MiR-34a对卵巢癌肿瘤干细胞生物学特性的影响

发布时间：2018-05-21 20:06

本文选题：肿瘤干细胞 + 卵巢癌　；参考：《首都医科大学》2014年硕士论文

【摘要】：目的：探讨MiR-34a在卵巢癌肿瘤干细胞中的表达及其对人卵巢癌干细胞的生物学特性的影响。方法：本研究选择人卵巢上皮性肿瘤细胞株A2780作为样本，以CD133作为Marker，，采用流式细胞仪分选出CD133+、CD133-细胞，采用克隆实验方法验证CD133+细胞具有卵巢癌干细胞的生物学特性，采用RT-PCR方法比较验证miR-34a在CD133+细胞的表达。将miR-34a重组质粒转染给CD133+细胞，观察卵巢癌肿瘤干细胞生物学性状的改变，包括采用MTT法检测增殖能力和化疗敏感性，皮下接种裸鼠成瘤检测成瘤能力。结果：培养14天后，CD133+细胞的克隆形成能力显著高于CD133-细胞(P0．01)，且CD133+细胞能形成更大的克隆；miR-34a在CD133+细胞中低表达；培养1、2、3、4、5、6d后，转染miR-34a CD133+细胞与未转染miR-34a CD133+细胞（对照组）相比，细胞增殖能力降低(P0．05)；紫杉醇低至高浓度作用下，转染miR-34a CD133+细胞的耐药性低于对照组，细胞药物敏感性差异具有显著统计学意义(P0．01)；转染miR-34a CD133+细胞较未转染miR-34a CD133+细胞致瘤能力显著降低，相同数量的未转染miR-34a CD133+细胞在同一观察时间点可以形成更大的肿瘤，且形成肿瘤的潜伏期较短。结论：1、CD133可以作为卵巢癌肿瘤干细胞表面的标志性分子之一；2、miR-34a在卵巢癌肿瘤干细胞中低表达；3、miR-34a在卵巢癌中发挥潜在的抑癌基因作用，增加miR-34a的表达，可明显抑制细胞的增殖、成瘤能力及耐药性，大大降低了肿瘤细胞的恶性度；4、miR-34a可成为一个治疗卵巢癌的潜在基因靶点，可能为卵巢癌的治疗提供一条新的生物途径。
[Abstract]:Aim: to investigate the expression of MiR-34a in ovarian cancer stem cells and its effect on the biological characteristics of human ovarian cancer stem cells. Methods: in this study, human ovarian epithelial tumor cell line A2780 was selected as sample, CD133 as marker, CD133 CD133- cells were separated by flow cytometry, and the biological characteristics of ovarian cancer stem cells were verified by clone assay. The expression of miR-34a in CD133 cells was compared by RT-PCR method. The miR-34a recombinant plasmid was transfected into CD133 cells to observe the changes of biological characteristics of ovarian cancer stem cells. The proliferation and chemosensitivity of ovarian cancer stem cells were detected by MTT method and tumorigenic ability was detected by subcutaneous inoculation of nude mice. Results: after 14 days of culture, the clone forming ability of CD133 cells was significantly higher than that of CD133- cells, and CD133 cells could form a larger clone of miR-34a in CD133 cells, and after 6 days of culture, the transfected miR-34a CD133 cells were compared with those of untransfected miR-34a CD133 cells (control group). The drug resistance of miR-34a CD133 cells transfected with paclitaxel at low to high concentration was lower than that in control group. The chemosensitivity of miR-34a CD133 cells was significantly lower than that of untransfected miR-34a CD133 cells, and the same number of untransfected miR-34a CD133 cells could form larger tumors at the same observation time point. And the incubation period of tumor formation is short. Conclusion 1: 1 + CD133 can play a potential role as a tumor suppressor gene in ovarian cancer cells, increase the expression of miR-34a, and inhibit the proliferation of ovarian cancer cells, which is one of the signature molecules on the surface of ovarian cancer stem cells. The ability of tumorigenesis and drug resistance have greatly reduced the malignancy of tumor cells. 4miR-34a may become a potential gene target for the treatment of ovarian cancer and may provide a new biological pathway for the treatment of ovarian cancer.
【学位授予单位】：首都医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

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