Development of new drug deliverty system with liposome consist of comparable to the lipid composition of oral cancer cancer cell membrane

人们早就知道抗癌药物的作用依赖于靶癌细胞的组织学类型。我们用无血清培养法检测了5种人类癌细胞系对顺铂、阿霉素和培霉素的敏感性。在所测试的细胞系中，体外实验显示，粘液腺腺癌细胞系（SAC）通常比鳞状细胞癌细胞系（SCC）对顺铂更敏感，SCC对顺铂相对耐药。另一方面，SCC对阿霉素和培霉素的敏感性高于SAC。已知顺铂、培霉素和阿霉素是通过被动转运系统在细胞内摄取的。我们推测这些抗癌药物作用的异质性与细胞内药物水平有关，这是由癌细胞细胞膜透性的差异引起的。我们在无血清培养基中研究了细胞系的膜脂组成，这决定了细胞膜的通透性。我们发现SCC总膜脂的70%是磷脂，其余是游离胆固醇。另一方面，SAC总膜脂的80%为中性脂，如甘油三酯和酯化胆固醇，20%为磷脂。SAC较高的中性脂质水平本应导致膜流动性降低，与SCC相比，顺铂的积累较高是一致的。另一方面，阿霉素和培霉素对鳞状细胞癌表现出较高的细胞毒性，鳞状细胞癌的膜脂以磷脂为主，膜流动性高于SAC。这些结果提示，癌细胞细胞膜脂质组成是决定癌细胞对顺铂、培霉素和阿霉素敏感性的主要因素。因此，我们设计了与SCC细胞膜脂质组成相当的脂质体，构建了脂质体包裹的顺铂，并在无血清培养中通过生长试验检测了SCC和SAC对脂质体包裹的顺铂的敏感性。结果表明，与单独使用顺铂或顺铂-脂质体混合物相比，脂质体包裹的顺铂对SCC的细胞毒性增强。我们发现表皮生长因子受体（EGFR）在SCC和SAC上过表达。然后制备抗EGFR单克隆抗体（12-93），并用亲和素-生物素法联合脂质体包裹顺铂与抗EGFR单克隆抗体。我们在体外检测了SCC和SAC对联合抗egfr单克隆抗体的脂质体的敏感性。与脂质体包裹的顺铂相比，这种脂质体有增强SCC和SAC细胞毒性的倾向。少

It has long been known that the effect of anti-cancer drug is dependent on the histological type of the target cancer cells. We have examined the sensitivity to cisplatin, doxorubicin and peplomycin of five human cancer cell lines by growth assay in serum-free culture. Of the cell line tested, slivary gland adenocarcinoma cell lines (SAC) were shown to be generally more sensitive to cisplatin than squmous cell carcinoma cell lines (SCC) in vitro, and SCC were relatively resistant to cisplatin. On the other hand, SCC were more sensitive to doxorubicin and peplomycin in comparison to SAC.It is known that cisplatin, peplomycin and doxorubicin were uptake in the cells by passive transport system. We have speculated that heterogeneity of these anticancer drug effects is correlated with intracellular drug level, resulted from the difference of membrane permiability of cancer cells. We studied the membrane lipid composition of the cell lines in serum-free medium, which determine the membrane … More permiability. We have found that 70% of total membrane lipid in SCC is phosholipid and remainder is free cholesterol. On the other hand, 80% of total membrane lipid in SAC in neutral lipid such as triglyceride and esterified cholesterol, and 20% is phospholipid. The higher neutral lipid level of SAC which should have resulted in decreased membrane fluidity, is consistent with the higher accumulation of cisplatin compared to SCC.On the other hand, doxorubicin and peplomycin exhibited high cytotoxicity to SCC,which membrane lipid consisted of phospholipid mainly and the membrane fluidity was higher than that of SAC.These result suggest that the lipid composition of cancer cell membranes is major factor determining the sensitivity of cancer cells to cisplatin, peplomycin and doxorubicin. Thus, we have designed liposome which is comparable to the lipid composition of SCC cell membrane, constructed the liposome-entrapped cisplatin and examined sensitivity to the liposome-entrapped cisplatin of both SCC and SAC by growth assay in serum-free culture. As the reult, the liposome-entrapped cisplatin exhibited enhanced cytotoxicity on SCC compare to either cisplatin alone or cisplatin-liposome mixture. And we have shown that epidermal growth factor receptor (EGFR) was over-expression on SCC and SAC.Then we made monoclonal antibody (12-93) to EGFR and combined liposome-entrapped cisplatin with anti-EGFR monoclonal antibody with avidin-biotin method. We have examined sensitivity to the liposome united with anti-EGFR monoclonal antibody of both SCC and SAC in vitro. This liposome had a tendency to enhance cytotoxicity on SCC and SAC compare to liposome-entrapped cisplatin. Less