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        免疫毒素LHRHPE40對HeLa細胞表面硬度的影響

        2014-07-10 21:19:11張晶
        分析化學 2014年6期
        關鍵詞:癌細胞硬度表面

        張晶

        摘 要 利用基于原子力顯微鏡(AFM)的力譜技術(shù), 在正常生長的單個活細胞表面上, 實時動態(tài)地研究了免疫毒素HeLa細胞表面硬度的影響。采用HertzSneddon模型計算所得力曲線相應的楊氏模量。實驗表明,

        1 引 言

        不斷發(fā)展的納米技術(shù)使得科研工作者可以在納米級尺度上操縱生物分子和細胞,同時在pN(皮牛)級別上獲得各種分子間的相互作用力[1~3]。在這些納米技術(shù)中,原子力顯微鏡(AFM)是一項具有多種功能的技術(shù)手段。AFM可以在準生理條件下對生物樣品進行直接測量,不需要復雜的樣品處理過程,這使其迅速地被應用于各種生物樣品的研究中[4~7]。

        在過去的十多年中,研究者應用傳統(tǒng)分子生物學實驗方法對腫瘤細胞的生化性質(zhì)進行了大量研究。但是,腫瘤細胞的機械硬度一直都被忽視,盡管腫瘤組織的入侵過程和腫瘤細胞的表面硬度有密切關系[8,9]。近年,細胞硬度作為腫瘤治療中的一項潛在的生物物理指標受到越來越多的關注[10,11]。在單細胞水平上分析細胞的表面硬度對理解腫瘤組織對化療藥物的反應和評價腫瘤的預后效果十分重要[12]。1992年,Tao等應用AFM研究了被切片的生物組織的硬度[13]。Hoh等應用AFM研究了活細胞的表面硬度[14]。基于這些開創(chuàng)性的研究工作,越來越多的研究者致力于探測在各種不同生理條件下活細胞硬度的變化。Kloxin等通過改變基底成分分析了基底機械性能對心臟瓣膜間質(zhì)細胞活化成肌成纖維細胞的影響,對于組織再生工程合理地設計移植材料有著重要的意義[15]。Cross等用AFM測量了多種癌癥患者胸膜轉(zhuǎn)移癌細胞的硬度,發(fā)現(xiàn)轉(zhuǎn)移癌細胞的細胞硬度比良性細胞軟70%,為腫瘤的診斷和治療提供了依據(jù)[16]。

        免疫毒素LHRHPE40是一種高特異性的針對大量表達LHRH受體(LHRHR)的癌細胞組織的抗癌藥物。LHRHPE40的作用機理是:導向部分配體LHRH通過其自身的特異性識別能力與目標組織細胞表面的LHRHR結(jié)合,毒性部分PE40通過跨膜轉(zhuǎn)運的方式進入細胞內(nèi),發(fā)揮其藥效,殺死腫瘤細胞[17]。

        本研究采用AFM技術(shù)實時動態(tài)研究了LHRHPE40對HeLa細胞的硬度的影響,并且初步探索了引起HeLa細胞硬度發(fā)生變化的原因。

        2 實驗部分

        2.1 儀器與試劑

        4 結(jié) 論

        本研究應用AFM力譜技術(shù)考察了免疫毒素LHRHPE40對HeLa細胞的表面硬度的影響。結(jié)果表明,LHRHPE40會造成HeLa細胞在凋亡的過程中表面硬度逐步增加。熒光實驗結(jié)果表明, HeLa細胞硬度的增加與胞內(nèi)微絲骨架的重組聚集有關。這些實驗結(jié)果為從細胞表面硬度角度掌握LHRHPE40的藥用效果和作用機理提供了重要信息。但是,在LHRHPE40的作用過程中,細胞內(nèi)的各種生化組分發(fā)生了何種變化,仍需要借助其它如拉曼光譜等實驗技術(shù)的細胞成分分析功能進行進一步研究。

