北京生物醫(yī)學(xué)工程

多肽修飾的金納米顆粒對(duì)小鼠三陰性乳腺癌細(xì)胞的靶向光熱效應(yīng)研究

Peptide modified gold nanoparticles enhancethe elimination of triple-negative breast cancercellsthrough photothermal effect

作者：孫博蒙藝靈溫濤孟潔劉健許海燕

單位：中國(guó)醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所，北京協(xié)和醫(yī)學(xué)院基礎(chǔ)學(xué)院（北京 100005） 通信作者：許海燕，E-mail：[email protected]；溫濤，E-mail：[email protected]

關(guān)鍵詞：熱休克蛋白60；乳腺癌細(xì)胞；光熱；靶向殺傷

分類號(hào)：R318.04

出版年·卷·期（頁(yè)碼）：2021·40·5（441-446）

摘要：

目的乳腺癌細(xì)胞高表達(dá)熱休克蛋白60（HSP60）。本文用可特異性結(jié)合的多肽P17修飾金納米顆粒（AuNP），構(gòu)建靶向光熱治療系統(tǒng)（AuNP-P17），研究其對(duì)小鼠乳腺癌細(xì)胞4T1的光熱殺傷作用。方法利用紫外-可見(jiàn)-近紅外分光光度計(jì)、動(dòng)態(tài)光散射儀（dynamic light scattering，DLS）和透射電子顯微鏡（transmission electron microscope，TEM）對(duì)納米顆粒的光學(xué)性質(zhì)、粒徑和電勢(shì)進(jìn)行表征；通過(guò)熒光標(biāo)記和流式細(xì)胞術(shù)檢測(cè)細(xì)胞HSP60表達(dá)水平；利用光學(xué)顯微鏡觀察4T1細(xì)胞對(duì)納米顆粒的攝取；使用便攜式熱電偶溫度計(jì)測(cè)量細(xì)胞所在溶液在激光照射下的升溫曲線；采用細(xì)胞增殖-毒性檢測(cè)法（cell counting Kit-8，CCK-8）測(cè)定細(xì)胞活性。結(jié)果金納米顆粒呈均勻球形，水合粒徑為（51.8 ± 0.1）nm；表面連接P17后（AuNP-P17）水合粒徑為（52.1 ± 2.2）nm。在金原子濃度不大于100 μg/mL范圍內(nèi)，AuNP和AuNP-P17與4T1細(xì)胞共孵育24 h對(duì)細(xì)胞活性無(wú)顯著性影響。相比于AuNP，AuNP-P17被更多地?cái)z取到細(xì)胞中。激光照射4 min后，AuNP-P17、AuNP組和對(duì)照組的溶液溫度從（22.9 ± 0.4）°C分別上升到（52.0 ± 3.9）°C、（47.2 ± 0.6）°C和（38.2 ± 2.4）°C，4T1細(xì)胞存活率分別為（1.3 ± 2.8）%、（64.1 ± 6.8）%和（100.0 ± 3.2）%，兩兩比較差異具有統(tǒng)計(jì)學(xué)意義（P<0.01）。

結(jié)論 AuNP-P17可被4T1細(xì)胞攝取，具有良好的生物相容性、靶向性和光熱效應(yīng)。在激光照射下，對(duì)乳腺癌細(xì)胞具有更強(qiáng)的殺傷作用，在乳腺癌光熱治療方面具有轉(zhuǎn)化應(yīng)用潛力。

Objective Breast cancer cells highly express heat shock protein 60 (HSP60). In this paper, we construct a targeted photothermal therapy system by modifying gold nanoparticle (AuNP) with a HSP60 specific binding peptide (P17) and study the photothermal killing effect on breast cancer cell (4T1).Methods The optical properties, particle size and potential of nanoparticles were characterized by UV-Vis-NIR spectrophotometer, dynamic light scattering (DLS) and transmission electron microscope (TEM). The expression of HSP60 was detected by fluorescence labeling and flow cytometry. The uptake of nanoparticles by 4T1 cells was observed by light microscope. The heating curve of cell solution under laser irradiation was measured by portable thermocouple thermometer. The cell viability was detected by cell counting kit-8 (CCK-8).Results The gold nanoparticles are spherical and the hydrated particle size is (51.8 ± 0.1) nm. The hydrated particle size of AuNP-P17 is (52.1 ± 2.2) nm. After incubated with AuNP and AuNP-P17 for 24h, the cellular viability of 4T1 had no significant change when the concentration of gold atom was lower than 100 μg/mL. The uptake of AuNP-P17 by 4T1 cells was observed much higher than that of AuNP with light microscope. With laser irradiation for 4 minutes, the temperature of 4T1 cells incubated with AuNP-P17, AuNP and the control group increased from (22.9 ± 0.4) °C to (52.0 ± 3.9) °C, (47.2 ± 0.6) °C and (38.2 ± 2.4) °C, respectively. The survival rate of 4T1 cells changed to (1.3 ± 2.8) %, (64.1 ± 6.8) %, and (100.0 ± 3.2) % with significant difference (P < 0.01).Conclusions AuNP-P17 can be uptake by 4T1, and has good biocompatibility, targeting and photothermal effect. With laser irradiation, it can promote the targeted killing effect of AuNP-P17 on 4T1 cells, which has the potential of transformation and application in the treatment of photothermal of breast cancer.

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