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  本文選題：HNRPDL + 結(jié)腸癌�。� 參考：《蘇州大學》2016年碩士論文

【摘要】：目的:研究HNRPDL(Heterogeneous nuclear ribonucleoprotein D-like)在結(jié)腸癌及慢性髓細胞白血病(Chronic myeloid leukemia,CML)細胞中的功能。方法:(1)構(gòu)建過表達HNRPDL和干擾其表達的逆轉(zhuǎn)錄病毒載體并制備病毒侵染SW620、K562等腫瘤細胞,Q-RT-PCR和western blot檢測HNRPDL的表達情況;(2)干預(yù)HNRPDL表達后研究它對細胞增殖、遷移、周期、集落生成和致瘤能力的影響;(3)使用K562細胞研究HNRPDL對其伊馬替尼(Imatinib mesylate,IM)反應(yīng)性的影響;(4)在BaF3細胞中共表達BCR/ABL和HNRPDL,比較共表達與單獨表達對細胞增殖和集落生成的影響。結(jié)果:(1)結(jié)腸癌樣本中HNRPDL的表達明顯高于對照樣本。干擾HNRPDL的表達后,結(jié)腸癌細胞SW620的增殖、集落生成和遷移能力減弱,且在裸鼠皮下成瘤能力也降低(對照組:8/8;HNRPDL沉默組:5/8)。(2)細胞周期分析顯示:干擾HNRPDL表達的SW620細胞與對照相比G0/G1期細胞增加,G2/M期細胞減少。(3)Western blot分析顯示HNRPDL沉默抑制SW620細胞中cyclin D3的表達。(4)慢性髓細胞白血病患者骨髓中HNRPDL的表達較正常骨髓顯著升高;沉默HNRPDL顯著抑制CML細胞株(K562、MEG-01和BV173)和患者CD34+細胞的生長;也顯著抑制了MEG-01細胞的裸鼠皮下成瘤能力(對照:6/8;shHNRPDL#1:1/8;shHNRPDL#2組:0/8)。(5)過表達HNRPDL促進BaF3細胞的生長及集落生成能力,并且能夠減少BaF3細胞對mIL-3的依賴。HNRPDL也增強K562細胞的體外增殖。HNRPDL和BCR/ABL共表達細胞的生長較單獨基因表達細胞顯著增強。(6)HNRPDL沉默增加細胞對IM的敏感性,而過表達HNRPDL增加細胞對IM的耐受性。結(jié)論:(1)HNRPDL在結(jié)腸癌樣本中異常高表達,干擾其表達后SW620細胞的體內(nèi)外生長減緩,cyclin D3的表達減少、細胞周期阻滯在G0/G1期。(2)慢性髓細胞白血病患者骨髓中HNRPDL的表達較正常骨髓顯著升高;HNRPDL沉默顯著地抑制CML細胞的生長、集落生成和裸鼠皮下成瘤能力;過表達HNRPDL增強K562細胞增長,也增強BCR/ABL的促BaF3細胞生長能力;干預(yù)HNRPDL能調(diào)控CML細胞對IM的反應(yīng)。
[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.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R730.2

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1 張鵬善;HNRPDL在腫瘤細胞中的功能研究[D];蘇州大學;2016年

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本文編號：1923555