        References

        1 Neuman K C, Nagy A. Nat. Methods, 2008, 5(6): 491-505

        2 Dickenson N E, Armendariz K P, Huckabay H A, Livanec P W, Dunn R C. Anal. Bioanal. Chem., 2010, 396(1): 31-43

        3 JI TianRong, LIANG ZhongWei, ZHU XinYu, SHAO YuanHua. Chinese J. Anal. Chem., 2010, 38(12): 1821-1827

        紀天容, 梁中偉, 朱新宇, 邵元華. 分析化學, 2010, 38(12): 1821-1827

        4 Muller D J, Dufrene Y F. Nat. Nanotechnol., 2008, 3(5): 261-269

        5 Jiang J G, Hao X, Cai M J, Shan Y P, Shang X, Tang Z Y, Wang H D . Nano Lett., 2009, 9(12): 4489-4493

        6 Zhu R, Rupprecht A, Pohl E E. J. Am. Chem. Soc., 2013, 135(9): 3640-3646

        7 Shi X L, Zhang X J, Xia T, Fang X H. Nanomedicine, 2012, 7(10): 1625-1637

        8 Kumar S, Weaver V. Cancer Metast. Rev., 2009, 28(1): 113-127

        9 Mierke C T. Cell Biochem. Biophys., 2011, 61(2): 217-236

        10 Wilson L, Cross S, Gimzewski J, Rao J Y. Idrugs, 2010, 13(12): 847-851

        11 Suresh S. Nat. Nanotechnol., 2007, 2(12): 748-749

        12 Xiao L F, Tang M J, Li Q F, Zhou A H. Anal. MethodsUk, 2013, 5(4): 874-879

        13 Tao N J, Lindsay S M, Lees S. Biophys. J., 1992, 63(4): 1165-1169

        14 Hoh J H, Schoenenberger C A. J. Cell Sci., 1994, 107(5): 1105-1114

        15 Kloxin A M, Benton J A, Anseth K S. Biomaterials, 2010, 31(1): 1-8

        16 Cross S E, Jin Y S, Rao J, Gimzewski J K. Nat. Nanotechnol., 2007, 2(12): 780-783

        17 Deng X, Klussmann S, Wu G M, Akkerman D, Zhu Y Q, Liu Y, Chen H, Zhu P, Yu B Z, Zhang G L. J. Drug Target., 2008, 16: 379-388

        18 YE ZhiYi, ZHANG Li. Chinese Bull. Life Sci., 2010, 22(8): 817-822

        葉志義, 張 麗. 生命科學, 2010, 22(8): 817-822

        19 Domke J, Radmacher M. Langmuir, 1998, 14(12): 3320-3325

        Effect of Cancer Target Drug LHRHPE40 on Elasticity of HeLa Cells

        ZHANG Jing1,2, ZHANG BaiLin*1, TANG JiLin*1

        1 (State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Changchun 130022, China)

        2(University of Chinese Academy of Sciences, Beijing 100049, China)

        Abstract The quantitative analysis of biomechanical profiles at the singlecell level can provide additional information. It is usually not available in traditional cell biology approaches, but may be crucial to assess and understand tumor prognosis and response to chemotherapy. In this study, the online changes of cell elastic properties after the addition of cancer target drug LHRHPE40 were monitored by atomic force microscopy (AFM) on living HeLa cell surface under physiological condition. The results from AFM based force spectroscopy showed that LHRHPE40 induced a distinct increase of the cell surface elasticity of HeLa cells. The fluorescence images implied that the target drug LHRHPE40 would affect the reorganization of cell actions, which led to the increase of the elasticity of HeLa cells.

        Keywords Cell elasticity; Singlecell level; Force spectroscopy; Atomic force microscopy

        (Received 29 November 2013; accepted 23 March 2014)

        This work was supported by the National Natural Science Foundation of China (Nos. 20975096, 21075121, 21275140, 21375122) and the Major State Basic Research Development Program (No. 2011CB935800).

