發(fā)布時(shí)間：2018-03-06 04:35

  本文選題：宮頸癌細(xì)胞　切入點(diǎn)：ISG15　出處：《蘇州大學(xué)》2014年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：第一章ISG15通過(guò)抑制NF-B途徑誘導(dǎo)HeLa細(xì)胞的凋亡 目的：研究ISG15對(duì)宮頸癌細(xì)胞HeLa的促凋亡能力并探討其分子機(jī)制。 方法：(1)使用PCR技術(shù)克隆ISG15并構(gòu)建其重組質(zhì)粒及慢病毒載體；（2）過(guò)表達(dá)ISG15后，采用細(xì)胞計(jì)數(shù)法檢測(cè)ISG15對(duì)HeLa細(xì)胞增殖的影響；（3）采用Western blot技術(shù)檢測(cè)ISG15對(duì)HeLa細(xì)胞促凋亡的影響；（4）采用細(xì)胞劃痕實(shí)驗(yàn)檢測(cè)ISG15對(duì)HeLa細(xì)胞遷移的影響。 結(jié)果：（1）過(guò)表達(dá)ISG15能夠抑制HeLa細(xì)胞的增殖；(2)ISG15能夠通過(guò)抑制NF-B途徑誘導(dǎo)HeLa細(xì)胞的凋亡（；3）ISG15能夠抑制HeLa細(xì)胞的遷移。 結(jié)論：過(guò)表達(dá)ISG15能夠通過(guò)抑制NF-B途徑誘導(dǎo)HeLa細(xì)胞的凋亡。 第二章ISG15在氯碘羥喹誘導(dǎo)的血液病細(xì)胞凋亡中的作用研究 目的：對(duì)ISG15在氯碘羥喹抗血液病作用中的機(jī)制研究。 方法：（1）采用RT-PCR和Western blot技術(shù)檢測(cè)氯碘羥喹對(duì)血液病細(xì)胞中ISG15的mRNA和蛋白表達(dá)水平的影響；（2）采用RT-PCR技術(shù)檢測(cè)氯碘羥喹對(duì)干擾素信號(hào)通路的影響；（3）采用慢病毒介導(dǎo)血液病細(xì)胞過(guò)表達(dá)ISG15之后，，通過(guò)MTT法和Western blot技術(shù)檢測(cè)過(guò)表達(dá)的ISG15對(duì)血液病細(xì)胞的增殖和凋亡作用。 結(jié)果：（1）氯碘羥喹能夠顯著誘導(dǎo)血液病細(xì)胞中ISG15的表達(dá)，同時(shí)還能誘導(dǎo)其去結(jié)合酶USP18的表達(dá)；（2）氯碘羥喹能夠激活JAK-STAT信號(hào)通路從而誘導(dǎo)ISG15；（3）過(guò)表達(dá)ISG15能夠抑制血液病細(xì)胞的增殖并促進(jìn)其凋亡。 結(jié)論：氯碘羥喹能夠誘導(dǎo)ISG15的上調(diào)并且過(guò)表達(dá)ISG15能夠促進(jìn)血液病細(xì)胞的凋亡。
[Abstract]:Chapter 1 ISG15 induces apoptosis of HeLa cells by inhibiting NF-B pathway. Aim: to investigate the apoptotic effect of ISG15 on cervical cancer cell line HeLa and its molecular mechanism. Methods PCR technique was used to clone ISG15 and construct its recombinant plasmid and lentivirus vector, which was used to express ISG15. The effect of ISG15 on the proliferation of HeLa cells was detected by cell count method. The effect of ISG15 on the apoptosis of HeLa cells was detected by Western blot technique. The effect of ISG15 on the migration of HeLa cells was detected by cell scratch assay. Results the overexpression of ISG15 could inhibit the proliferation of HeLa cells. ISG15 could inhibit the migration of HeLa cells by inhibiting the apoptosis of HeLa cells induced by NF-B pathway. Conclusion: overexpression of ISG15 can induce apoptosis of HeLa cells by inhibiting NF-B pathway. Study on the role of ISG15 in the apoptosis of Hematological Disease cells induced by chloroiodoquine. Objective: to study the mechanism of ISG15 in anti-hematologic diseases. Methods RT-PCR and Western blot were used to detect the effect of chlorodoxyquine on the expression of mRNA and protein in hematological disease cells. (2) RT-PCR technique was used to detect the effect of chlorohydroxyquine on interferon signaling pathway. After overexpression of ISG15, The effects of overexpressed ISG15 on proliferation and apoptosis of hematological cells were detected by MTT assay and Western blot technique. Results Chloroiodoquine could significantly induce the expression of ISG15 in hematological cells. At the same time, it can induce the expression of debinding enzyme USP18 2) chloroiodoquine can activate the JAK-STAT signaling pathway and induce ISG15M3) overexpression of ISG15 can inhibit the proliferation of hematologic disease cells and promote its apoptosis. Conclusion: chloroiodoquine can induce up-regulation of ISG15 and overexpression of ISG15 can promote apoptosis of hematological cells.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R96

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