人绒癌干细胞样细胞的体内鉴定及其化疗耐药特性的初步研究

发布时间：2018-04-10 09:05

本文选题：肿瘤干细胞　切入点：绒毛膜癌　出处：《中南大学》2014年硕士论文

【摘要】：[目的]用无血清悬浮培养法从人绒毛膜癌JEG-3细胞系中分离培养绒毛膜癌干细胞样细胞,对其进行体内鉴定,并对其进行化疗药物敏感性的初步研究。 [方法](1)无血清培养基悬浮培养富集法分离培养JEG-3干细胞样细胞；(2)裸鼠体内成瘤实验观察JEG-3干细胞样细胞及JEG-3亲本细胞两组细胞的体内成瘤能力； (3) Transwell体外侵袭实验观察两组细胞的侵袭能力； (4) Real-time PCR检测两组细胞中OCT-4、NANOG的表达水平; (5)MTT法观察两组细胞在不同浓度下对依托泊苷、氟尿嘧啶和甲氨蝶呤敏感性的变化。 [结果] (1)JEG-3可在SFM中存活、增殖,形成悬浮生长具有自我更新能力的肿瘤干细胞样细胞,并可连续传代； (2)JEG-3干细胞样细胞较JEG-3亲本细胞具有更高致瘤性； (3)JEG-3干细胞样细胞透过膜的细胞较JEG-3亲本细胞多,JEG-3干细胞样细胞较JEG-3亲本细胞具有更强的侵袭能力(P值=0.006,P值0.05); (4)人绒癌JEG-3干细胞样细胞OCT-4、NANOG mRNA的表达均高于JEG-3亲本细胞,差异具有显著统计学意义(P0.05)； (5)在不同浓度的化疗药物作用下,人绒癌JEG-3干细胞样细胞比JEG-3细胞存活率更高,更具有耐药性,差异具有显著统计学意义(P0.05)。 [结论] 1.人绒毛膜癌JEG-3干细胞样细胞较JEG-3细胞具有更高的致瘤能力及侵袭能力； 2.人绒毛膜癌JEG-3干细胞样细胞对化疗药物具有显著耐药性；且其高表达OCT-4、NANOG,为绒癌的靶向治疗提供了靶点。
[Abstract]:[objective] to isolate and culture choriocarcinoma stem cell like cells from human choriocarcinoma JEG-3 cell lines by serum-free suspension culture, and to identify them in vivo and to study their chemosensitivity.[methods] the tumorigenic ability of JEG-3 stem cell like cells and JEG-3 parent cells was observed in nude mice by suspension culture enrichment method in serum-free medium.The invasion ability of the two groups was observed by Transwell in vitro.Real-time PCR was used to detect the expression of OCT-4 nano G in the two groups.The sensitivity of the two groups to etoposide, fluorouracil and methotrexate was observed by MTT assay.[results]JEG-3 could survive and proliferate in SFM and form tumor stem cell like cells with self-renewal ability, which could be subcultured continuously.JEG-3 stem cell like cells had higher tumorigenicity than JEG-3 parent cells.Compared with JEG-3 parent cells, JEG-3 stem cell like cells had stronger invasiveness than JEG-3 parent cells.The expression of NANOG mRNA in human choriocarcinoma JEG-3 stem cell like cells was significantly higher than that in JEG-3 parent cells, and the difference was statistically significant (P 0.05).The survival rate and drug resistance of human choriocarcinoma JEG-3 stem cell like cells were higher than those of JEG-3 cells under different concentrations of chemotherapeutic drugs, and the difference was statistically significant (P 0.05).[conclusion]1.JEG-3 stem cell like cells of human choriocarcinoma have higher tumorigenicity and invasiveness than JEG-3 cells.2.JEG-3 stem cell like cells of human choriocarcinoma have significant resistance to chemotherapeutic drugs, and their overexpression of OCT-4 NANOG provides a target for targeted treatment of choriocarcinoma.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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