HNRPDL在肿瘤细胞中的功能研究

发布时间：2018-05-22 20:49

本文选题：HNRPDL + 结肠癌　；参考：《苏州大学》2016年硕士论文

【摘要】：目的:研究HNRPDL(Heterogeneous nuclear ribonucleoprotein D-like)在结肠癌及慢性髓细胞白血病(Chronic myeloid leukemia,CML)细胞中的功能。方法:(1)构建过表达HNRPDL和干扰其表达的逆转录病毒载体并制备病毒侵染SW620、K562等肿瘤细胞,Q-RT-PCR和western blot检测HNRPDL的表达情况;(2)干预HNRPDL表达后研究它对细胞增殖、迁移、周期、集落生成和致瘤能力的影响;(3)使用K562细胞研究HNRPDL对其伊马替尼(Imatinib mesylate,IM)反应性的影响;(4)在BaF3细胞中共表达BCR/ABL和HNRPDL,比较共表达与单独表达对细胞增殖和集落生成的影响。结果:(1)结肠癌样本中HNRPDL的表达明显高于对照样本。干扰HNRPDL的表达后,结肠癌细胞SW620的增殖、集落生成和迁移能力减弱,且在裸鼠皮下成瘤能力也降低(对照组:8/8;HNRPDL沉默组:5/8)。(2)细胞周期分析显示:干扰HNRPDL表达的SW620细胞与对照相比G0/G1期细胞增加,G2/M期细胞减少。(3)Western blot分析显示HNRPDL沉默抑制SW620细胞中cyclin D3的表达。(4)慢性髓细胞白血病患者骨髓中HNRPDL的表达较正常骨髓显著升高;沉默HNRPDL显著抑制CML细胞株(K562、MEG-01和BV173)和患者CD34+细胞的生长;也显著抑制了MEG-01细胞的裸鼠皮下成瘤能力(对照:6/8;shHNRPDL#1:1/8;shHNRPDL#2组:0/8)。(5)过表达HNRPDL促进BaF3细胞的生长及集落生成能力,并且能够减少BaF3细胞对mIL-3的依赖。HNRPDL也增强K562细胞的体外增殖。HNRPDL和BCR/ABL共表达细胞的生长较单独基因表达细胞显著增强。(6)HNRPDL沉默增加细胞对IM的敏感性,而过表达HNRPDL增加细胞对IM的耐受性。结论:(1)HNRPDL在结肠癌样本中异常高表达,干扰其表达后SW620细胞的体内外生长减缓,cyclin D3的表达减少、细胞周期阻滞在G0/G1期。(2)慢性髓细胞白血病患者骨髓中HNRPDL的表达较正常骨髓显著升高;HNRPDL沉默显著地抑制CML细胞的生长、集落生成和裸鼠皮下成瘤能力;过表达HNRPDL增强K562细胞增长,也增强BCR/ABL的促BaF3细胞生长能力;干预HNRPDL能调控CML细胞对IM的反应。
[Abstract]:Aim: to study the function of HNRPDL(Heterogeneous nuclear ribonucleoprotein D-like in Chronic myeloid leukemia (CML) cells of colon cancer and chronic myeloid leukemia. Methods the retrovirus vector expressing HNRPDL and interfering with its expression was constructed and infected with SW620- K562 and other tumor cells. Q-RT-PCR and western blot were used to detect the expression of HNRPDL. After interfering with the expression of HNRPDL, we studied the proliferation, migration and cycle of HNRPDL. K562 cell line was used to study the effect of HNRPDL on the reactivity of imatinib mesylatesil. BCR/ABL and HNRPDL were co-expressed in BaF3 cells, and the effects of co-expression and single expression on cell proliferation and colony formation were compared. Results the expression of HNRPDL in colon cancer samples was significantly higher than that in control samples. After interfering with the expression of HNRPDL, the proliferation, colony formation and migration of SW620 in colon cancer cells decreased. Cell cycle analysis showed that SW620 cells interfering with HNRPDL expression increased the number of G _ 2 / M phase cells in G0/G1 phase and decreased G _ 2 / M phase cells. Western blot analysis showed that HNRPDL silencing inhibited SW620 cells. The expression of cyclin D3 in bone marrow of patients with chronic myeloid leukemia was significantly higher than that of normal bone marrow. Silencing HNRPDL significantly inhibited the growth of CML cell lines K562MEG-01 and BV173, as well as the subcutaneous tumorigenesis of MEG-01 cells in nude mice (control group 6: 8 HNRPDL #1: 1 / 8 HNRPDL #2) over-expressed HNRPDL to promote the growth and colony formation of BaF3 cells. HNRPDL also enhanced the proliferation of K562 cells in vitro. HNRPDL and BCR/ABL co-expressed cells significantly increased the sensitivity of K562 cells to IM. Overexpression of HNRPDL increased cell tolerance to IM. Conclusion the expression of HNRPDL in human colon cancer samples is extremely high, and the expression of cyclin D3 decreases after interfering with the expression of HNRPDL in SW620 cells in vivo and in vitro. The expression of HNRPDL in the bone marrow of patients with chronic myeloid leukemia was significantly higher than that of normal bone marrow. HNRPDL silencing significantly inhibited the growth of CML cells, colony formation and subcutaneous tumorigenesis in nude mice, and overexpression of HNRPDL enhanced the growth of K562 cells. It also enhanced the ability of BCR/ABL to promote the growth of BaF3 cells, and interfered with HNRPDL to regulate the response of CML cells to IM.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R730.2

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