Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method
Latent membrane 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. Methyl-β-cyclodextrin (MβCD) can disrupt lipid rafts through cholesterol deplet...
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| Format: | Article |
| Language: | English |
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World Scientific Publishing
2019-11-01
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| Series: | Journal of Innovative Optical Health Sciences |
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| Online Access: | http://www.worldscientific.com/doi/pdf/10.1142/S1793545819500196 |
| _version_ | 1811215245027311616 |
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| author | Zhiwei Wu Qing Ye Yimei Huang Yating Lin Mengmeng Zheng Xiaoyan Wang Shusen Xie Juqiang Lin |
| author_facet | Zhiwei Wu Qing Ye Yimei Huang Yating Lin Mengmeng Zheng Xiaoyan Wang Shusen Xie Juqiang Lin |
| author_sort | Zhiwei Wu |
| collection | DOAJ |
| description | Latent membrane 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. Methyl-β-cyclodextrin (MβCD) can disrupt lipid rafts through cholesterol depletion. In this study, we revealed that MβCD 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 MβCD on LMP1 was investigated by fluorescence 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 gradually enhanced after MβCD treatment. MTT assays showed that MβCD caused strong cytotoxicity in CNE1 cells, but caused relatively weaker cytotoxicity in CNE1-LMP1 cells, which indicated that LMP1 may regulate sensitivity of NPC cells to MβCD. 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 MβCD can induce apoptosis of NPC cells, but enhancing of LMP1/LMP1 interaction may likely resist apoptosis through inhibiting cleavage of caspase-3. |
| first_indexed | 2024-04-12T06:18:06Z |
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| institution | Directory Open Access Journal |
| issn | 1793-5458 1793-7205 |
| language | English |
| last_indexed | 2024-04-12T06:18:06Z |
| publishDate | 2019-11-01 |
| publisher | World Scientific Publishing |
| record_format | Article |
| series | Journal of Innovative Optical Health Sciences |
| spelling | doaj.art-745f5fc2062b40118537a0c32ce727f32022-12-22T03:44:23ZengWorld Scientific PublishingJournal of Innovative Optical Health Sciences1793-54581793-72052019-11-011261950019-11950019-1110.1142/S179354581950019610.1142/S1793545819500196Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET methodZhiwei Wu0Qing Ye1Yimei Huang2Yating Lin3Mengmeng Zheng4Xiaoyan Wang5Shusen Xie6Juqiang Lin7MOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial, Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, P. R. ChinaDepartment of Otolaryngology, Provincial Clinical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou 350001, P. R. ChinaMOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial, Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, P. R. ChinaDepartment of Otolaryngology, Provincial Clinical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou 350001, P. R. ChinaMOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial, Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, P. R. ChinaDepartment of Otolaryngology, Provincial Clinical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou 350001, P. R. ChinaMOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial, Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, P. R. ChinaMOE Key Laboratory of Optoelectronic Science and Technology for Medicine and Fujian Provincial, Key Laboratory for Photonics Technology, Fujian Normal University, Fuzhou 350007, P. R. ChinaLatent membrane 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. Methyl-β-cyclodextrin (MβCD) can disrupt lipid rafts through cholesterol depletion. In this study, we revealed that MβCD 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 MβCD on LMP1 was investigated by fluorescence 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 gradually enhanced after MβCD treatment. MTT assays showed that MβCD caused strong cytotoxicity in CNE1 cells, but caused relatively weaker cytotoxicity in CNE1-LMP1 cells, which indicated that LMP1 may regulate sensitivity of NPC cells to MβCD. 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 MβCD can induce apoptosis of NPC cells, but enhancing of LMP1/LMP1 interaction may likely resist apoptosis through inhibiting cleavage of caspase-3.http://www.worldscientific.com/doi/pdf/10.1142/S1793545819500196fluorescence resonance energy transfer (fret)nasopharyngeal carcinoma (npc)latent membrane protein 1 (lmp1)lipid rafts |
| spellingShingle | Zhiwei Wu Qing Ye Yimei Huang Yating Lin Mengmeng Zheng Xiaoyan Wang Shusen Xie Juqiang Lin Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method Journal of Innovative Optical Health Sciences fluorescence resonance energy transfer (fret) nasopharyngeal carcinoma (npc) latent membrane protein 1 (lmp1) lipid rafts |
| title | Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method |
| title_full | Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method |
| title_fullStr | Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method |
| title_full_unstemmed | Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method |
| title_short | Real-time detection of LMP1/LMP1 interaction in MβCD-induced apoptosis of nasopharyngeal carcinoma cells using FRET method |
| title_sort | real time detection of lmp1 lmp1 interaction in mβcd induced apoptosis of nasopharyngeal carcinoma cells using fret method |
| topic | fluorescence resonance energy transfer (fret) nasopharyngeal carcinoma (npc) latent membrane protein 1 (lmp1) lipid rafts |
| url | http://www.worldscientific.com/doi/pdf/10.1142/S1793545819500196 |
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