咪唑诱导的人子宫内膜癌细胞HEC-1B空泡化现象研究

Study on the Vacuolation of Imidazole-Treated Human Endometrial Carcinoma Cell Line HEC-1B

咪唑作为一种弱碱性物质,处理动物子宫内膜细胞后,可诱导其产生空泡化现象,进而影响其生殖生理功能。本研究使用人子宫内膜癌细胞HEC-1B作为模型,使用咪唑处理导致细胞空泡化后探讨空泡化产生的生理机制。通过基因芯片分析、荧光定量PCR检测、激光共聚焦显微镜观察等方法进行研究。结果表明:咪唑处理人子宫内膜癌细胞HEC-1B后,空泡型质子泵(V-ATPase)亚基V0D2基因表达量上调。使用荧光定量PCR验证芯片结果,V-ATPase亚基V0D2 mRNA表达量有所上调。对V-ATPase另外一个亚基V1E1基因进行荧光定量PCR,其mRNA表达量也有所上调。使用V-ATPase抑制剂巴弗洛霉素A1和咪唑共同处理人子宫内膜癌细胞HEC-1B后,通过吖啶橙染色,激光共聚焦显微镜观察,咪唑诱导的空泡被显著抑制。上述结果证明了V-ATPase在咪唑诱导的细胞空泡化中的重要作用。

As a kind of weak bases, imidazole cause vacuolation of animal endometrial cells, and affect the physiological and reproductive functions. Our study discuss the mechanism of vacuolation after treatment with imidazole through experimental model of human endometrial carcinoma cell line HEC-lB. We study through Genechip research, fluorescence quantitative PCR and observation of laser confocal microscopy. Results ： Genechip research results show that gene of subunit VOD2 of human vacuolar type H＋-ATPase （V-ATPase） had an increased expression after the treatment with imidazole on human endometrial carcinoma cell line HEC-lB. Genechip research results were verified by fluorescence quantitative PCR. The expression of the mRNA of subunit VOD2 of human V-ATPase increased.Another subunit V1E1 of human V-ATPase also had an increased expression by fluorescence quantitative PCR. Bafilomycin A1, an inhibitor of V-ATPase, significantly inhibited the vacuolation caused by imidazole. The phenomenon was observed by laser confocal microscopy after staining human endometrial carcinoma cell line HEC-1B with Acridine orange. These results suggest that V-ATPase plays an important role in cell vacuolation caused by imidazole.