【摘要】 目的:通过噬菌体展示体内筛选技术已获得可特异性结合多种人肝细胞肝癌细胞系和肝癌肿瘤血管7 肽小分子LCI-X7。探讨其在体内、体外对肝癌的抑制作用。方法:采用MTT、流式细胞仪、BrdU染色、Boyden小室等方法分别研究LCI- X7对人肝细胞肝癌细胞系HCCLM6细胞的增殖、凋亡、细胞周期、运动及侵袭的影响,并初步探索其治疗剂量范围。采用LM6人肝癌裸鼠模型,研究腹腔注射LCI- X7对HCCLM6肿瘤生长及转移的影响,腹腔注射5 FU(50 mg·kg 1 ·周1)为阳性对照,相同体积生理盐水为阴性对照。检测肿瘤组织CD31抗体染色的微血管密度以观察LCI- X7治疗对肿瘤血管生成的影响。结果:与具有相同氨基酸不同序列的对照肽相比,LCI X7 在0.1×10-9M至1×109M浓度范围内未能改变HCCLM6 在体外的增殖和细胞周期;而1×10-6M LCI X7可减少约70%HCCLM6 细胞的穿膜运动(P<0.05)及侵袭(P<0.05);在裸鼠实验,LCI X7(500μg·kg 1)与对照相比,显著降低肝内及肺转移率,分别为1/10 vs. 7/10及4/10 vs. 10/10(P<0.05);肿瘤微血管染色的结果显示,LCI- X7治疗可降低微血管密度至51.3%(P<0.05)。LCI- X7 治疗对裸鼠无明显毒性。结论:LCI- X7可抑制HCCLM6的运动和侵袭,并可减少HCCLM6在裸鼠体内的肝内播散和肺转移。更多还原

【Abstract】 Objective: LCI-X7 is a 7-amino acid peptide, which was selected by in vivo phage display and specifically bound to several HCC cell lines and tumor vessels in hepatocellular carcinoma (HCC) in nude mice model. In this study, we tried to evaluate its in vitro and in vivo effects on HCC. Methods: The effects of LCI-X7 on proliferation, apoptosis, cell cycles, motility and invasiveness of HCCLM6 cell line were studied in vitro by MTT, flow cytometry, BrdU staining, Boyden chamber methods. Its effects on tumor growth and metastasis of HCCLM6 were also investigated in nude mice model. 5-FU(50mg·kg -1·w -1)was served as positive control. Microvessel density was calculated by CD31 antibody staining in tumor tissues. Results: LCI-X7 (1nM) inhibited motility and invasiveness of HCCLM6 by 3 folds in Boyden chamber assay (P<0.05); However, LCI-X7 did not affect other aspects of HCCLM6 at the tested dosage. In vivo study, LCI-X7 (500μg·kg -1) reduced the incidences of intrahepatic metastasis and lung metastasis compared with control (1/10 vs. 7/10 and 4/10 vs. 10/10, P< 0.05). LCI-X7 decreased microvessel density (6.7/field vs. 13.06/field at 200 magnified field under microscope, P< 0.05). Conclusion:LCI-X7 reduced motility and invasiveness of LM6 in vitro and inhibited intrahepatic and lung metastasis of LM6 in vivo.更多还原