)쌍염후격광공취초현미경관찰미사단백F-actin결구변화,면역조직화학염색관찰α-tubulin결구변화,실시정량취합매련반응(Real-time PCR)검측α-tubulin mRNA전록수평변화.결과 격광공취초현미경급면역조직화학염색법관찰도락박작용하,담관암RBE세포미사단백F-actin여미관단백α-tubulin균발생해취화중배,이α-tubulin염색가심(P＜0.05),α-tubulin mRNA/β-actin결과:5.0 mg/L조:2.08 ±0.13;25.0 mg/L조:1.91±0.46(P ＜0.05),증실락박사담관암RBE세포α-tubulin전록수평증가.결론 미사、미관적해취화중배재락박대담관암RBE세포유도조망과정중기중요작용.
Objective To investigate the F-actin and α-tubulin cytoskeleton changes in cholangiocarcinoma cell line RBE after treatment with Lobaplatin.Methods The RBE cells were cultured in vitro,followed by treatment with various concentrations of lobaplatin for 24 h.The changes of F-actin cytoskeleton were scanned by confocal laser scanning microscopy (CLSM) after staining with phalloidin-fluoresceine isothiocyanate (FITC) and DAPI.Immunohistochemistry and real-time polymerase chain reaction (Real-time PCR) were used to observe α-tubulin cytoskeleton changes and α-tubulin mRNA transcript levels respectively.Results CLSM and immunohistochemistry showed that both the F-actin and the α-tubulin cytoskeleton of RBE cells had disorganization and rearrangement after the treatment with lobaplatin.The staining of α-tubulin was getting deep (P ＜ 0.05),and the Real-time PCR determined α-tubulin mRNA/β-actin ratio in 5.0 mg/L and 25.0 mg/L lobaplatin-treated groups was 2.08 ±0.13 and 1.91 ± 0.46 respectively (P ＜ 0.05).Conclusion The changes of microfilaments and microtubules in RBE cells play a positive role in the apoptosis induced by lobaplatin.