本文選題：腫瘤干細(xì)胞 + 卵巢癌　； 參考：《首都醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：探討MiR-34a在卵巢癌腫瘤干細(xì)胞中的表達(dá)及其對人卵巢癌干細(xì)胞的生物學(xué)特性的影響。方法：本研究選擇人卵巢上皮性腫瘤細(xì)胞株A2780作為樣本，以CD133作為Marker，，采用流式細(xì)胞儀分選出CD133+、CD133-細(xì)胞，采用克隆實驗方法驗證CD133+細(xì)胞具有卵巢癌干細(xì)胞的生物學(xué)特性，采用RT-PCR方法比較驗證miR-34a在CD133+細(xì)胞的表達(dá)。將miR-34a重組質(zhì)粒轉(zhuǎn)染給CD133+細(xì)胞，觀察卵巢癌腫瘤干細(xì)胞生物學(xué)性狀的改變，包括采用MTT法檢測增殖能力和化療敏感性，皮下接種裸鼠成瘤檢測成瘤能力。結(jié)果：培養(yǎng)14天后，CD133+細(xì)胞的克隆形成能力顯著高于CD133-細(xì)胞(P0．01)，且CD133+細(xì)胞能形成更大的克隆；miR-34a在CD133+細(xì)胞中低表達(dá)；培養(yǎng)1、2、3、4、5、6d后，轉(zhuǎn)染miR-34a CD133+細(xì)胞與未轉(zhuǎn)染miR-34a CD133+細(xì)胞（對照組）相比，細(xì)胞增殖能力降低(P0．05)；紫杉醇低至高濃度作用下，轉(zhuǎn)染miR-34a CD133+細(xì)胞的耐藥性低于對照組，細(xì)胞藥物敏感性差異具有顯著統(tǒng)計學(xué)意義(P0．01)；轉(zhuǎn)染miR-34a CD133+細(xì)胞較未轉(zhuǎn)染miR-34a CD133+細(xì)胞致瘤能力顯著降低，相同數(shù)量的未轉(zhuǎn)染miR-34a CD133+細(xì)胞在同一觀察時間點(diǎn)可以形成更大的腫瘤，且形成腫瘤的潛伏期較短。結(jié)論：1、CD133可以作為卵巢癌腫瘤干細(xì)胞表面的標(biāo)志性分子之一；2、miR-34a在卵巢癌腫瘤干細(xì)胞中低表達(dá)；3、miR-34a在卵巢癌中發(fā)揮潛在的抑癌基因作用，增加miR-34a的表達(dá)，可明顯抑制細(xì)胞的增殖、成瘤能力及耐藥性，大大降低了腫瘤細(xì)胞的惡性度；4、miR-34a可成為一個治療卵巢癌的潛在基因靶點(diǎn)，可能為卵巢癌的治療提供一條新的生物途徑。
[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.
【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.31

【共引文獻(xiàn)】

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1 張偉;四君子湯藥物血清對人胃癌細(xì)胞株BGC-823、SGC-7901側(cè)群細(xì)胞生物學(xué)特性的影響[D];蚌埠醫(yī)學(xué)院;2013年

本文編號：1920591