New targeting therapy with complex of liposome, consist of comparable lipid composition of oral cancer cells, and anti-EGF receptor antibody.

很早以前就知道，抗癌药物的疗效取决于靶癌细胞的组织类型。我们用无血清培养法检测了5株人癌细胞对顺铂、多柔比星和紫杉醇的敏感性。在所测试的细胞系中，唾液腺癌细胞系(SAC)对顺铂和紫杉醇的敏感性普遍高于鳞状细胞癌细胞系(SCC)，而SCC对顺铂相对耐药。另一方面，鳞癌对培莱霉素和阿霉素的敏感性高于SAC。已知顺铂、培普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，这是由于癌细胞膜通透性的差异造成的。我们用…研究了无血清培养条件下细胞系的膜脂组成。更多的因素决定了膜的通透性。我们发现，鳞状细胞癌中总膜脂的70%是磷脂，其余的是游离胆固醇。另一方面，SAC中80%的膜脂是甘油三酯和酯化胆固醇等中性脂类，20%是磷脂。与SCC相比，SAC中性脂水平较高，可能导致膜流动性下降，顺铂积累量较高。而培普霉素和阿霉素对SCC有较强的细胞毒作用，其膜脂以磷脂为主，膜流动性高于SAC。这些结果提示癌细胞膜的脂质成分是决定癌细胞对顺铂、培铂、阿霉素和紫杉醇敏感性的主要因素。因此，我们设计了与鳞癌细胞膜的脂组成相当的脂质体，构建了脂质体包裹的顺铂或培铂，并在无血清培养中用生长实验检测了癌细胞和囊泡癌细胞对脂质体的敏感性。结果表明，与单独药物或药物-脂质体混合物相比，脂质体包裹的药物对SCC的细胞毒作用增强。此外，已知表皮生长因子(EGF)受体在鳞状细胞癌和囊状细胞癌上过度表达。因此，我们制备了抗EGF受体(12-93单抗)的抗体，并检测了CDDP与12-93单抗偶联的脂质体对NA和HSY的细胞毒作用。结果表明，与CDDP单独作用相比，该络合物对NA和HSY表现出更高的细胞毒性。较少

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, peplomycin doxorubicin and paclitaxel of five human cancer cell lines by growth assay in serum-free culture. Of the cell line tested, salivary gland adenocarcinoma cell lines(SAC) were shown to be generally more sensitive to cisplatinand paclitaxal than squamous cell carcnoma cell lines (SCC) in vitro, and SCC were relatively resistant to cisplatine. On the other hand SCC were more sensitive to peplomycin and doxorubicin in comparison to SAC. It is known that cisplatin, peplomycin and doxorubicin were uptaken in the cells by passive transport system. We have speculated that heterogeneity of these anti-cancer drug effects is correlated with intracellular drug levels, resulted from the difference of membrane permeability of cancer cells. We studied the membrane lipid composition of the cell lines in serum-free medium w … More hich determine the membrane permeability. We have found that 70% of total membrane lipid in SCC is phospholipid and remainder is free cholesterol. On the other hand, 80% of total membrane lipid in SAC is neutral lipid such as triglyceride and esterified cholesterol and 20% is phosholipid. The higher neutral lipid level of SAC which should have resulted in decreased membrane fluidity, is consistent with the higher accumulation of cisplatin compared SCC. On the other hand, peplomycin and doxorubicin exhibited high cytotoxicity to SCC, which membrane lipid consisted of phospholipid main]y and the membrane fluidity was higher than that of SAC. These results suggest that the lipid composition of cancer cell membrane is major factor determining the sensitivity of cancer cells to cisplatin, peplomycin, doxorubicin and paclrtaxel Thus we have designed liposome which is comparable to the lipid composition of SCC cell membrane, constructed the liposome-entrapped cisplatin or peplomycin and examined sensitivity to the liposome-entrapped drug of both SCC and SAC by growth assay in serum-free culture. As the result, the liposome-entrapped drug exhibited enhanced cytotoxicity on SCC compare to either drug alone or drug-liposome mixture. Furthermore it is known that the epidermal growth factor (EGF) receptor overexpress on SCC and SAC. So we prepared antibody to anti-EGF receptor (12-93 MoAb) and examined the cytotoxic effect of the CDDP-entapped liposome conjugated with 12-93 MoAb to NA and HSY. As result, this complex shown enhanced high cytotoxicity to NA and HSY compare CDDP alone. Less