红色荧光碳量子点用于肿瘤微酸环境诊断

doi:10.3866/PKU.WHXB201905067

摘要/Abstract

摘要：

实现肿瘤的早期诊断对于提高癌症患者的存活率或延长生存时间非常重要。由于肿瘤细胞的糖酵解失调，导致肿瘤组织呈现微酸环境(pH < 6.8)，因此可以通过设计在pH 6.8响应的荧光材料实现对于肿瘤的早期诊断。在本工作中，我们以4-二甲氨基苯酚为前驱体，高碘酸钾为氧化剂，采用溶剂热法制备了发射波长为640 nm的水溶性红色荧光碳量子点(R-CQDs)。将单甲醚聚乙二醇衍生物(MeO-PEG-PDPA)修饰到R-CQDs表面，获得在pH 6.8响应的R-CQDs (pRF-R-CQDs)，pH < 6.8时呈现红色荧光，pH > 6.8时荧光淬灭。通过透射电子显微镜(TEM)、X射线光电子能谱(XPS)、傅里叶变换红外(FT-IR)光谱、拉曼(Raman)光谱、X射线粉末衍射(XRD)图谱、荧光(PL)光谱、紫外可见(UV-Vis)吸收光谱对R–CQDs进行结构与性质测定。研究了pRF-R-CQDs在组织成像及活体成像中的应用。结果表明：肿瘤微酸环境(pH < 6.8)中，pRF-R-CQDs的氨基质子化而呈现红色荧光。正常组织(pH > 6.8)中，pRF-R-CQDs的氨基去质子化导致荧光淬灭。pRF-R-CQDs的浓度在高达100 μg·mL^-1时仍未表现出明显的细胞毒性。pRF-R-CQDs能够用于区分出肿瘤和正常组织，可用于肿瘤的早期诊断。

关键词: 红色荧光, 碳量子点, pH响应, 微酸环境, 肿瘤早期诊断

Abstract:

Cancer remains a major global cause of morbidity and mortality. Diagnosis at an early stage can significantly improve the survival of cancer patients. Cancers of different origins often have vastly different genotypes and phenotypes. Therefore, it is challenging to establish a universal strategy for cancer detection. Universal cancer detection can be potentially achieved by using pH-responsive probes. An acidic microenvironment is mainly caused by lactic acid accumulation in rapidly growing tumor cells. Based on the difference in pH between tumor and normal tissues, fluorescent materials that respond to a pH of around 6.8 are ideal for tumor detection. Carbon quantum dots (CQDs) have attracted much attention in bioimaging owing to their outstanding characteristics such as stable photoluminescence, low cytotoxicity, excellent biocompatibility, and resistance to photobleaching. In this study, red fluorescent CQDs (R-CQDs) were synthesized by the solvothermal treatment of 4-(dimethylamino) phenol in the presence of potassium periodate. The UV-Vis spectrum of the R-CQDs showed a characteristic absorption peak at 545 nm. The photoluminescence spectrum revealed an emission peak at 640 nm. The brightness of this photoluminescence peak was quantified to be 12.8% in terms of the absolute quantum yield (QY). Transmission electron microscopy (TEM) images showed that the R-CQDs have uniform sizes with an average diameter of 4 nm and a lattice spacing of 0.21 nm. Fourier transform infrared (FT-IR) spectroscopy and X-ray photoelectron spectroscopy (XPS) confirmed that the R-CQDs have a large number of carboxyl groups. The Raman spectrum of the R-CQDs showed the characteristic D band at 1340 cm^-1 and G band at 1585 cm^-1. The X-ray powder diffraction (XRD) pattern showed a broad (002) peak centered at around 23°. The R-CQDs were responsive to highly acidic or alkaline conditions. The incorporation of a block copolymer (MeO-PEG-PDPA), prepared by atom transfer radical polymerization (ATRP), on the R-CQDs produced pH-responsive fluorescent CQDs (pRF-R-CQDs). Photoluminescence (PL) spectra showed that the pRF-R-CQDs were responsive at pH 6.8. At pH > 6.8, the fluorescence of the pRF-R-CQDs would be quenched because of deprotonation of the amine groups. In contrast, protonation of the amine groups would lead to a dramatic increase in fluorescence emission. TEM images showed that the pRF-R-CQDs self-assemble and disassemble at pH 6.8 because of their pH-responsive properties. Compared with most existing fluorescent materials, the pRF-R-CQDs can effectively resist photobleaching and autofluorescence. Moreover, these pRF-R-CQDs have minimal toxicity and can distinguish tumors from normal tissues. Therefore, pRF-R-CQDs have great potential for use as a universal material in tumor microenvironment diagnosis.

