本文選題：葉酸　切入點：殼聚糖　出處：《廣西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究葉酸偶聯(lián)殼聚糖攜帶5′-氮雜-2′-脫氧胞苷(5-aza-2deoxycytidine,地西他濱,DAC)納米粒對比5′-氮雜-2′-脫氧胞苷作用于體外培養(yǎng)的人鱗狀上皮舌癌SCC-9細胞,了解葉酸偶聯(lián)殼聚糖載藥DAC對SCC-9細胞的p16基因和p16蛋白的表達情況,及對SCC-9細胞的抑制作用,從而探索葉酸偶聯(lián)殼聚糖載藥納米粒攜DAC對細胞的p16基因的高甲基化狀態(tài)的還原能力的有效性,以及同時了解載藥系統(tǒng)對細胞提高藥效的能力。方法:合成葉酸殼聚糖載藥DAC納米粒,測其zeta電位證實其為穩(wěn)定的納米顆粒后,將SCC-9細胞分為0、1、2、3、4、5、6、7八組。2、3、4組選用三個梯度濃度1×l0-7mol/L、1×l0-6mol/L、1×l0-5mol/L的DAC進行處理細胞,5、6、7組采用同樣三個濃度梯度1×l0-7mol/L、1×l0-6mol/L、1×l0-5mol/L的葉酸殼聚糖載藥DAC納米粒處理細胞,0組為空白生理鹽水對照組,1組為空白葉酸殼聚糖處理組,0組內(nèi)設(shè)3個濃度的組內(nèi)對照a、b、c,1組內(nèi)設(shè)3個組內(nèi)對照,為e、f、c組。觀察葉酸殼聚糖載藥DAC納米顆粒對比DAC作用細胞,用MTT法測各組藥物處理細胞5天內(nèi)的SCC-9細胞抑制率;藥物各自作用48小時后,實時熒光定量PCR法檢測細胞中p16基因m RNA的表達和應(yīng)用免疫組化SP法檢測細胞p16的蛋白表達。結(jié)果:MTT結(jié)果表明相同濃度的葉酸-殼聚糖載藥DAC納米顆粒組藥物對細胞的抑制率高于同樣濃度下的DAC組(P0.05),且藥物對細胞的抑制率均與藥物濃度呈正相關(guān)關(guān)系。實時熒光定量PCR結(jié)果顯示2、3、4、5、6、7組的p16基因m RNA表達量顯著高于0、1組(P0.05)。0組和1組無統(tǒng)計學(xué)意義(P0.05)。相同濃度的葉酸-殼聚糖載藥DAC納米顆粒組的p16基因的m RNA表達量高于同樣濃度下的DAC組(P0.05);免疫組化結(jié)果顯示,2、3、4、5、6、7組相較于0、1組呈強陽性表達,有統(tǒng)計學(xué)意義(P0.05),相同濃度的葉酸-殼聚糖載藥DAC納米顆粒組的p16蛋白的表達量高于同樣濃度下的DAC組并具有統(tǒng)計學(xué)意義(P0.05),0組和1組沒有統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1.葉酸-殼聚糖載藥納米粒攜帶DAC和DAC均對SCC-9細胞有明顯的去甲基化能力。2.葉酸-殼聚糖載藥納米粒攜帶DAC對比DAC具有更高效逆轉(zhuǎn)SCC-9細胞p16基因甲基化狀態(tài),并具有促使p16基因m RNA和蛋白恢復(fù)表達的作用。3.葉酸-殼聚糖載藥納米粒攜帶DAC和DAC,去甲基化能力沒有隨著藥物濃度升高而升高。4.葉酸-殼聚糖載藥納米粒攜帶DAC組對比DAC組,藥物抑制率更明顯。
[Abstract]:Aim: to study the effects of folic acid-coupled chitosan carrying 5-aza-2deoxycytidine (DAC-) nanoparticles on human squamous cell carcinoma (SCC-9) cells in vitro. To investigate the expression of p16 gene and p16 protein in SCC-9 cells and the inhibitory effect of folic acid-coupled chitosan loaded DAC on SCC-9 cells. To explore the effectiveness of folic acid-coupled chitosan loaded nanoparticles with DAC in reducing the hypermethylation of p16 gene. Methods: DAC nanoparticles loaded with chitosan folate were synthesized, and their zeta potential was determined to be stable nanoparticles. The SCC-9 cells were divided into 8 groups: 1 脳 l0-7 mol / L 1 脳 l0-6 mol / L 1 脳 1 脳 1 脳 L ~ (-1) mol / L ~ (1 脳 1 脳 10 ~ (-1) mol / L ~ (1)) l0-5mol/L. The cells were treated with the same concentration gradient of 1 脳 10 ~ (-7) mol / L ~ (-1) L ~ (-1) L _ (1 脳 10 ~ (-6) mol 路L ~ (-1)) DAC nanoparticles. Group 0 was treated as a blank normal saline control group (n = 1) with the same concentration gradient of 1 脳 10 ~ (-7) mol 路L ~ (-1) mol 路L ~ (-1) mol 路L ~ (-1) DAC. The control group was divided into 3 groups with 3 concentrations of folic acid chitosan treatment group, and 3 control groups with 3 concentrations in the control group (group 1), and the control group (group 1) was treated with folic acid chitosan. The DAC nanoparticles loaded with chitosan folate were observed to compare the effect of DAC on the cells. The inhibition rate of SCC-9 cells in each group was measured by MTT method within 5 days, and after 48 hours of treatment with each drug, the inhibition rate of SCC-9 cells was measured by MTT method. The expression of p16 gene m RNA was detected by real-time fluorescence quantitative PCR and the protein expression of p16 protein was detected by immunohistochemistry SP method. Results the results showed that the same concentration of folic acid-chitosan loaded DAC nanoparticles was used to detect the expression of p16 protein. The inhibition rate of cells was higher than that of DAC group at the same concentration, and the inhibitory rate of the drug on the cell was positively correlated with the drug concentration. The results of real-time quantitative PCR showed that the expression of p16 gene m RNA in group 2, 3, 3, 4, 5, 5, 5, 5, 7 was significantly higher than that in group 0, 0, 1, P0. 05 and 0. 0, respectively. The expression of m RNA of p16 gene in the same concentration of folic acid-chitosan loaded DAC nanoparticles was higher than that in the same concentration of DAC group P0.05.The immunohistochemical results showed that the expression of p16 gene was strongly positive in the same concentration of folate-chitosan loaded DAC group compared with the control group. The expression of p16 protein in the DAC nanoparticles with the same concentration of folic acid-chitosan was higher than that in the DAC group at the same concentration, and there was no significant difference in the expression of p16 protein between the two groups. Conclusion\\\%\\%\%\%\%\%\%? Both DAC and DAC carried by chitosan nanoparticles had obvious demethylation ability to SCC-9 cells. Compared with DAC, folic acid-chitosan nanoparticles carrying DAC could reverse the methylation of p16 gene in SCC-9 cells more efficiently. It can promote the expression of p16 gene m RNA and protein. 3. Folic acid-chitosan nanoparticles carry DAC and DAC, and the demethylation ability does not increase with the increase of drug concentration. 4. Folic acid-chitosan nanoparticles carrying DAC group compared with DAC group. The drug inhibition rate was more obvious.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.86

