Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method

查看全文 作　　者：Zhiwei [1,3]Wu;Qing [2]Ye;Yimei [1]Huang;Yating [2]Lin;Mengmeng [1]Zheng;Xiaoyan [1]Wang;Shusen [1]Xie;Juqiang [1]Lin 机构地区：[1]MOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial Key Laboratory for Photonics Technology Fujian Normal University Fuzhou 350007,P.R.China;[2]Department of Otolaryngology Provincial Clinical College of Fujian Medical University Fujian Provincial Hospital Fuzhou 350001,P.R.China;[3]School of Physical and Information Engineering Quanzhou Normal University Quanzhou 362000,P.R.China 出　　处：《Journal of Innovative Optical Health Sciences》索引2019年第6期,共11页 基　　金：supported by the National Key Basic Research Program of China(No.2015CB352006);the National Natural Science Foundation of China(Nos.61335011,61775037 and 61475036);Program for Changjiang Scholars and Innovative Research Team in University(No.IRT 15R10);Natural Science Foundation of Fujian Province of China(Nos.2019J01270 and 2017J01844);the High level Joint Research and Construction Program of Fujian Provincial Hospital,and Special Funds of the Central Government Guiding Local Science and Technology Development(2017L3009),China. 摘　　要：Latent mermbrane protein 1(LMP1)is known as an oncoprotein in nasopharyngeal carcinoma(NPC)cells,which is considered to have a strong association with growth,invasion and metastasis of NPC cells through lipid rafts.Met hy-A-cyclodextrin(M3CD)can disrupt lipid rafts through cholesterol depletion.In this study,we revealed that MICD induced apoptosis in two kinds of NPC cells including CNE1 cells,a LMP1 negative nasopharyngeal squamous carcinoma cell line,and CNE1-LMP1 cells,a LMP1-positive nasopharyngeal squamous carcinoma cell line.Furthermore,the impact of MBCD on LMP1 was investigated by fuorescence resonance energy transfer(FRET)method in NPC cells.Synchronized tempo spatial and spectral detection of LMP1/LMP1 interaction were performed using fluorescence microscope and spectrograph.FRET efficiency indicated that LMP1/LMP1 interaction gradully enhanced after M9CD treatment.MTT assays showed that M8CD caused strong cytotoxicity in CNE1 cells,but cauused relatively weaker cytotoxicity in CNE1-LMP1 cells,which indicated that LMP1 may regulate sensitivity of NPC cells to MBCD.Then,detection of cleaved caspase-3 in two kinds of NPC cells indicated that LMP1 may inhibit activation of caspase 3.These results strongly suggested that MBCD can induce apoptosis of NPC cells,but enhancing of LMP1/LMP1 interaction may likely resist apoptosis through inhibiting cleavage of caspase-3. 关 键 词：Fluorescence resonance energy transfer(FRET) nasopharyngeal carcinoma(NPC) latent mermbrane protein 1(LMP1) lipid rafts