【摘要】：目的：腫瘤干細(xì)胞的存在讓腫瘤更具侵襲性、耐藥性以及轉(zhuǎn)移性。目前關(guān)于腫瘤代謝的研究集中在混合細(xì)胞群體的層面,缺乏針對腫瘤干細(xì)胞代謝特征以及其分子調(diào)控機制的研究。本研究采用三維軟蛋白基質(zhì)膠(3D Fibrin)培養(yǎng)獲取的腫瘤再生細(xì)胞(TRCs)為模型,探究線粒體型磷酸烯醇式丙酮酸(PCK2)的表達(dá)變化對腫瘤干細(xì)胞代謝的影響。方法：(1)RT-PCR和Western blot檢測比較小鼠黑色瘤細(xì)胞B16-F1、小鼠肝癌細(xì)胞H22、人喉癌細(xì)胞Hep-2以及人黑色素瘤細(xì)胞A875培養(yǎng)獲得的TRCs和相應(yīng)對照細(xì)胞中PCK2的表達(dá). RT-PCR和Western blot檢測比較小鼠胚胎干細(xì)胞(mESCs)在分化與未分化狀態(tài)下PCK2的表達(dá)。流式分選小鼠黑色素瘤細(xì)胞B1-F1中CD133+細(xì)胞亞群,RT-PCR檢測比較CD133+細(xì)胞亞群中PCK2的表達(dá)。收集兩例黑色素瘤臨床腫瘤病人標(biāo)本,分離腫瘤細(xì)胞培養(yǎng)獲取TRCs, RT-PCR檢測兩例病人標(biāo)本TRCs中PCK2的表達(dá)。(2)檢索TCGA數(shù)據(jù)庫并分析PCK2的表達(dá)與腫瘤病人生存期是否相關(guān)。(3)構(gòu)建PCK2過表達(dá)質(zhì)粒載體,轉(zhuǎn)染B16-F1TRCs,6小時后再將其尾靜脈接種小鼠,24天后觀察小鼠肺部成瘤情況；B16-F1細(xì)胞轉(zhuǎn)染PCK2 siRNA,6小時后再將其尾靜脈接種小鼠,24天后觀察小鼠肺部成瘤情況：構(gòu)建PCK2 Tet-On細(xì)胞株,將其種入3D fibrin膠中,在添加或不添加多西環(huán)素的情況下,比較克隆大小和數(shù)目,并比較TRCs對順鉑的敏感性。將PCK2 Tet-On細(xì)胞株,接種到小鼠皮下,分別用添加或不添加多西環(huán)素(Dox)的水喂養(yǎng),比較該細(xì)胞在小鼠皮下的成瘤能力。(4)分析比較B16-F1 TRCs和B16-F1葡萄糖消耗以及乳酸釋放量；過表達(dá)PCK2,比較B16-F1 TRCs葡萄糖消耗和乳酸釋放量；比較B16-F1 TRCs和對照細(xì)胞TCA循環(huán)中間物檸檬酸的含量；過表達(dá)PCK2,檢測B16-F1 TRCs檸檬酸的含量變化；siRNA沉默異檸檬酸脫氫酶IDH3A以及蘋果酸脫氫酶MDH2,觀察B16-F1 TRCs和對照細(xì)胞的生長情況并檢測其葡萄糖消耗；siRNA沉默檸檬酸轉(zhuǎn)運蛋白SLC25A1、檸檬酸裂解酶ACLY、脂肪酸合酶FASN以及蘋果酸酶ME1,檢測B16-F1 TRCs葡萄糖消耗；比較B16-F1 TRCs和對照細(xì)胞TCA循環(huán)代謝中間產(chǎn)物延胡索酸的含量；過表PCK2,檢測B16-F1 TRCs中延胡索酸含量變化；Western blot檢測B16-F1 TRCs和對照細(xì)胞中HIF1α和HIF2α的表達(dá)；調(diào)控PCK2表達(dá)后,Western blot檢測B16-F1 TRCs中HIF1α和HIF2α的表達(dá)；用質(zhì)譜檢測比較B16-F1 TRCs和對照細(xì)胞中DNA的5羥甲基胞嘧啶和5甲基胞嘧啶的含量：過表達(dá)PCK2,檢測B16-F1 TRCs DNA中5羥甲基胞嘧啶和5甲基胞嘧啶的含量變化；Seahorse XF24設(shè)備檢測比較B16-F1 TRCs和對照細(xì)胞的耗氧量水平；過表達(dá)PCK2后,檢測B16-F1 TRCs耗氧量變化：不同濃度的電子傳遞鏈復(fù)合物I抑制劑(Rotenone)、復(fù)合物V抑制劑寡霉素(Oligomycin)分別處理B16-F1 TRCs和對照細(xì)胞,用CCK-8試劑盒檢測比較細(xì)胞增殖情況,并檢測葡萄糖消耗量。結(jié)果：(1)B16-F1細(xì)胞、H22細(xì)胞、Hep-2細(xì)胞和A875細(xì)胞TRCs相對于普通腫瘤細(xì)胞,PCK2呈下調(diào)表達(dá)。mESC處于未分化狀態(tài)時低表達(dá)PCK2。PCK2在B16-F1 CD133+細(xì)胞亞群中的表達(dá)低于CD133細(xì)胞亞群。兩例黑色瘤病人標(biāo)本TRCs中PCK2呈低表達(dá)。(2)PCK2的表達(dá)與黑色素瘤、肺癌和胃癌病人的總生存期相關(guān)。(3)B16-F1 TRCs過表達(dá)PCK2后,在小鼠肺部形成轉(zhuǎn)移斑點數(shù)目減少；B16-F1轉(zhuǎn)染siRNA沉默PCK2后,在小鼠肺部形成轉(zhuǎn)移斑點數(shù)目增多；將PCK2 Tet-On穩(wěn)定細(xì)胞株種入3D fibrin中,在添加多西環(huán)素培養(yǎng)下,形成克隆的體積變小、數(shù)目減少,并且對順鉑敏感性變差。PCK2 Tet-On穩(wěn)定細(xì)胞株接種小鼠皮下,多西環(huán)素喂養(yǎng)組小鼠成瘤能力減弱。(4)與B16-F1相比,B16-F1TRCs葡萄糖消耗增多,乳酸釋放量增多；過表達(dá)PCK2后,B16-F1 TRCs葡萄糖消耗減少,乳酸產(chǎn)生量降低；B16-F1 TRCs檸檬酸的含量增高；過表達(dá)PCK2后,B16-F1 TRCs中檸檬酸的含量減少；B16-F1 TRCs沉默異檸檬酸脫氫酶IDH3A以及蘋果酸脫氫酶2后,克隆生長沒有明顯變化,葡萄糖消耗也沒有明顯差異,而B16-F1細(xì)胞生長明顯受抑制,葡萄糖消耗減少；沉默檸檬酸轉(zhuǎn)運蛋白SLC25A1、檸檬酸裂解酶ACLY、脂肪酸合酶FASN以及蘋果酸酶ME1,B16-F1 TRCs葡萄糖消耗明顯減少；B16-F1 TRCs中延胡索酸含量增多；過表達(dá)PCK2后,B16-F1TRCs中延胡索酸含量降低；在缺氧的情況下,B16-F1 TRCs中HIF蛋白的表達(dá)升高：過表達(dá)PCK2后,B16-F1 TRCs中HIF蛋白表達(dá)降低；B16-F1 TRCs細(xì)胞DNA的5-羥甲基化程度降低；過表達(dá)PCK2后,B16-F1 TRCs中DNA的5-羥甲基化程度增高；B16-F1 TRCs細(xì)胞耗氧量降低；過表達(dá)PCK2后,B16-F1 TRCs耗氧量增高：與B16-F1細(xì)胞相比,電子傳遞鏈抑制劑Rotenone和Oligomycin對B16-F1 TRCs細(xì)胞的抑制作用更差。