类泛素化蛋白ISG15对恶性肿瘤细胞凋亡的诱导作用研究

发布时间：2018-03-06 04:35

  本文选题：宫颈癌细胞　切入点：ISG15　出处：《苏州大学》2014年硕士论文　论文类型：学位论文

【摘要】：第一章ISG15通过抑制NF-B途径诱导HeLa细胞的凋亡 目的：研究ISG15对宫颈癌细胞HeLa的促凋亡能力并探讨其分子机制。 方法：(1)使用PCR技术克隆ISG15并构建其重组质粒及慢病毒载体；（2）过表达ISG15后，采用细胞计数法检测ISG15对HeLa细胞增殖的影响；（3）采用Western blot技术检测ISG15对HeLa细胞促凋亡的影响；（4）采用细胞划痕实验检测ISG15对HeLa细胞迁移的影响。 结果：（1）过表达ISG15能够抑制HeLa细胞的增殖；(2)ISG15能够通过抑制NF-B途径诱导HeLa细胞的凋亡（；3）ISG15能够抑制HeLa细胞的迁移。 结论：过表达ISG15能够通过抑制NF-B途径诱导HeLa细胞的凋亡。 第二章ISG15在氯碘羟喹诱导的血液病细胞凋亡中的作用研究 目的：对ISG15在氯碘羟喹抗血液病作用中的机制研究。 方法：（1）采用RT-PCR和Western blot技术检测氯碘羟喹对血液病细胞中ISG15的mRNA和蛋白表达水平的影响；（2）采用RT-PCR技术检测氯碘羟喹对干扰素信号通路的影响；（3）采用慢病毒介导血液病细胞过表达ISG15之后，，通过MTT法和Western blot技术检测过表达的ISG15对血液病细胞的增殖和凋亡作用。 结果：（1）氯碘羟喹能够显著诱导血液病细胞中ISG15的表达，同时还能诱导其去结合酶USP18的表达；（2）氯碘羟喹能够激活JAK-STAT信号通路从而诱导ISG15；（3）过表达ISG15能够抑制血液病细胞的增殖并促进其凋亡。 结论：氯碘羟喹能够诱导ISG15的上调并且过表达ISG15能够促进血液病细胞的凋亡。
[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.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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