CD90^+肝癌干细胞的鉴定及靶向治疗研究

Identification and target therapy of CD90^+ liver cancer stem cells

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 detect the CD90~＋cells＇ stem cell characteristics in human hepatocellular carcinoma and BEL-7404 cell lines and killed CD90~＋liver cancer stem cells（LCSCs） by an anti-CD90 monoclonal antibody targeted thermosensitive magnetoliposomes encapsulated HSP90 inhibition 17-AA G（CD90@17-AAG / TMs）. Methods：Immunohistochemical staining was performed to explore the relationship between the expression of CD90 and the clinicopathological parameters of human hepatocellular carcinoma. Magnetic activated cell sorting was performed to isolate the CD90~＋cells from the BEL-7404 cell lines and its stem cell characteristics were identified. CD90@17-AAG / TMs was prepared by rotary evaporator-hydration method to kill the CD90＋LCSCs in vivo. Results：There was no significant correlation among the age,sex,tumor size,and AFP with the expression of CD90 in hepatocellular carcinoma tissues（P〈0.05）. The expression of CD90 was relative to the pathology classification（P〈0.05）. The CD90＋LCSCs showed a significantly stem cell characteristics. CD90@17-AAG / TMs effectively killed CD90＋LCSCs and inhibited the tumor growth. Conclusion：CD90＋LCSCs has significant stem cell characteristics. CD90@17-AAG / TMs can effectively kill CD90＋LCSCs in vivo and inhibit the formation and growth of tumors,which may provide a novel therapy approach for hepatocellular carcinoma and other cancers.