結(jié)論：腫瘤再生細(xì)胞相對于對照細(xì)胞下調(diào)表達(dá)PCK2, PCK2低表達(dá)對維持腫瘤再生細(xì)胞的生長和成瘤至關(guān)重要,而且PCK2的表達(dá)與腫瘤病人的總生存期密切相關(guān)。過表達(dá)PCK2后,B16-F1 TRCs細(xì)胞體外生長受抑制,體內(nèi)致瘤能力減弱。B16-F1 TRCs糖酵解能力增強,氧化磷酸化水平降低。B16-F1 TRCs下調(diào)表達(dá)PCK2,發(fā)揮的生物學(xué)效應(yīng)的內(nèi)在機制可能是：①生成更多的檸檬酸,用于合成脂肪酸；②中間代謝物延胡索酸含量增高,進(jìn)而影響缺氧誘導(dǎo)因子的表達(dá),對細(xì)胞糖酵解產(chǎn)生影響；③中間代謝物延胡索酸含量增高,進(jìn)而影響DNA羥甲基化程度,維持細(xì)胞的未分化狀態(tài)；④降低細(xì)胞氧化磷酸化的水平,減少氧消耗。這些發(fā)現(xiàn)揭示PCK2的下調(diào)表達(dá)是黑色素腫瘤再生細(xì)胞重要的代謝標(biāo)志,在其代謝過程中發(fā)揮重要調(diào)節(jié)作用,有望作為黑色素瘤治療的靶標(biāo)。
[Abstract]:Objective: The presence of tumor stem cells has made the tumor more aggressive, resistant and metastatic. At present, the research on the tumor metabolism is focused on the level of the mixed cell population, and lacks the research on the metabolic characteristics of the tumor stem cells and the molecular control mechanism of the tumor stem cells. The effect of the expression of the mitochondrial phosphoenolpyruvate (PCK2) on the metabolism of the tumor stem cells was investigated by using the three-dimensional soft-protein matrix gel (3D Fibroin) culture to obtain the tumor-regeneration cells (TRCs). Methods: (1) The expression of PCK2 in mouse melanoma cells B16-F1, H22, Hep-2 and human melanoma cells A875 was compared with RT-PCR and Western blot. The expression of PCK2 in mouse embryonic stem cells (mESCs) was detected by RT-PCR and Western blot. The expression of PCK2 in CD133 + cell subpopulation was compared by RT-PCR. Two cases of melanoma clinical tumor were collected, and the expression of PCK2 in two patients with TRCs was detected by RT-PCR. (2) The TCGA database was searched and the expression of PCK2 was related to the survival of the tumor. and (3) constructing a PCK2 overexpression plasmid vector, transfecting B16-F1TRCs, and then inoculating the tail vein into a mouse after 6 hours, and observing the lung tumorigenicity of the mouse after 24 days; the B16-F1 cell is transfected with the PCK2 siRNA, and the tail vein is inoculated with the mouse after 6 hours, and the lung tumorigenicity of the mouse is observed after 24 days; and the PCK2 Tet-On cell line is constructed, In the case of adding or not adding doxycycline, the size and number of the clones were compared, and the sensitivity of the TRCs to cisplatin was compared. The PCK2 Tet-On cell line was inoculated to the subcutaneous of the mouse and was fed with water added or not added with doxycycline (Dox), respectively, and compared to the tumor-forming ability of the cell in the subcutaneous part of the mouse. (4) analyzing the