目的 探讨鳖甲煎丸对裸鼠皮下成瘤、肝癌干细胞(liver cancer stem cells,LCSCs)干性的抑制作用及其相关机制.方法 选取12只BALB/c裸鼠,随机分为4组,即肝癌干细胞皮下成瘤+生理盐水灌胃组(A组)、肝癌干细胞皮下成瘤+鳖甲煎丸灌胃组(B组)、miR-140过表达慢病毒肝癌干细胞皮下成瘤+生理盐水灌胃(C组)、miR-140干扰慢病毒肝癌干细胞皮下成瘤+鳖甲煎丸灌胃(D组),将分选出肝癌干细胞种植于裸鼠皮下形成瘤体,记录瘤体体积大小,并进行多组间对比.苏木精-伊红(HE)染色观察瘤体组织的病理学变化.PCR和Western Bolt分别检测肝癌组织中CD133、CD24、EpCAM、AFP的mRNA表达、蛋白表达以及miR-140的基因表达.结果 B组、C组瘤体体积均小于A组,D组瘤体体积大于B组;HE染色结果提示瘤体组织呈恶性后进行PCR与Western Bolt检测,结果提示B组、C组的CD133、CD24、EpCAM、AFP的mRNA以及蛋白表达相对含量均小于A组,D组以上检测指标小于B组;予鳖甲煎丸灌胃后,瘤体中miR-140的表达升高.结论 鳖甲煎丸能抑制肝癌干细胞的致瘤能力以及降低肝癌组织中CD133、CD24、EpCAM、AFP,其作用机制可能与促进LCSCs中miR-140的表达有关.
Abstract
Objective To investigate the inhibitory effect of Biejia Jianwan on subcutaneous tumorigenesis and liver cancer stem cells(LCSCs)dryness in nude mice and its related mechanisms.Methods Twelve BALB/c nude mice were selected and randomly divided into four groups:subcutaneous tumorigenesis of liver cancer stem cells + saline gavage group(Group A),subcutaneous tumorigenesis of liver cancer stem cells + Biejia Jianwan gavage group(Group B),subcutaneous tumorigenesis of miR-140 overexpressing lentivirus liver cancer stem cells + saline gavage group(Group C),and miR-140 interference lentivirus liver cancer stem cells subcutaneous tumorigenesis + Biejia Jianwan gavage group(Group D).The selected liver cancer stem cells were subcutaneously planted in nude mice to form a tumor body,and the volumes of the tumors were recorded and compared among the groups.Hematoxylin eosin(HE)staining was used to observe the pathological changes of tumor tissue,while PCR and Western Bolt were used to detect the mRNA expressions,protein expressions of CD133,CD24,EpCAM,and AFP,and the gene expression of miR-140 in liver cancer tissue,respectively.Results The tumor volumes of Group B and Group C were smaller than that of Group A,while the tumor volume of Group D was larger than that of Group B.The HE staining results showed that the tumor tissue was malignant,and then the PCR and Western Bolt detections were performed.The results showed that the relative mRNA and protein expression levels of CD133,CD24,EpCAM,and AFP in Group B and Group C were lower than those in Group A,while the above detection indicators in Group D were lower than those in Group B.After oral administration of Biejia Jianwan,the expressions of miR-140 in tumors were increased.Conclusion Biejia Jianwan can inhibit the tumorigenic ability of liver cancer stem cells and reduce CD133,CD24,EpCAM,and AFP in liver cancer tissue.Its mechanism of action may be related to promoting the expression of miR-140 in LCSCs.
关键词
鳖甲煎丸/miR-140/肝癌干细胞/皮下成瘤
Key words
Biejia Jianwan/MiR-140/liver cancer stem cells/subcutaneous tumorigenesis
维普
万方数据