【參考文獻】

相關(guān)期刊論文 前10條

1 鄒文瑋;舒奇;傅穎媛;夏芝璐;;瑞香素毒理學(xué)實驗研究[J];中成藥;2013年05期

2 皮精英;陳超霞;彭強;彭大川;李宜靜;;子宮內(nèi)膜癌患者P16基因甲基化和血清CA125檢測的臨床意義[J];現(xiàn)代生物醫(yī)學(xué)進展;2011年18期

3 舒奇;傅穎媛;劉婷;余燕影;;金邊瑞香活性成分瑞香黃烷-Ⅰ,瑞香素對SMMC-7721和MCF-7細胞作用的研究[J];時珍國醫(yī)國藥;2009年09期

4 黃彬紅;傅穎媛;徐靜;曾小平;況南珍;熊麗紅;陳開森;周智興;劉婷;;金邊瑞香對荷S_(180)瘤小鼠免疫功能的影響[J];時珍國醫(yī)國藥;2008年10期

5 Koviljka Krtolica;Milena Krajnovic;Slavica Usaj-Knezevic;Dragan Babic;Dusan Jovanovic;Bogomir Dimitrijevic;;Comethylation of p16 and MGMT genes in colorectal carcinoma:Correlation with clinicopathological features and prognostic value[J];World Journal of Gastroenterology;2007年08期

6 王彩霞;馮霞;楊軍;;聚合物膠束型抗癌藥物給藥系統(tǒng)研究進展[J];化學(xué)研究與應(yīng)用;2006年06期

7 王銀松,李英霞,宋妮,張華;靶向抗腫瘤藥物甲氨喋呤-琥珀酰殼聚糖綴合物的制備及其體外穩(wěn)定性的初步探討[J];高等學(xué)�；瘜W(xué)學(xué)報;2003年11期

8 尹斌,沈巖;DNA的甲基化修飾與DNA甲基轉(zhuǎn)移酶[J];國外醫(yī)學(xué).遺傳學(xué)分冊;2001年01期

9 金哈斯,樓方定,于力;As_2O_3誘導(dǎo)急性淋巴細胞白血病細胞系Molt4細胞p15基因表達的研究[J];中華血液學(xué)雜志;2001年01期

10 白志勇,武淑蘭,徐國賓;白血病細胞DNA甲基轉(zhuǎn)移酶活性改變及其意義[J];中國實驗血液學(xué)雜志;1999年03期

，

本文編號：1665556