本文選題：羥基磷灰石 + P53　； 參考：《中南大學(xué)》2010年碩士論文

【摘要】： 目的：探討經(jīng)聚乙烯亞胺修飾的納米羥基磷灰石-P53復(fù)合物轉(zhuǎn)染鼻咽癌細胞(HNE2系)的效果。 方法：(1)構(gòu)制pEGFP-C1-P53質(zhì)粒；(2)用pEGFP-C1-P53質(zhì)粒和經(jīng)聚乙烯亞胺修飾的納米羥基磷灰石(簡稱：PEI-nHA)混合制成羥基磷灰石-P53復(fù)合物,用于鼻咽癌HNE2系細胞的轉(zhuǎn)染；然后檢測轉(zhuǎn)染后的鼻咽癌HNE2細胞中P53轉(zhuǎn)染效率。(3) RT-PCR和Western-blot實驗檢測納米羥基磷灰石和脂質(zhì)體2000介導(dǎo)P53基因在基因和蛋白質(zhì)水平的表達。 結(jié)果：(1)在轉(zhuǎn)染實驗中轉(zhuǎn)染后的空白組(單純質(zhì)粒和HNE2細胞混合)在熒光顯微鏡下檢測未見熒光,脂質(zhì)體2000組在熒光顯微鏡下檢測可見40%左右細胞內(nèi)具有綠色熒光,納米載體組分成62.5ug/ml、125ug/ml、250ug/ml、500ug/ml四種濃度轉(zhuǎn)染,在熒光顯微鏡下檢測經(jīng)四種不同濃度納米材料介導(dǎo)轉(zhuǎn)染HNE2細胞后,62.5ug/ml濃度的PEI-nHA轉(zhuǎn)染效率約為4%,125 ug/ml濃度的PEI-nHA轉(zhuǎn)染效率約為6%,250ug/ml濃度的PEI-nHA轉(zhuǎn)染效率約為5%,500 ug/ml濃度的PEI-nHA轉(zhuǎn)染效率約為1%。(2) RT-PCR和Western-blot實驗經(jīng)脂質(zhì)體2000、納米羥基磷灰石介導(dǎo)轉(zhuǎn)染進鼻咽癌HNE2細胞均可檢測到P53在基因和蛋白水平的表達,經(jīng)脂質(zhì)體2000介導(dǎo)轉(zhuǎn)染的HNE2細胞的P53基因和蛋白表達水平較納米羥基磷灰石載體介導(dǎo)的明顯增強。 結(jié)論：(1)經(jīng)聚乙烯亞胺修飾的納米羥基磷灰石介導(dǎo)轉(zhuǎn)染鼻咽癌HNE2細胞的轉(zhuǎn)染率較脂質(zhì)體2000偏低,有待進一步研究。
[Abstract]:Aim: to investigate the effect of poly (imine) modified nano-hydroxyapatite-p53 complex transfected into nasopharyngeal carcinoma cell line (HNE2). Methods the pEGFP-C1-P53 plasmids were constructed by 1) the pEGFP-C1-P53 plasmids were mixed with polyethylene imide modified nano-hydroxyapatite to form hydroxyapatite p53 complexes, which were used for transfection of nasopharyngeal carcinoma (NPC) HNE2 cells. Then the transfection efficiency of p53 in nasopharyngeal carcinoma (HNE2) cells was detected by RT-PCR and Western-blot. The expression of p53 gene mediated by nano-hydroxyapatite and liposome 2000 was detected at the gene and protein levels. Results in the transfection experiment, no fluorescence was detected in the blank group (simple plasmids and HNE2 cells mixed), while in the liposome 2000 group, about 40% of the cells showed green fluorescence under the fluorescence microscope. The nanocarrier group was transfected into four concentrations of 62.5ug / ml 125ugrml 250ugpml / ml 500ugrml, After transfection of HNE2 cells with four different concentrations of nanomaterials, the transfection efficiency of PEI-nHA was about 42.5 渭 g / ml, and the transfection efficiency of PEI-nHA was about 62.5 ug/ml / ml. The transfection efficiency of PEI-nHA at 250ug-1 / ml concentration was about 5500 ug/ml / ml PEI-nHA transfection efficiency. The efficiency of RT-PCR and Western-blot experiments was about 1: 1. RT-PCR and Western-blot were transfected into HNE2 cells of nasopharyngeal carcinoma by liposome 2000. the expression of p53 at gene and protein level could be detected by nano-hydroxyapatite transfection into nasopharyngeal carcinoma HNE2 cells. The expression level of p53 gene and protein in HNE2 cells transfected with liposome 2000 was significantly higher than that mediated by nano-hydroxyapatite vector. Conclusion the transfection efficiency of HNE2 cells mediated by polyethylene imine modified nano-hydroxyapatite is lower than that of liposome 2000, which needs further study.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R739.63

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