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

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种人癌细胞系对顺铂、培霉素、阿霉素和紫杉醇的敏感性。在测试的细胞系中，涎腺腺癌细胞系（SAC）在体外对顺铂和紫杉醇的敏感性普遍高于鳞状细胞癌细胞系（SCC）， SCC对顺铂相对耐药。另一方面，与SAC相比，SCC对多霉素和阿霉素更敏感。已知顺铂、培霉素和阿霉素是通过被动转运系统在细胞内被摄取的。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通透性的差异引起的。我们在无血清培养基中研究了细胞系的膜脂组成。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%是中性脂，如甘油三酯和酯化胆固醇，20%是磷脂。SAC较高的中性脂质水平本应导致膜流动性下降，与SCC相比，顺铂的积累较高是一致的。另一方面，培霉素和阿霉素对SCC表现出较高的细胞毒性，SCC的膜脂主要由磷脂组成，膜流动性高于SAC。这些结果表明，癌细胞细胞膜的脂质组成是决定癌细胞对顺铂、培霉素、阿霉素和紫杉醇敏感性的主要因素，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或培霉素，并在无血清培养中通过生长试验检测了SCC和SAC对脂质体包裹的药物的敏感性。结果，脂质体包裹的药物与单独药物或药物-脂质体混合物相比，对SCC表现出增强的细胞毒性。此外，已知表皮生长因子（EGF）受体在SCC和SAC上过表达。因此，我们制备了抗egf受体（12-93 MoAb）抗体，并检测了cddp包埋的脂质体与12-93 MoAb偶联对NA和HSY的细胞毒性作用。结果表明，与单独CDDP相比，该复合物对NA和HSY表现出更高的细胞毒性。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