Buckwheat trypsin inhibitor enters Hep G2 cells by clathrin-dependent endocytosis

被引：18

作者：

Cui, Xiaodong ^{[1
]}

Wang, Zhuanhua ^{[1
]}

Li, Yuying ^{[1
]}

Li, Chen ^{[1
]}

机构：

[1] Shanxi Univ, Inst Biotechnol, Key Lab Chem Biol & Mol Engn, Minist Educ, Taiyuan 030006, Peoples R China

来源：

FOOD CHEMISTRY | 2013年 / 141卷 / 03期

关键词：

Cell penetration mechanism; Endocytosis; Buckwheat; Trypsin inhibitor; Drug delivery; BOWMAN-BIRK INHIBITOR; PROTEASE INHIBITORS; PROTEINASE-INHIBITORS; MEDIATED ENDOCYTOSIS; BREAST; SEEDS;

D O I：

10.1016/j.foodchem.2013.04.001

中图分类号：

O69 [应用化学];

学科分类号：

081704 ;

摘要：

Recombinant buckwheat trypsin inhibitor (rBTI) was studied to evaluate if it could enter cancer cells and to determine the mechanism. Fluorescein isothiocyanate-labelled buckwheat trypsin inhibitor (FITC-BTI) entered Hep G2 cells in a concentration-dependent manner. FITC-BTI colocalised with labelled transferrin (Tf) in the punctate structure, implying that rBTI enters Hep G2 cells by clathrin-dependent endocytosis. Incubation of Hep G2 cells with different chemical inhibitors abolished diffuse, but not punctate fluorescence, thus indicating that membrane potential plays a critical role in this process. Impairment of clathrin-mediated endocytosis by RNAi with clathrin heavy chain greatly reduced or completely abolished both diffuse and punctate fluorescence, further supporting a theory of a single route of endocytosis. Consistent with our working hypothesis, Hep G2 cells which were arrested in the M phase did not show any vesicular or diffuse FITC-BTI. We conclude from these results that both endocytosis and membrane potential are required for rBTI entry into Hep G2 cells. (C) 2013 Elsevier Ltd. All rights reserved.

引用

页码：2625 / 2633

页数：9

共 28 条

[1] DEPLETION OF INTRACELLULAR POTASSIUM DISRUPTS COATED PITS AND REVERSIBLY INHIBITS CELL POLARIZATION DURING FIBROBLAST SPREADING [J].

ALTANKOV, G ;

GRINNELL, F .

JOURNAL OF CELL BIOLOGY, 1993, 120 (06) :1449-1459

[2]

[Anonymous], [No title captured]

[3]

BELOZERSKY MA, 1995, FEBS LETT, V371, P264, DOI 10.1016/0014-5793(95)00899-K

[4] NATURAL PROTEIN PROTEINASE-INHIBITORS AND THEIR INTERACTION WITH PROTEINASES [J].

BODE, W ;

HUBER, R .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1992, 204 (02) :433-451

[5] Bowman-Birk inhibitor abates proteasome function and suppresses the proliferation of MCF7 breast cancer cells through accumulation of MAP kinase phosphatase-1 [J].

Chen, YW ;

Huang, SC ;

Lin-Shiau, SY ;

Lin, JK .

CARCINOGENESIS, 2005, 26 (07) :1296-1306

[6] Pea (Pisum sativum L.) protease inhibitors from the Bowman-Birk class influence the growth of human colorectal adenocarcinoma HT29 cells in vitro [J].

Clemente, A ;

Gee, JM ;

Johnson, IT ;

MacKenzie, DA ;

Domoney, C .

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2005, 53 (23) :8979-8986

[7]

De Leo F, 2002, NUCLEIC ACIDS RES, V30, P347, DOI 10.1093/nar/30.1.347

[8]

Dunaevsky YE, 1996, BIOCHEM MOL BIOL INT, V40, P199

[9] Penetration of protein toxins into cells [J].

Falnes, PO ;

Sandvig, K .

CURRENT OPINION IN CELL BIOLOGY, 2000, 12 (04) :407-413

[10] Predicting cell-penetrating peptides [J].

Hansen, Mats ;

Kilk, Kalle ;

Langel, Ulo .

ADVANCED DRUG DELIVERY REVIEWS, 2008, 60 (4-5) :572-579

← 1 2 3 →