TiIV uptake and release by human serum transferrin and recognition of TiIV-transferrin by cancer cells:: Understanding the mechanism of action of the anticancer drug titanocene dichloride

被引:226
作者
Guo, ML
Sun, HZ
McArdle, HJ
Gambling, L
Sadler, PJ [1 ]
机构
[1] Univ Edinburgh, Dept Chem, Edinburgh EH9 3JJ, Midlothian, Scotland
[2] Univ Hong Kong, Dept Chem, Hong Kong, Hong Kong, Peoples R China
[3] Rowett Res Inst, Div Nutr Pregnancy & Dev, Aberdeen AB21 9SB, Scotland
关键词
D O I
10.1021/bi000798z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The organometallic anticancer agent titanocene dichloride, Cp2TiCl2, is now in phase II clinical trials as an anticancer drug, but its mechanism of action is poorly understood. We show here that the interactions of Cp2TiCl2 with human serum transferrin (hTF) and that of Ti-2-hTF with adenosine triphosphate (ATP) have characteristics that could allow; transferrin to act as a mediator for titanium delivery to tumor cells. Such reactions may therefore be important to the anticancer activity of this new class of drugs. Cp2TiCl2 reacts rapidly with human apo-transferrin under physiological conditions (100 mM NaCl, 25 mM bicarbonate, and 4 mM phosphate, pH 7.4) with carbonate as a synergistic anion. The Cp ligands are released from the drug. Two-dimensional [H-1, C-13] NMR studies of epsilon-[C-13]Met-hTF show that Ti-IV loads the C-lobe first followed by the N-lobe and binds in the specific Fe-III sites. The protein conformational changes induced by Ti-IV appear to be similar to those induced by Fem. Carbonate can act as a synergistic anion in Ti-2-hTF but does not appear to be essential. A specific Ti-IV-hTF adduct is formed even in the absence of bicarbonate. When the pH of Ti-2-hTF solutions is lowered, no Ti-IV is released at the endosomal pH of ca. 5.0-5.5, but one Ti-IV dissociates between pH 4.5-2.0. In contrast, in the presence of 1 mM ATP, all Ti-IV is readily released from both lobes when the pH is lowered from 7.0 to 4.5. Moreover, FeIII displaces Ti-IV rapidly from the C-lobe of Ti-2-hTF (<5 min) but only slowly (days) from the N-lobe. Thus, the species FeCTiN-hTF might also provide a route for Ti-IV entry into tumor cells via the transferrin receptor. Ti-2-hTF effectively blocked cell uptake of radiolabeled Fe-59-hTF into BeWo cells, a human placental choriocarcinoma cell line in culture. These results imply that titanium transferrin might be recognized by the transferrin receptor and be taken up into cancer cells.
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页码:10023 / 10033
页数:11
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