consumption of B16-F1 TRCs and B16-F1 and the amount of lactic acid release; overexpressing PCK2, comparing the glucose consumption and the lactic acid release amount of B16-F1 TRCs; comparing the content of the citric acid in the B16-F1 TRCs and the control cell TCA; and detecting the content of the B16-F1 TRCs citric acid by the expression of the PCK2; The siRNA-silent isocitrate dehydrogenase IDH3A and the malic acid dehydrogenase MDH2 were used to observe the growth of B16-F1 TRCs and control cells and to detect their glucose consumption; siRNA-silent citric acid transport protein SLC25A1, citric acid lyase ACLY, fatty acid synthase FASN and malic enzyme ME1 were used to detect the glucose consumption of B16-F1 TRCs; The expression of HIF1 and HIF2 in B16-F1 TRCs and control cells was detected by Western blot, and the expression of HIF1 and HIF2 in B16-F1 TRCs was detected by Western blot. The content of 5-hydroxymethyl-1-and 5-methylpinacolin in B16-F1 TRCs and control cells was compared by mass-mass spectrometry. The content of 5-hydroxymethyl-1-and 5-methylpinacolin in B16-F1 TRCs and control cells was detected by mass-mass spectrometry. The level of oxygen consumption of B16-F1 TRCs and control cells was detected by Sehorse XF24 equipment. After the expression of PCK2, the oxygen consumption of B16-F1 TRCs was detected: the different concentrations of the electron transfer chain complex I inhibitor (Rotenone) and the compound V inhibitor, the oligonin, respectively treated B16-F1 TRCs and the control cells, and the proliferation of the cells was compared with the CCK-8 kit and the glucose consumption was detected. Results: (1) B16-F1 cells, H22 cells, Hep-2 cells and A875 cells TRCs were down-regulated with respect to normal tumor cells and PCK2. The expression of PCK2 and PCK2 in the subpopulation of B16-F1 CD133 + cells was lower than that of CD133 cells. In two cases of black tumor, PCK2 was expressed in low expression. (2) The expression of PCK2 is related to the overall survival of melanoma, lung cancer and gastric cancer. (3) After the B16-F1 TRCs overexpressing PCK2, the number of transfer spots in the lung of the mouse is reduced; after the B16-F1 transfected siRNA is silent PCK2, the number of transfer spots in the lung of the mouse is increased; the PCK2 Tet-On stable cell strain is seeded into the 3D fibrin, and the volume of the clone is reduced when the doxycycline is added, The number is reduced and the sensitivity to cisplatin is deteriorated. The tumor-forming ability of PCK2 Tet-On stable cell line was reduced under the subcutaneous and doxycycline-fed mice. (4) As compared with B16-F1, the consumption of B16-F1TRCs increased and the amount of lactic acid release increased; after