蛋白組學(xué)分析放射抗拒性鼻咽癌細(xì)胞差異表達(dá)蛋白
發(fā)布時間:2019-02-18 15:12
【摘要】:目的篩選同一來源放射敏感性不同的鼻咽癌細(xì)胞蛋白組學(xué)差異表達(dá),探討鼻咽癌放射抗拒機(jī)制。方法建立放射敏感性不同的人鼻咽癌細(xì)胞株,采用雙向凝膠電泳技術(shù)和圖像分析軟件PDQuest分離和分析不同階段細(xì)胞間的差異蛋白質(zhì)點(diǎn),采用LC-MS/MS質(zhì)譜分析系統(tǒng)鑒定差異蛋白質(zhì)點(diǎn),采用Gene Oncology Annotation將已知差異蛋白質(zhì)進(jìn)行分類。部分篩選蛋白質(zhì)采用Western blot進(jìn)行驗(yàn)證。結(jié)果 CNE-2和CNE-2R細(xì)胞有差異候選差異蛋白質(zhì)27個,其中11個蛋白質(zhì)在放射抗拒CNE-2R細(xì)胞中表達(dá)下調(diào),16個上調(diào)。其中的4個:4-3-3σ、GADD45、PRKDC和HSP27用Western blot的方法得到了進(jìn)一步確認(rèn)。結(jié)論 CNE-2細(xì)胞受射線反復(fù)照射產(chǎn)生放射抗拒性細(xì)胞,在蛋白組學(xué)水平上發(fā)生了某些突變,并將這些突變穩(wěn)定的遺傳下來,從而表現(xiàn)出放射抗拒性的表型;通過調(diào)控其中與放射抗拒性產(chǎn)生相關(guān)的某些蛋白表達(dá),就有可能調(diào)控細(xì)胞的放射敏感。4-3-3σ、GADD45、PRKDC和HSP27是預(yù)測NPC放療敏感性的潛在基因標(biāo)志物,它們表達(dá)失調(diào)可能參與NPC放射抗拒性的產(chǎn)生。
[Abstract]:Objective to screen the proteomic differential expression of nasopharyngeal carcinoma cells with different radiosensitivity from the same source, and to explore the mechanism of radioresistance in nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell lines with different radiosensitivity were isolated and analyzed by two-dimensional gel electrophoresis and image analysis software PDQuest. Differential protein spots were identified by LC-MS/MS mass spectrometry and known differential proteins were classified by Gene Oncology Annotation. Some of the selected proteins were verified by Western blot. Results there were 27 candidate proteins in CNE-2 and CNE-2R cells, 11 of which were down-regulated and 16 up-regulated in radiation-resistant CNE-2R cells. Four of them: 4-3-3 蟽, GADD45,PRKDC and HSP27 were confirmed by Western blot method. Conclusion Radio-resistant cells were produced by repeated irradiation of CNE-2 cells, and some mutations occurred at the proteomic level, and these mutations were inherited stably, thus showing the phenotype of radioresistance. By regulating the expression of some proteins associated with radioresistance, it is possible to regulate the radiosensitivity of cells. 4-3-3 蟽, GADD45,PRKDC and HSP27 are potential gene markers for predicting the radiosensitivity of NPC. They may be involved in the production of radioresistance in NPC.
【作者單位】: 西安交通大學(xué)醫(yī)學(xué)院第二附屬醫(yī)院腫瘤病院放療科;
【基金】:國家自然科學(xué)基金資助項(xiàng)目(No.81141097) 陜西省科技發(fā)展計劃項(xiàng)目(No.2012K13-01-09)~~
【分類號】:R739.63
[Abstract]:Objective to screen the proteomic differential expression of nasopharyngeal carcinoma cells with different radiosensitivity from the same source, and to explore the mechanism of radioresistance in nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell lines with different radiosensitivity were isolated and analyzed by two-dimensional gel electrophoresis and image analysis software PDQuest. Differential protein spots were identified by LC-MS/MS mass spectrometry and known differential proteins were classified by Gene Oncology Annotation. Some of the selected proteins were verified by Western blot. Results there were 27 candidate proteins in CNE-2 and CNE-2R cells, 11 of which were down-regulated and 16 up-regulated in radiation-resistant CNE-2R cells. Four of them: 4-3-3 蟽, GADD45,PRKDC and HSP27 were confirmed by Western blot method. Conclusion Radio-resistant cells were produced by repeated irradiation of CNE-2 cells, and some mutations occurred at the proteomic level, and these mutations were inherited stably, thus showing the phenotype of radioresistance. By regulating the expression of some proteins associated with radioresistance, it is possible to regulate the radiosensitivity of cells. 4-3-3 蟽, GADD45,PRKDC and HSP27 are potential gene markers for predicting the radiosensitivity of NPC. They may be involved in the production of radioresistance in NPC.
【作者單位】: 西安交通大學(xué)醫(yī)學(xué)院第二附屬醫(yī)院腫瘤病院放療科;
【基金】:國家自然科學(xué)基金資助項(xiàng)目(No.81141097) 陜西省科技發(fā)展計劃項(xiàng)目(No.2012K13-01-09)~~
【分類號】:R739.63
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2 范芳;安國順;李淑艷;倪菊華;賈弘y,
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