MCL-1反义寡核苷酸转染恶性黑素瘤细胞的光镜和电镜观察

Observation on the Antisense MCL-1 Transfected Malignant Melanoma Cells by Light Microscope and Electron Microscope

目的通过转染MCL-1反义寡核苷酸观察转染后细胞形态的变化,以研究MCL-1基因的功能。方法采用阳离子脂质体转染MCL-1反义寡核苷酸,以特异性阻断MCL-1表达。用光镜观察转染前后细胞显微结构变化及免疫细胞化学染色结果;以电镜观察转染前后细胞微观形态的变化。实验中设立正义序列寡核苷酸和无义序列寡核苷酸作为对照寡核苷酸。结果光镜下可见到转染后细胞变圆,胞内出现空泡。免疫组化结果显示细胞内阳性染色明显减少。电镜观察显示转染后细胞内出现空泡,细胞膜突起消失及出现凋亡细胞。结论MCL-1基因对恶性黑素瘤细胞形态有很大的影响。本研究为阐明MCL-1基因的功能提供了较好的形态学证据。

To observe the morphological alteration on antisense MCL-1 transfected malignant melanoma cells and further study the function of MCL-1. Methods MCL-1 ASODN was transfected into melanoma cells with lipofectamine 2000, and the cells were observed under light microscope and eletron microsope. The sense oligonucleotide and the nonsense oligonucleotide were used as the control oligonucleotides. Results Both light microscope and electron microscope revealed abnormal mophology of antisense MCL-1 transfected malignant melanoma cells. They turned to round, and vacuolus developed in the cytoplasm by microscope. Immunohistochemistry revealed decreased MCL-1 expression levels. The electron microscopic observation indicated decreased microvilli,swelling mitochondria. Apoptosis was observed, too. Conclusion MCL-1 has great effect on the morphology of melanoma cells. What we have done provides useful morphological proof in the study for the function in MCL-1.