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目的合成奥曲肽紫杉醇偶联物(paclitaxel-octreotide conjugates),探讨偶联物对人小细胞肺癌(SCLC)细胞株H446的细胞毒性和靶向性。方法合成分离并纯化奥曲肽紫杉醇偶联物PTX-OCT、2PTX-OCT、PTX-Tyr-OCT和2PTX-Tyr-OCT,实时定量PCR法检测人SCLC细胞株H446及人成纤维细胞中生长抑素受体(SSTR)的表达,四甲基偶氮唑蓝微量酶反应比色(MTT)法检测不同浓度和时间梯度的偶联药物对H446及人成纤维细胞的抑制率,流式细胞术检测1000nmol/L紫杉醇、PTX-OCT、2PTX-OCT、PTX-Tyr-OCT和2PTX-Tyr-OCT作用后H446细胞周期和凋亡率的变化。结果 H446细胞中SSTR1～5 mRNA均有较高量的表达,而人成纤维细胞中未检测到SSTR mRNA的表达。奥曲肽紫杉醇偶联物对H446细胞的杀伤作用与紫杉醇相似,具有浓度(1～1000 nmol/L)和时间(24、48、72h)依赖性;而对SSTR阴性的成纤维细胞,偶联药物的毒性作用要明显低于紫杉醇(P<0.05)。流式细胞术分析PTX-OCT、2PTX-OCT、PTX-Tyr-OCT和2PTX-Tyr-OCT作用后,H446细胞周期均停滞在G2/M期。结论奥曲肽紫杉醇偶联药物具有较好的靶向性,在SCLC的治疗中具有重要的应用前景。
Objective To synthesize paclitaxel-octreotide conjugates and investigate the cytotoxicity and targeting of the conjugates to human small cell lung cancer (SCLC) cell line H446. METHODS: PTX-OCT, 2PTX-OCT, PTX-Tyr-OCT and 2PTX-Tyr-OCT were isolated and purified. The real-time quantitative PCR was used to detect somatostatin in human SCLC cell line H446 and human fibroblasts (SSTR), MTT assay was used to detect the inhibition rate of H446 and human fibroblasts with different concentration and time gradient of conjugated drugs. Flow cytometry was used to detect the inhibitory effect of 1000 nmol / L paclitaxel, PTX-OCT, 2PTX-OCT, PTX-Tyr-OCT and 2PTX-Tyr-OCT cell cycle and apoptosis rate of H446 cells. Results SSTR1 ~ 5 mRNA was highly expressed in H446 cells, but not in human fibroblasts. Octreotide paclitaxel conjugate has the same killing effect on H446 cells as paclitaxel, and has a concentration (1-1000 nmol / L) and time (24, 48, 72h) dependence. For SSTR-negative fibroblasts, Toxicity was significantly lower than paclitaxel (P <0.05). The cell cycle of H446 cells was arrested in G2 / M phase after PTX-OCT, 2PTX-OCT, PTX-Tyr-OCT and 2PTX-Tyr-OCT were analyzed by flow cytometry. Conclusion Octreotide paclitaxel-conjugated drugs have good targeting and have important application prospects in the treatment of SCLC.