Department of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetspark 2, DK-2100 Copenhagen East, Denmark.
Metallomics. 2010 Dec;2(12):811-8. doi: 10.1039/c0mt00046a. Epub 2010 Nov 9.
The accumulation and cytotoxicity of a 10 μmol L⁻¹ equimolar human serum albumin-cisplatin adduct (HSA-Pt) was investigated in suspension Ehrlich Ascites Tumor Cells (EATC) and adherent Ehrlich Lettré Ascites Cells (Lettré). HSA-Pt did not induce apoptosis nor was it taken up by the cells to any significant amount within 24 h incubation. The accumulation and cytotoxicity of HSA-Pt was compared to 10 μmol L⁻¹ cisplatin for which a larger accumulation and cytotoxicity were observed in EATC compared to Lettré. The experiment was performed with cell medium exchange every fourth hour as HSA-Pt and cisplatin were not stable in RPMI-1640 with 10% serum. The stability was determined using size exclusion chromatography-inductively coupled plasma-mass spectrometry (SEC-ICP-MS) and after 4 h new platinum peaks were observed. These findings indicate that before conducting cell experiments, the stability of the compound in the cell medium should be investigated especially when long exposure times are applied. Furthermore, HSA-Pt was found to be stable in Hanks Balanced Saline Solution (HBSS) and in Phosphate Buffered Saline (PBS) at pH 5.3, 6.1 and 7.4. Thus, the shift in pH when HSA-cisplatin passes from blood (pH 7.4) to tumor tissue (pH 5-6) is not capable of releasing cisplatin from HSA.
研究了 10μmol L⁻¹等摩尔人血清白蛋白-顺铂加合物（HSA-Pt）在悬浮 Ehrlich 腹水肿瘤细胞（EATC）和贴壁 Ehrlich Lettré 腹水细胞（Lettré）中的积累和细胞毒性。在 24 小时孵育期内，HSA-Pt 既没有诱导细胞凋亡，也没有被细胞大量摄取。与 10μmol L⁻¹顺铂相比，HSA-Pt 的积累和细胞毒性进行了比较，发现顺铂在 EATC 中的积累和细胞毒性大于 Lettré。由于 HSA-Pt 和顺铂在含 10%血清的 RPMI-1640 中不稳定，因此每隔 4 小时进行一次细胞培养基交换，进行实验。使用尺寸排阻色谱-电感耦合等离子体质谱法（SEC-ICP-MS）测定稳定性，4 小时后观察到新的铂峰。这些发现表明，在进行细胞实验之前，应研究化合物在细胞培养基中的稳定性，尤其是在应用长时间暴露的情况下。此外，HSA-Pt 在 Hank's 平衡盐溶液（HBSS）和 pH 值为 5.3、6.1 和 7.4 的磷酸盐缓冲盐水（PBS）中稳定。因此，当 HSA-顺铂从血液（pH 值 7.4）转移到肿瘤组织（pH 值 5-6）时，pH 值的变化不能将顺铂从 HSA 中释放出来。