Key words: Red fluorescence, Carbon quantum dots, pH-response, Acidic microenvironment, Early diagnosis of Tumor

黄靖, 王丹阳, 李淑花, 范红, 范楼珍. 红色荧光碳量子点用于肿瘤微酸环境诊断[J]. 物理化学学报, 2021, 37(10), 1905067. doi: 10.3866/PKU.WHXB201905067

Jing Huang, Danyang Wang, Shuhua Li, Hong Fan, Louzhen Fan. Red Fluorescent Carbon Quantum Dots for Diagnosis of Acidic Microenvironment in Tumors[J]. Acta Phys. -Chim. Sin. 2021, 37(10), 1905067. doi: 10.3866/PKU.WHXB201905067

链接本文: https://www.whxb.pku.edu.cn/CN/10.3866/PKU.WHXB201905067

https://www.whxb.pku.edu.cn/CN/Y2021/V37/I10/1905067

图/表 6

图1

图2

图3

图4

图5

图6

参考文献 46

1	Hirsch F. R. ; Franklin W. A. ; Gazdar A. F. ; Bunn P. A Clin. Cancer Res 2001, 7, 5.
2	Nothacker M. ; Duda V. ; Hahn M. ; Warm M. ; Degenhardt F. ; Madjar H. ; Weinbrenner S Cancer 2009, 9, 335. doi: 10.1186/1471-2407-9-335
3	Nguyen Q. T. ; Tsien R. Y Nat. Rev. Cancer 2013, 13, 653. doi: 10.1038/nrc3566
4	Shariat S. F. ; Karakiewicz P. I. ; Ashfaq R. ; Lerner S. P. ; Palapattu G. S. ; Cote R. J. ; Sagalowsky A. I. ; Lotan Y Cancer 2008, 112, 315. doi: 10.1002/cncr.23162
5	Cheang M. C. U. ; Voduc D. ; Bajdik C. ; Leung S. ; McKinney S. ; Chia S. K. ; Perou C. M. ; Nielsen T. O Clin. Cancer Res 2008, 14, 1368. doi: 10.1158/1078-0432.CCR-07-1658
6	Li T. F. ; Li Y. W. ; Xiao L. ; Yu H. T. ; Fan L. Z Acta Chim. Sin 2014, 72, 227. doi: 10.6023/A13101036
7	Peppercorn J. ; Shapira I. ; Deshields T. ; Kroetz D. ; Friedman P. ; Spears P. ; Collyar D. ; Shulman L. ; Dressler L. ; Bertagnolli M Cancer 2012, 118, 5060. doi: 10.1002/cncr.27515
8	Arya S. K. ; Bhansali S Chem Rev. 2011, 111, 6783. doi: 10.1021/cr100420s
9	Stephan C. ; Cammann H. ; Meyer H. A. ; Lein M. ; Jung K Cancer Lett 2007, 249, 18. doi: 10.1016/j.canlet.2006.12.031
10	Shukla H. D. ; Mahmood J. ; Vujaskovic Z Cancer Lett 2015, 369, 28. doi: 10.1016/j.canlet.2015.08.003
11	He P. ; Yuan F. L. ; Wang Z. F. ; Tan Z. A. ; Fan L.Z. Acta Phys. -Chim. Sin 2018, 34 (11), 1250.
	贺平; 袁方龙; 王子飞; 谭占鳌; 范楼珍. 物理化学学报, 2018, 34 (11), 1250. doi: 10.3866/PKU.WHXB201804041
12	Vaupel, P.; Kallinowski, F.; Okunieff, P. Cancer Res. 1989, 49, 6449. doi: content/49/23/6449
13	Denko N. C Nat. Rev. Cancer 2008, 8, 705. doi: 10.1038/nrc2468
14	Bissell M. J. ; Hines W. C Nat. Med 2011, 17, 320. doi: 10.1038/nm.2328
15	Wang L. ; Li C J. Mater. Chem 2011, 21, 15862. doi: 10.1039/c1jm12072g
16	Webb B. A. ; Chimenti M. ; Jacobson M. P. ; Barber D. L Nat. Rev. Cancer 2011, 11, 671. doi: 10.1038/nrc3110
17	Vander Heiden M. G. ; Cantley L. C. ; Thompson C. B Science 2009, 324, 1029. doi: 10.1126/science.1160809
18	Kroemer G. ; Jaattela M Nat. Rev. Cancer 2005, 5, 886.
19	Tannock I. F. ; Rotin D Cancer Res 1989, 49, 4373.
20	Zhang X. ; Lin Y. ; Gillies R. J J. Nucl. Med 2010, 51, 1167. doi: 10.2967/jnumed.109.068981
21	Montet X. ; Ntziachristos V. ; Grimm J. ; Weissleder R Cancer Res 2005, 65, 6330. doi: 10.1158/0008-5472
22	Svoronos A. ; Braddock D. T. ; Glazer P. M. ; Engelman D. M. ; Saltzman W. M. ; Slack F. J Nature 2015, 518, 107. doi: 10.1038/nature13905
