【摘要】：目的:觀察JQ1對(duì)宮頸癌HeLa細(xì)胞增殖的影響,探討JQ1對(duì)宮頸癌HeLa細(xì)胞增殖的調(diào)控作用及其機(jī)制,為臨床治療宮頸癌提供新的治療途徑。方法:1.體外培養(yǎng)HeLa細(xì)胞,按每孔細(xì)胞0.5×104個(gè)接種至96孔板,細(xì)胞貼壁100min后,將細(xì)胞分成二甲亞砜(DMSO)對(duì)照組和JQ1處理組(終濃度分別為0.01、0.1、1和10μmol/L),細(xì)胞處理96h,采用MTT比色法檢測(cè)JQ1對(duì)HeLa細(xì)胞增殖的影響;2.體外培養(yǎng)HeLa細(xì)胞,按每孔細(xì)胞500個(gè)接種至6孔板,細(xì)胞貼壁100min后,將細(xì)胞分成DMSO對(duì)照組和JQ1處理組(終濃度分別為0.02、0.2、2和20μmol/L),細(xì)胞處理12 d,計(jì)算細(xì)胞克隆形成數(shù)量變化;3.體外培養(yǎng)HeLa細(xì)胞,將細(xì)胞分成DMSO對(duì)照組和JQ1處理組(終濃度分別為0.01、0.1、1和10μmol/L),收集細(xì)胞RNA和蛋白,采用實(shí)時(shí)熒光定量PCR和Westernblot檢測(cè)細(xì)胞中c-Myc表達(dá)變化。結(jié)果:1.與DMSO對(duì)照組相比,JQ1抑制了HeLa細(xì)胞的增殖,且成劑量依賴性,0.01、0.1、1和10μmol/L JQ1處理組細(xì)胞增殖分別下降至0.8±0.02、0.71±0.1、0.61±0.01和0.45±0.02,均P0.05;2.與DMSO對(duì)照組相比,0.01、0.1、1和10μmol/L JQ1處理組均抑制了HeLa細(xì)胞克隆形成(細(xì)胞克隆形成數(shù)量:276.7±12比156±11、111.7±10、80.3±6、4.3±2,均P0.05);3.與DMSO對(duì)照組相比,0.01、0.1、1和10μmol/L JQ1處理組抑制了HeLa細(xì)胞c-Myc m RNA(1.01±0.01倍比0.74±0.02倍、0.63±0.021倍、0.52±0.01倍、0.45±0.02倍,均P0.05)和蛋白表達(dá)(1.02±0.01倍比0.64±0.15倍、0.53±0.02倍、0.42±0.11倍、0.33±0.02倍,均P0.05)。結(jié)論:JQ1抑制了HeLa細(xì)胞增殖,其可能的機(jī)制是通過抑制c-Myc的表達(dá)。
[Abstract]:Aim: to observe the effect of JQ1 on the proliferation of cervical cancer HeLa cells, and to explore the regulatory effect and mechanism of JQ1 on the proliferation of cervical cancer HeLa cells, so as to provide a new therapeutic approach for clinical treatment of cervical cancer. Methods: 1. HeLa cells were cultured in vitro. The cells were inoculated to 96 well plates according to 0.5 脳 104 cells per well. The cells were divided into dimethyl sulfoxide (DMSO) control group and JQ1 treated group (0. 01 渭 mol/L and 0. 01 渭 mol/L, respectively). The cells were treated for 96 h. The effect of JQ1 on the proliferation of HeLa cells was detected by MTT colorimetry. 2. HeLa cells were cultured in vitro. The cells were inoculated to 6 well plates according to 500 cells per well. After the cells adhered to 100min, the cells were divided into two groups: DMSO control group and JQ1 treatment group (the final concentration was 0.02 渭 mol/L and 20 渭 mol/L, respectively), and the cells were treated for 12 days. The number of cell clone formation was calculated. 3. HeLa cells were cultured in vitro. The cells were divided into DMSO control group and JQ1 treated group (the final concentration was 0.01 渭 mol/L and 10 渭 mol/L, respectively). The cell RNA and protein were collected and the changes of c-Myc expression in the cells were detected by real-time fluorescence quantitative PCR and Westernblot. The result is 1: 1. Compared with DMSO control group, JQ1 inhibited the proliferation of HeLa cells in a dose-dependent manner. The proliferation of HeLa cells in the 0. 01 渭 mol/L JQ1 and 10 渭 mol/L JQ1 groups decreased to 0. 8 鹵0. 02 鹵0. 71 鹵0. 01 鹵0. 61 鹵0. 01 and 0. 45 鹵0. 02, respectively (P 0. 05). Compared with DMSO control group, both 0. 01 渭 mol/L JQ1 and 10 渭 mol/L JQ1 treatment group inhibited HeLa cell clone formation (276.7 鹵12 vs 156 鹵11111.7 鹵10 0. 3 鹵64.3 鹵2, P0.05). Compared with the control group of DMSO, the c-Myc m RNA (of HeLa cells was inhibited by 0.01 鹵0.01,0.63 鹵0.021, 0.52 鹵0.01,0.45 鹵0.02,0.74 鹵0.02,0.63 鹵0.021, 0.52 鹵0.01,0.45 鹵0.02 times respectively in the 0. 01 鹵0. 01 and 10 渭 mol/L JQ1 groups. Protein expression (1.02 鹵0.01 times vs 0.64 鹵0.15 times, 0.53 鹵0.02 fold, 0.42 鹵0.11 fold, 0.33 鹵0.02 fold, P0.05). Conclusion: JQ1 inhibits the proliferation of HeLa cells by inhibiting the expression of c-Myc.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.33

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