        15 Kloxin A M, Benton J A, Anseth K S. Biomaterials, 2010, 31(1): 1-8

        16 Cross S E, Jin Y S, Rao J, Gimzewski J K. Nat. Nanotechnol., 2007, 2(12): 780-783

        17 Deng X, Klussmann S, Wu G M, Akkerman D, Zhu Y Q, Liu Y, Chen H, Zhu P, Yu B Z, Zhang G L. J. Drug Target., 2008, 16: 379-388

        18 YE ZhiYi, ZHANG Li. Chinese Bull. Life Sci., 2010, 22(8): 817-822

        葉志義, 張 麗. 生命科學, 2010, 22(8): 817-822

        19 Domke J, Radmacher M. Langmuir, 1998, 14(12): 3320-3325

        Effect of Cancer Target Drug LHRHPE40 on Elasticity of HeLa Cells

        ZHANG Jing1,2, ZHANG BaiLin*1, TANG JiLin*1

        1 (State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Changchun 130022, China)

        2(University of Chinese Academy of Sciences, Beijing 100049, China)

        Abstract The quantitative analysis of biomechanical profiles at the singlecell level can provide additional information. It is usually not available in traditional cell biology approaches, but may be crucial to assess and understand tumor prognosis and response to chemotherapy. In this study, the online changes of cell elastic properties after the addition of cancer target drug LHRHPE40 were monitored by atomic force microscopy (AFM) on living HeLa cell surface under physiological condition. The results from AFM based force spectroscopy showed that LHRHPE40 induced a distinct increase of the cell surface elasticity of HeLa cells. The fluorescence images implied that the target drug LHRHPE40 would affect the reorganization of cell actions, which led to the increase of the elasticity of HeLa cells.

        Keywords Cell elasticity; Singlecell level; Force spectroscopy; Atomic force microscopy

        (Received 29 November 2013; accepted 23 March 2014)

        This work was supported by the National Natural Science Foundation of China (Nos. 20975096, 21075121, 21275140, 21375122) and the Major State Basic Research Development Program (No. 2011CB935800).

        15 Kloxin A M, Benton J A, Anseth K S. Biomaterials, 2010, 31(1): 1-8

        16 Cross S E, Jin Y S, Rao J, Gimzewski J K. Nat. Nanotechnol., 2007, 2(12): 780-783

        17 Deng X, Klussmann S, Wu G M, Akkerman D, Zhu Y Q, Liu Y, Chen H, Zhu P, Yu B Z, Zhang G L. J. Drug Target., 2008, 16: 379-388

        18 YE ZhiYi, ZHANG Li. Chinese Bull. Life Sci., 2010, 22(8): 817-822

        葉志義, 張 麗. 生命科學, 2010, 22(8): 817-822

        19 Domke J, Radmacher M. Langmuir, 1998, 14(12): 3320-3325

        Effect of Cancer Target Drug LHRHPE40 on Elasticity of HeLa Cells

        ZHANG Jing1,2, ZHANG BaiLin*1, TANG JiLin*1

        1 (State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Changchun 130022, China)

        2(University of Chinese Academy of Sciences, Beijing 100049, China)

        Abstract The quantitative analysis of biomechanical profiles at the singlecell level can provide additional information. It is usually not available in traditional cell biology approaches, but may be crucial to assess and understand tumor prognosis and response to chemotherapy. In this study, the online changes of cell elastic properties after the addition of cancer target drug LHRHPE40 were monitored by atomic force microscopy (AFM) on living HeLa cell surface under physiological condition. The results from AFM based force spectroscopy showed that LHRHPE40 induced a distinct increase of the cell surface elasticity of HeLa cells. The fluorescence images implied that the target drug LHRHPE40 would affect the reorganization of cell actions, which led to the increase of the elasticity of HeLa cells.

        Keywords Cell elasticity; Singlecell level; Force spectroscopy; Atomic force microscopy

        (Received 29 November 2013; accepted 23 March 2014)

        This work was supported by the National Natural Science Foundation of China (Nos. 20975096, 21075121, 21275140, 21375122) and the Major State Basic Research Development Program (No. 2011CB935800).

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