the expression of PCK2, the glucose consumption of B16-F1 TRCs decreased and the amount of lactic acid production decreased; the content of the citric acid in B16-F1 TRCs was increased; after the expression of PCK2, the content of citric acid in B16-F1 TRCs decreased; When the B16-F1 TRCs were silent and the isocitrate dehydrogenase IDH3A and the malate dehydrogenase 2 were silent, the growth of the clones did not change significantly, and the consumption of glucose was not significantly different, and the B16-F1 cell growth was significantly inhibited, and the consumption of glucose decreased; the silent citric acid transport protein SLC25A1, the citric acid cleavage enzyme ACLY, In the case of hypoxia, the expression of HIF protein in B16-F1 TRCs increased: after overexpression of PCK2, the expression of HIF protein in B16-F1 TRCs increased: after overexpression of PCK2, The expression of HIF protein in B16-F1 TRCs decreased; the degree of 5-hydroxymethylation of the DNA of B16-F1 TRCs decreased; after the expression of PCK2, the degree of 5-hydroxymethylation of the DNA in B16-F1 TRCs increased; the oxygen consumption of B16-F1 TRCs decreased; after overexpression of PCK2, the oxygen consumption of B16-F1 TRCs increased: compared with B16-F1 cells, The inhibitory effect of the electron transfer chain inhibitor Rotenone and the oligonin on the B16-F1 TRCs cells is even worse. Conclusion: The low expression of PCK2 and PCK2 in tumor-regenerating cells is critical to the growth and tumorigenesis of tumor-regenerating cells, and the expression of PCK2 is closely related to the overall survival of tumor patients. After the expression of PCK2, the in vitro growth of B16-F1 TRCs cells was inhibited and the in vivo tumorigenicity decreased. The glycolysis ability of B16-F1 TRCs was enhanced and the level of oxidative phosphorylation was decreased. B16-F1 TRCs downregulated the expression of PCK2, and the intrinsic mechanism of the biological effects of B16-F1 may be: to produce more citric acid for the synthesis of fatty acids; to increase the content of the intermediate metabolite, to influence the expression of hypoxia-inducing factors, and to have an effect on the glycolysis of the cells; And the level of the oxidative phosphorylation of the cells is reduced, and the consumption of oxygen is reduced. These findings reveal that the down-regulation expression of PCK2 is an important metabolic marker in the regeneration of melanin tumor cells, plays an important role in the metabolism process, and is expected to be a target for melanoma treatment.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R739.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張玉勤;;細(xì)胞分化再活化治療腫瘤[J];國外醫(yī)學(xué)情報;1991年01期

