發(fā)布時(shí)間：2018-06-07 03:15

  本文選題：人類表皮生長(zhǎng)因子2陽(yáng)性 + 胃癌��； 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：背景： 胃癌是嚴(yán)重危害人類生命健康的重大疾病,世界衛(wèi)生組織癌控項(xiàng)目數(shù)據(jù)顯示,全球每年胃癌新發(fā)病例約93萬(wàn),死亡人數(shù)約70萬(wàn),占所有癌癥死亡率的1/10;我國(guó)每年新發(fā)胃癌患者達(dá)40萬(wàn)人,死亡人數(shù)達(dá)30萬(wàn),患病率和死亡率均是世界平均水平的2倍多。目前在我國(guó)胃癌早期檢出率不到10%。超過(guò)90%以上的胃癌患者于就診時(shí)已是進(jìn)展期以上,或在確診時(shí)已經(jīng)發(fā)生了轉(zhuǎn)移或播散,從而失去了手術(shù)的機(jī)會(huì)。因此,胃癌的化學(xué)治療是國(guó)內(nèi)絕大多數(shù)胃癌患者的主要治療方法,在我國(guó)有著重要的研究意義。但是,當(dāng)前臨床應(yīng)用的常規(guī)化療藥物全身毒副作用較大,患者耐受性差；腫瘤細(xì)胞對(duì)化療藥物產(chǎn)生耐藥等,都容易導(dǎo)致化療的失敗。針對(duì)胃癌常規(guī)化療藥物的全身用量大,毒副作用大的研究,具有極大地學(xué)術(shù)價(jià)值、社會(huì)效應(yīng)及應(yīng)用前景。近年來(lái),由于各種納米粒子能理想地在體內(nèi)傳遞各種物質(zhì)并具有可調(diào)的表面化學(xué)靶向結(jié)構(gòu),使得它在腫瘤治療中凸顯出獨(dú)特的優(yōu)勢(shì)。 目的： 本研究在胃癌細(xì)胞人類表皮生長(zhǎng)因子2受體為靶向的基礎(chǔ)上,以貴金屬Au納米顆粒通過(guò)交換偶合與Fe304形成納米復(fù)合物作為載體,化學(xué)結(jié)合單克隆抗體赫賽汀及一線化療藥物奧沙利鉑,從而構(gòu)建以人類表皮生長(zhǎng)因子2為靶向的磁共振成像示蹤控釋奧沙利鉑的新型納米復(fù)合物Oxaliplatin-Au-Fe3O4-Herceptin,并且評(píng)價(jià)其在人類表皮生長(zhǎng)因子2陽(yáng)性胃癌中的價(jià)值。 方法： 1.應(yīng)用交換偶合法及熱降解技術(shù)在金表面降解乙酰丙酮鐵完成Au-Fe3O4金屬納米復(fù)合物的構(gòu)建。通過(guò)表面修飾劑的加入對(duì)Au-Fe3O4納米顆粒的形貌進(jìn)行調(diào)控。透射電鏡表征納米粒子。 2.液相Oxaliplatin-Au-Fe3O4-Herceptin與1-(3-二甲氨基丙基)-3-乙基二亞胺及N-羥基硫代琥珀�；旌闲纬蛇B接鍵,再與赫賽汀溶液混合培養(yǎng),形成Au-Fe3O4-Herceptin�？焖賹游龇兓玫綂W沙利鉑連接配體�；旌蠆W沙利鉑混懸液、Au-Fe3O4-Herceptin及連接配體,優(yōu)化比例,培養(yǎng)純化得到Oxaliplatin-Au-Fe3O4-Herceptin。MALDI-MS驗(yàn)證產(chǎn)物,體外透析袋不同pH值下檢測(cè)奧沙利鉑結(jié)合效率。 3. MTS法分別驗(yàn)證Au-Fe3O4、Au-Fe3O4-Herceptin Oxaliplatin-Au-Fe3O4-Herceptin細(xì)胞毒性。 4.常規(guī)培養(yǎng)HER2陽(yáng)性胃癌細(xì)胞系SGC-7901。分別與Au-Fe3O4、 Oxaliplatin-Au-Fe3O4-Herceptin孵育2小時(shí),透射電鏡觀察納米復(fù)合物在胃癌細(xì)胞表面胞吞作用的程度。 5.胃癌原位移植瘤模型的建立及分組：研究不同時(shí)限注射Oxaliplatin-Au-Fe3O4-Herceptin的胃癌裸鼠模型,調(diào)整建立磁共振掃描參數(shù),觀察小鼠腫瘤組織變化。結(jié)果 1.透射電鏡下,見(jiàn)納米粒子大小約25nm,鐵粒子與金粒子1:1恒定； 2.成功構(gòu)建了Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物,并通過(guò)質(zhì)譜、紅外及透析袋緩釋奧沙利鉑證實(shí)了各分子間通過(guò)化學(xué)連接鍵相連。細(xì)胞毒性實(shí)驗(yàn)顯示：當(dāng)鐵離子濃度在30ug/ml以內(nèi)時(shí),Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物對(duì)細(xì)胞基本沒(méi)有毒性； 3.體外胞吞實(shí)驗(yàn)：將Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物、Au-Fe3O4納米粒子以相同鐵粒子濃度與SGC-7901細(xì)胞共孵育2h后透射電鏡可見(jiàn),Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物被大量吞入細(xì)胞,而Au-Fe3O4納米粒子未見(jiàn)被吞噬入細(xì)胞內(nèi)； 4.小鼠尾靜脈注射納米粒子復(fù)合物一段時(shí)間后,磁共振掃描老鼠皮下移植瘤,見(jiàn)腫瘤組織內(nèi)T2相低密度信號(hào)灶。腫瘤組織普魯士染色后可見(jiàn)腫瘤血管旁大量鐵粒子聚集。結(jié)論： 本課題通過(guò)學(xué)科交叉研究,研制成功針對(duì)人類表皮生長(zhǎng)因子2受體金屬納米化合物Oxaliplatin-Au-Fe3O4-Herceptin,能結(jié)合奧沙利鉑靶向胃癌,并實(shí)現(xiàn)磁共振示蹤控釋,獲得擁有自主知識(shí)產(chǎn)權(quán)。研究明確了該種化合物的體內(nèi)外作用作用機(jī)制和生物安全性,為進(jìn)一步的臨床應(yīng)用和人類表皮生長(zhǎng)因子2陽(yáng)性胃癌患者的個(gè)性化治療,以及新藥的開發(fā)提供堅(jiān)實(shí)的實(shí)驗(yàn)基礎(chǔ)。同時(shí),亦為開發(fā)新型靶向治療藥物和靶向治療提供新的思路和技術(shù)支持。
[Abstract]:Background :

