靶向Wnt信号的溶瘤腺病毒抑制肝癌干细胞研究

发布时间：2018-01-29 15:07

本文关键词： 肝癌类干细胞溶瘤腺病毒 Wnt信号通路 TSLC　出处：《生物化学与生物物理进展》2017年04期 　论文类型：期刊论文

【摘要】：肝细胞癌(hepatocellular carcinoma,HCC)是全球第五大癌症并成为癌症死亡的主要原因,传统治疗早期肝癌取得了一定的进展,但是癌症的复发、转移和耐药仍未得到根本解决,这些现象可通过癌症干细胞理论(cancer stem cell,CSC)进行解释.本研究通过悬浮富集培养的方法,获得了MHCC-97H细胞的三维立体球细胞(sphere cell),并检测其干细胞特性,通过删除5型腺病毒的E1A CR2区域24 bp碱基,并用Wnt活性转录元件TCF/TEF调控E1A基因表达,同时插入抗癌基因TSLC1,得到了双靶向溶瘤腺病毒Ad.wnt-E1A(△24 bp)-TSLC1,通过MTT、结晶紫染色实验、Hoechst、细胞划痕、蛋白质印迹技术、Transwell及免疫荧光技术检测重组病毒对肝癌类干细胞的EMT(epithelial-mesenchymal transition)转化、杀伤、凋亡以及迁移的作用.结果表明,悬浮富集培养的MHCC-97H sphere细胞具有自我更新、分化能力、静息性以及耐药性,高表达肝癌干细胞表面标志物(如CD133等),重组病毒处理后表现出明显的杀伤效果及抑制细胞迁移与侵袭的特性,靶向抑制MHCC-97H sphere细胞能力更强(P0.001),且重组病毒能有效诱导肝癌类干细胞通过caspase途径发生凋亡.因此,重组病毒Ad.wnt-E1A(△24 bp)-TSLC1有可能成为靶向肝癌干细胞的治疗药物,具有一定的临床应用前景.
[Abstract]:Hepatocellular carcinoma (hepatocellular, carcinoma, HCC) is the world's fifth largest cancer and become the leading cause of cancer death, traditional treatment has made some progress, but the recurrence of cancer, metastasis and drug resistance have not been fundamentally solved, these phenomena can be through the cancer stem cell theory (cancer stem cell, CSC) this was explained. Through the research on the method of suspended enrichment culture, the three-dimensional sphere cells of MHCC-97H cells (sphere cell), and to detect the characteristics of stem cells, by removing the adenovirus type 5 E1A CR2 24 BP bases, and regulation of E1A gene expression by Wnt activated transcription element TCF/TEF, and inserted into the TSLC1 gene, obtained the double oncolytic adenovirus Ad.wnt-E1A (delta 24 BP) by MTT, -TSLC1, Hoechst, crystal violet staining experiments, cell scratch, Western blot, Transwell and immunofluorescence detection of recombinant virus Liver cancer stem cells EMT (epithelial-mesenchymal transition) transformation, killing, apoptosis and migration. The results show that the suspension of enrichment of MHCC-97H sphere cells with self-renewal, differentiation, and resting resistance, high expression of cancer stem cell surface markers (such as CD133), recombinant virus treatment showed characteristic killer the obvious effect and inhibition of cell migration and invasion, targeted inhibition of MHCC-97H sphere cells (P0.001), and the ability of recombinant virus can induce liver cancer stem cells through the caspase pathway to apoptosis. Therefore, recombinant virus Ad.wnt-E1A (delta 24 BP) -TSLC1 may be targeting liver cancer stem cell therapeutics, has a certain the prospect of clinical application.

【作者单位】：浙江理工大学生命科学学院;
【基金】：国家自然科学基金(81272687) 浙江省自然科学基金(LZ13H160004,LY16H160056) 浙江省大学生科技创新活动计划(2016R406007)资助项目~~
【分类号】：R735.7
【正文快照】： 根据《2015中国癌症统计》显示,中国2015年有429.2万例癌症新发病例以及281.4万例癌症死亡病例.随着发病率和死亡率的增加,癌症已经成为中国人群死亡的首要原因和主要的公共健康问题[1].肝癌是目前全球第五大常见的癌症,并成为癌症死亡的主要原因之一.目前通过手术、化疗以及

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