23	Xie W. J. ; Fu Y. Y. ; Ma H. ; Zhang M. ; Fan L. Z Acta Chim. Sin 2012, 70, 2169. doi: 10.6023/A12060302
24	Yuan Y. ; Ding D. ; Li K. ; Liu J. ; Liu B Small 2014, 10, 1967. doi: 10.1002/smll.201302765
25	Urano Y. ; Asanuma D. ; Hama Y. ; Koyama. Y ; Kamiya. M. ; Nagano T. ; Hasegawa A Nat. Med 2009, 15, 104. doi: 10.1038/nm.1854
26	Reshetnyak Y. K. ; Andreev O. A. ; Lehnert U. ; Engelman D. M Proc. Nat. Acad. Sci. U.S.A. 2006, 103, 6460. doi: 10.1073/pnas.0601463103
27	Zhang M. ; Bai L. ; Shang W. ; Xie W ; Fang D. ; Sun H. ; Fan L. ; Han M. ; Liu C. ; Yang S J. Mater. Chem 2012, 22, 7461. doi: 10.1039/c2jm16835a
28	Fan Z. ; Li Y. ; Li X. ; Fan L. ; Zhou S. ; Fang D. ; Yang S Carbon 2014, 70, 149- 156. doi: 10.1016/j.carbon.2013.12.085
29	Yuan F. ; Wang Z. ; Li S. ; Tan Z. ; Fan L. ; Yang S Adv. Mater 2017, 29, 1604436. doi: 10.1002/adma.201604436
30	Yuan F. ; Yuan T. ; Sui L. ; Wang Z. ; Fan L. ; Tan Z Nat. Commun. 2018, 9, 2249. doi: 10.1038/s41467-018-04635-5
31	Gong N. ; Ma X. ; Ye X. ; Zhou Q. ; Chen X. ; Tan X. ; Yao S. ; Huo S ; Zhang T. ; Chen S. ; et al Nat. Nanotech 2019, 14, 379. doi: 10.1038/s41565-019-0373-6
32	Chen D. ; Feng H. B. ; Li J Chem. Rev 2012, 112, 6027. doi: 10.1021/cr300115g
33	Xi Z. F. ; Yuan F. L. ; Wang Z. F ; Li S.H. ; Fan L. Z Acta Chim. Sin 2018, 76, 460. doi: 10.6023/A18020048
34	Fan Z. ; Zhou S. ; Garcia C. ; Fan L. ; Zhou J Nanoscale 2017, 9, 4928. doi: 10.1039/c7nr00888k
35	Li S. ; Zhou S. ; Li Y. ; Zhu J. ; Fan L. ; Yang S ACS Appl. Mater. Interfaces 2017, 9, 22332. doi: 10.1021/acsami.7b07267
36	Sun M. ; Liu H. ; Liu Y. ; Qu J. ; Li J Nanoscale 2015, 7, 1250. doi: 10.1039/c4nr05838k
37	Helmlinger G. ; Yuan F. ; Dellian M. ; Jain R. K Nat. Med 1997, 3, 177. doi: 10.1038/nm0297-177
38	Volk T. ; Jahde E. ; Fortmeyer H. P. ; Glusenkamp K. H. ; Rajewsky M. F Br. J. Cancer 1993, 68, 492. doi: 10.1038/bjc.1993.375
39	Wang Y. ; Zhou K. ; Huang G. ; Hensley C. ; Huang X. ; Ma X. ; Zhao T. ; Sumer B. D. ; DeBerardinis R. J. ; Gao J Nat. Mater 2014, 13, 204. doi: 10.1038/NMAT3819
40	Li C. ; Xia J. A. ; Wei X. B. ; Yan H. H. ; Si Z. ; Ju S. H Adv. Funct. Mater 2010, 20, 2222. doi: 10.1002/adfm.201000038
41	Li L. ; Ji J. ; Fei R. ; Wang C. ; Lu Q. ; Zhang J. ; Jiang L. ; Zhu J Adv. Funct. Mater 2012, 22, 2971. doi: 10.1002/adfm.201200166
42	Zheng X. ; Than A. ; Ananthanaraya A. ; Kim D. ; Chen P ACS Nano 2013, 7, 6278. doi: 10.1021/nn4023137
43	Zhou K. J. ; Liu H. M. ; Zhang S.R. ; Huang X. N. ; Wang Y. G. ; Huang G ; Sumer B. D. ; Gao J. M J. Am. Chem. Soc 2012, 134, 7803. doi: 10.1021/ja300176w
44	Hu C. ; Mu Y. ; Li M. ; Qiu J Acta Phys. -Chim. Sin 2019, 35 (6), 572.
	胡超; 穆野; 李明宇; 邱介山. 物理化学学报, 2019, 35 (6), 572. doi: 10.3866/PKU.WHXB201806060
45	Zhu S. ; Zhang J ; Qiao C. ; Tang S. ; Li Y. ; Yuan W. ; Li B. ; Tian L. ; Liu F. ; Hu R. ; et al Chem. Commun 2011, 47, 6858. doi: 10.1039/C1CC11122A
46	Jain R. K Cancer Metast. Rev 1987, 6, 559. doi: 10.1007/BF00047468