Gastric cancer is a major disease which seriously endangers the health of human life . The data of World Health Organization Cancer Control Project shows that the number of new cases of gastric cancer in the world is about 9.3 million , the number of deaths is about 700,000 , accounting for 1 / 10 of all cancer mortality . In our country , more than 90 % of patients with gastric cancer have progressed more than 10 % each year .
In recent years , various kinds of nano - particles can transfer various substances in vivo and have adjustable surface chemical targeting structure , so that it has unique advantages in tumor treatment .

Purpose :

Based on the targeting of human epidermal growth factor - 2 receptor in gastric cancer cells , a novel nano - complex Oxhide - Au - Fe _ 3O _ ( 4 - -Fe _ 3O _ 3O _ 4 - ) was prepared by exchange coupling with Fe304 as a carrier , and it was used as a carrier to construct a novel nano - composite Oxhide - Au - Fe3O4 - Hercee with magnetic resonance imaging and release oxaliplatin targeting human epidermal growth factor 2 , and its value in human epidermal growth factor 2 positive gastric cancer was evaluated .

Method :

1 . The Au - Fe _ 3O _ 4 metal nanoparticles were prepared by using exchange coupling method and thermal degradation technique on gold surface . The morphology of Au - Fe3O4 nanoparticles was controlled by the addition of surface modifier . The nano - particles were characterized by TEM .

2 . The preparation of Oxhide - Au - Fe _ 3O _ 3 - ethyldiimine with 1 - ( 3 - dimethylaminopropyl ) -3 - ethyldiimine and N - hydroxythiosuccinyl was carried out to form the bond , and then mixed with Hercystatin solution to form Au - Fe _ 3O _ 3O _ 4 - HerceII . Rapid chromatographic method was used to purify the oxaliplatin - linked ligand .

3 . The toxicity of Au - Fe _ 3O _ 4 , Au - Fe _ 3O _ 3O _ 3O _ 3 - 2 - Fe _ 3O _ 3O _ 3O _ 3 - 4 - Fe _ 3O _ 3O _ 4 - 2 was verified by MTS method .

4 . The HER2 - positive gastric cancer cell line SGC - 790was incubated with Au - Fe _ 3O _ 3O _ 4 and Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 3O _ 4 for 2 hours , and the extent of endocytosis of the nano - complexes on the surface of gastric cancer cells was observed by transmission electron microscope .

5 . Establishment and grouping of in - situ transplanted tumor model of gastric cancer : The nude mouse model of gastric cancer with different time - frame injection of Oxhide - Au - Fe3O4 - HerceII was studied , and the parameters of magnetic resonance scan were adjusted to observe the changes of tumor tissues in mice .

1 . Under the transmission electron microscope , the size of the nanoparticles is about 25nm , and the iron particles are constant with the gold particles 1 : 1 ;


2 . Oxhide - Au - Fe _ 3O _ 4 - Hertin nanocomposites were successfully constructed . Through mass spectrometry , IR and slow release of oxaliplatin , it was confirmed that there was no toxicity to the cells .


3 . In vitro endocytosis experiment : Oxhide - Au - Fe _ 3O _ 3O _ 4 - Hertin nanocomposites were incubated for 2 hours at the same concentration of Fe particles and SGC - 790cells for 2 hours . The Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 4 nanoparticles were absorbed into the cells and the Au - Fe _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 4 nanoparticles were swallowed into the cells .


4 . After intravenous injection of nano - particle complex for a period of time , magnetic resonance scanning the mouse subcutaneous transplantation tumor to see the T2 - phase low - density signal focus in the tumor tissue . After the tumor tissue , the tumor tissue was stained and the large amount of iron particles near the tumor vessel were collected . Conclusion :

The study on the mechanism and biological safety of human epidermal growth factor - 2 receptor metal nano - compound Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 4 - Her2 combined with oxaliplatin targeted gastric cancer , and achieved independent intellectual property rights . It also provides a solid experimental basis for further clinical application and personalized treatment of human epidermal growth factor 2 positive gastric cancer patients , and provides new ideas and technical support for the development of new targeted therapy drugs and targeted therapy .
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：TB383.1

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 陽(yáng)諾;冼磊;陳銘伍;郭建極;王令平;徐輝航;;非小細(xì)胞肺癌患者Bcl-2蛋白的表達(dá)及意義[J];廣西醫(yī)學(xué);2011年05期

2 劉雄雄;李強(qiáng);;氧化應(yīng)激引起的遺傳變化和DNA甲基化改變?cè)谀[瘤發(fā)生中的作用[J];輻射研究與輻射工藝學(xué)報(bào);2013年06期

3 張亮;杜佳;周立為;程q，

本文編號(hào)：1989494