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目的 :对人肝癌组织和细胞株BEL-7404中的CD90~+细胞进行干细胞特性鉴定;制备载17-AAG的抗CD90磁性热敏脂质体(CD90@17-AAG/TMs)靶向杀伤CD90~+肝癌干细胞(liver cancer stem cells,LCSCs)。方法:利用免疫组化染色检测人肝癌组织中CD90的表达,并研究其与肝癌临床病理特征的关系;利用免疫磁珠分选法从人肝癌细胞株BEL-7404中分离出CD90~+LCSCs并验证其干细胞特性;“旋转蒸发-水化法”制备CD90@17-AAG/TMs,通过体内杀伤实验验证其杀伤CD90~+LCSCs的能力。结果:CD90的表达与患者性别、年龄、肿瘤大小、甲胎蛋白(AFP)无关(P>0.05),与病理分级有关(P<0.05);肝癌细胞中CD90~+细胞具有明显的干细胞特性;CD90@17-AAG/TMs在体内可以有效杀伤CD90~+LCSCs,抑制肝癌的生长。结论:CD90~+细胞具有明显的干细胞特性,采用CD90@17-AAG/TMs可有效清除CD90~+LCSCs,抑制肝癌的形成及生长,为肝癌及其他恶性肿瘤的治疗提供了新的手段。
OBJECTIVE: To identify stem cell characteristics of CD90 ~ + cells in human hepatocellular carcinoma cell line BEL-7404 and CD90 @ 17-AAG / TMs loaded with 17-AAG to kill CD90 ~ + Liver cancer stem cells (LCSCs). Methods: The expression of CD90 in human hepatocellular carcinoma (HCC) was detected by immunohistochemical staining and the relationship between the expression of CD90 and clinicopathological features was analyzed. CD90 ~ + LCSCs were isolated from human hepatocellular carcinoma cell line BEL-7404 by immunomagnetic bead sorting The ability of CD90 @ 17-AAG / TMs to kill CD90 ~ + LCSCs was tested by in vivo killing assay. Results: The expression of CD90 was not related to the gender, age, tumor size and AFP (P> 0.05), but correlated with the pathological grade (P <0.05). CD90 ~ + cells had obvious stem cell characteristics in hepatoma cells. CD90 @ 17-AAG / TMs can effectively kill CD90 ~ + LCSCs in vivo and inhibit the growth of liver cancer. CONCLUSION: CD90 ~ + cells have obvious stem cell characteristics. CD90 @ 17-AAG / TMs can effectively remove CD90 ~ + LCSCs and inhibit the formation and growth of hepatocellular carcinoma. It provides a new method for the treatment of liver cancer and other malignant tumors.