本文選題：卵巢癌　切入點(diǎn)：多藥耐藥　出處：《東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年02期

【摘要】：目的:構(gòu)建介孔二氧化硅(HMSN)復(fù)合載藥系統(tǒng),探究該系統(tǒng)對卵巢癌耐藥細(xì)胞株SKOV-3/ADR耐藥的逆轉(zhuǎn)作用。方法:在HMSN表面修飾羧基,通過機(jī)械攪拌和靜電吸引的作用包載ADR和熒光NVP;采用Zeta電位、透射電鏡等方法對此系統(tǒng)進(jìn)行表征,并測定此系統(tǒng)的包封率和載藥量以及在不同p H值環(huán)境中的藥物釋放率;再將實(shí)驗(yàn)分為兩組,即HMSN-COOH@ADR熒光NVP組和游離ADR熒光NVP組,采用熒光共聚焦觀察不同時(shí)間點(diǎn)HMSN復(fù)合載藥系統(tǒng)在胞內(nèi)的蓄積情況;利用MTT法檢測細(xì)胞在兩組作用24 h后的抑制率,同時(shí)通過流式細(xì)胞分析儀檢測兩組的凋亡率。結(jié)果:Zeta電位表明HMSN在修飾羧基之前其所帶電荷約為-20 mV,在修飾羧基后HMSN所帶電荷增加至約-40 mV;通過透射電鏡表明此載藥系統(tǒng)在包載藥物前后的形態(tài)和粒徑均未發(fā)生明顯改變;測定HMSN復(fù)合載藥系統(tǒng)中ADR的包封率為45%,載藥量為7.5%,熒光NVP的包封率為30%,載藥量為1.4%。HMSN復(fù)合載藥系統(tǒng)的藥物釋放率隨著環(huán)境p H值的降低而逐漸升高,具有顯著的p H敏感性,實(shí)現(xiàn)了以HMSN所處環(huán)境的p H值為"開關(guān)"調(diào)控藥物的釋放,有效地減少了復(fù)合載藥系統(tǒng)在胞外的非特異性釋放;通過熒光共聚焦觀察發(fā)現(xiàn)HMSN復(fù)合載藥系統(tǒng)作用細(xì)胞1 h后,其在胞內(nèi)的蓄積量即可明顯增加,隨著作用時(shí)間的延長胞內(nèi)的熒光強(qiáng)度也在逐漸增強(qiáng);藥物作用相同的時(shí)間,HMSN-COOH@ADR熒光NVP組較游離ADR熒光NVP組的MTT細(xì)胞抑制率和細(xì)胞凋亡率均明顯提高(P0.05),對細(xì)胞的毒性作用明顯增強(qiáng)。結(jié)論:構(gòu)建的HMSN-COOH@ADR熒光NVP的復(fù)合載藥系統(tǒng)可以有效逆轉(zhuǎn)卵巢癌耐藥細(xì)胞株SKOV-3/ADR的耐藥性,其逆轉(zhuǎn)耐藥的機(jī)制是通過非特異性的內(nèi)吞途徑攜帶復(fù)合載藥系統(tǒng)進(jìn)入細(xì)胞內(nèi),逃避了P-gp等ABC運(yùn)載蛋白超家族的識(shí)別和外排作用。
[Abstract]:Aim: to construct a mesoporous silica HMSN complex drug carrier system and to investigate the reversal effect of the system on drug resistance of ovarian cancer cell line SKOV-3/ADR.Methods: the carboxyl group was modified on the surface of HMSN. The ADR and fluorescence were encapsulated by mechanical agitation and electrostatic attraction. The system was characterized by Zeta potential and transmission electron microscopy.The encapsulation efficiency, drug loading capacity and drug release rate in different pH values were determined, and then divided into two groups: HMSN-COOH@ADR fluorescent NVP group and free ADR fluorescent NVP group.The intracellular accumulation of HMSN compound drug loading system at different time points was observed by fluorescence confocal method, and the inhibition rate of cells in the two groups after 24 h exposure was detected by MTT assay, and the apoptosis rate of the two groups was detected by flow cytometry.The entrapment efficiency of ADR was 455.The entrapment efficiency of fluorescent NVP was 30. The drug release rate of 1.4%.HMSN compound drug loading system increased with the decrease of pH value and had significant sensitivity to pH.The release of the drug was regulated by the pH value of the environment in which HMSN was located, and the non-specific release of the compound drug loading system was effectively reduced, and it was found that the HMSN compound drug carrier system acted on the cells for 1 hour after fluorescence confocal observation.The amount of intracellular fluorescence increased obviously, and the fluorescence intensity increased gradually with the prolongation of the action time.The inhibition rate and apoptosis rate of MTT cells in HMSN-COOHADR fluorescent NVP group were significantly higher than those in free ADR fluorescent NVP group, and the cytotoxic effect of HMSN-COOHADR fluorescent NVP group was obviously enhanced.Conclusion: the complex drug loading system of HMSN-COOH@ADR fluorescent NVP can effectively reverse the drug resistance of ovarian cancer cell line SKOV-3/ADR. The mechanism of reversing drug resistance is to carry the complex drug carrier system into the cell through non-specific endocytosis pathway.It evades the recognition and efflux of ABC transporter superfamily such as P-gp.
【作者單位】： 東南大學(xué)醫(yī)學(xué)院;東南大學(xué)附屬中大醫(yī)院婦產(chǎn)科;
【分類號(hào)】：R737.31
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本文編號(hào)：1697347