[1]	杨健, 雷辰, 刘祥, 张建, 孙玉蝶, 张铖, 叶明富, 张奎. 碳量子点阳离子表面活性剂的多功能性[J]. 物理化学学报, 2022, 38(12): 2111030 - .
[2]	黄彦捷, 连超, 周瑾艳, 黄梓宸, 康晓红, 黄振宇, 李小菁, 陈玲, 关妍. 由蚕砂制备的碳量子点在不同激发、pH、金属离子、温度及极性环境下的荧光性质研究[J]. 物理化学学报, 2019, 35(11): 1267 -1275 .
[3]	李少海,翁波,卢康强,徐艺军. 聚胺界面修饰改善碳量子点可见光光敏化性能[J]. 物理化学学报, 2018, 34(6): 708 -718 .
[4]	贺平,袁方龙,王子飞,谭占鳌,范楼珍. 基于碳量子点的光电器件应用新进展[J]. 物理化学学报, 2018, 34(11): 1250 -1263 .
[5]	申迎华, 刘慧敏, 李国卿, 翟志国, 树学峰, 台秀梅. pH响应型P(HEMA/MAA)纳米微凝胶分散液的凝胶化行为和流变性能[J]. 物理化学学报, 2011, 27(08): 1919 -1925 .
[6]	方云, 赖中宇, 庞萍萍, 江明. 部分水解聚丙烯酰胺-羟乙基纤维素的水相pH响应性自组装[J]. 物理化学学报, 2011, 27(07): 1712 -1718 .
[7]	赵云龙, 段炼, 乔娟, 张德强, 王立铎, 邱勇. 基于双极性小分子的单层非掺杂红色荧光有机发光二极管[J]. 物理化学学报, 2010, 26(03): 531 -534 .
[8]	曹发斌;田彦文;陈永杰;肖林久;刘云义. 电荷补偿对红色LED用荧光粉体的荧光性能改进[J]. 物理化学学报, 2009, 25(02): 299 -303 .