2 David　Tenenbaum,饒新華;細(xì)胞之源[J];世界科學(xué);2000年11期

3 閆貴新;干細(xì)胞研究的重大進(jìn)展[J];畜牧獸醫(yī)科技信息;2000年11期

4 趙志力;付小兵;;正常與異常的細(xì)胞分化[J];感染.炎癥.修復(fù);2001年03期

5 趙志力;付小兵;;修復(fù)細(xì)胞分化的調(diào)控機制與鑒別[J];感染.炎癥.修復(fù);2001年04期

6 趙志力;修復(fù)細(xì)胞分化的調(diào)控機制與鑒別[J];中國危重病急救醫(yī)學(xué);2002年02期

7 趙志力;付小兵;;不同發(fā)育階段細(xì)胞的分化與調(diào)控機制[J];感染.炎癥.修復(fù);2002年01期

8 戴新貴;姚詠明;艾宇航;;輔助性T細(xì)胞17的研究進(jìn)展[J];生理科學(xué)進(jìn)展;2008年04期

9 ;細(xì)胞分化:一個兩階段的過程[J];中華婦幼臨床醫(yī)學(xué)雜志;2009年02期

10 黃紹勤;曹惠珍;;生命的基本單位——細(xì)胞[J];廣醫(yī)通訊;1980年03期

相關(guān)會議論文 前10條

1 徐志彥;;《細(xì)胞分化》教學(xué)案例(教學(xué)內(nèi)容安排1課時)[A];河北省教師教育學(xué)會第二屆中小學(xué)教師教學(xué)案例展論文集[C];2013年

2 尹青;張雷;李莉;謝丹丹;龔淼;;體外心肌樣環(huán)境對P19細(xì)胞向心肌細(xì)胞分化的誘導(dǎo)作用[A];中國解剖學(xué)會2012年年會論文文摘匯編[C];2012年

3 丁建東;;圖案化表面與干細(xì)胞分化[A];2013年全國高分子學(xué)術(shù)論文報告會論文摘要集——大會邀請報告[C];2013年

4 陳思;黃欣;陳少華;楊力建;鄭海濤;李揚秋;;BCL11B基因在T細(xì)胞分化和增殖中調(diào)節(jié)作用的研究[A];中國病理生理學(xué)會第九屆全國代表大會及學(xué)術(shù)會議論文摘要[C];2010年

5 江鍵;崔黎麗;宋誠榮;王小平;宋茂海;;負(fù)極性駐極體誘導(dǎo)3T3細(xì)胞表面電荷和游離鈣濃度的變化[A];第四屆中國功能材料及其應(yīng)用學(xué)術(shù)會議論文集[C];2001年

6 梁冀;于會梅;傅繼東;郭昂;楊黃恬;;內(nèi)質(zhì)網(wǎng)鈣離子釋放受體對干細(xì)胞分化的調(diào)控[A];中國病理生理學(xué)會受體、腫瘤和免疫專業(yè)委員會聯(lián)合學(xué)術(shù)會議論文匯編[C];2010年

7 羅家江;;TH17細(xì)胞的研究進(jìn)展[A];全國中西醫(yī)結(jié)合血液學(xué)學(xué)術(shù)會議論文匯編[C];2010年

8 項盈;金潔;沈丹;鄭強;謝純剛;賈冰冰;黃國平;王金福;;復(fù)蘇的人骨髓間充質(zhì)干細(xì)胞的分離、培養(yǎng)及向多系細(xì)胞分化的誘導(dǎo)[A];第10屆全國實驗血液學(xué)會議論文摘要匯編[C];2005年

9 汪洋;朱椰凡;余華軍;符竣惠;徐啟綱;曾其強;吳建波;張啟瑜;;三種全肝臟去細(xì)胞化流程對細(xì)胞外基質(zhì)的影響及細(xì)胞組分殘留量的研究[A];2013年浙江省外科學(xué)學(xué)術(shù)年會論文匯編[C];2013年

10 常平安;陳瑞;李薇;伍一軍;;神經(jīng)病靶標(biāo)酯酶的增強表達(dá)抑制哺乳動物細(xì)胞的增殖[A];中國環(huán)境誘變劑學(xué)會抗誘變劑和抗癌劑專業(yè)委員會第三次全國學(xué)術(shù)會議、中國毒理學(xué)會生化與分子毒理專業(yè)委員會第五屆學(xué)術(shù)交流會、貴州省環(huán)境誘變劑學(xué)會成立大會暨首屆學(xué)術(shù)交流會論文集[C];2004年

相關(guān)重要報紙文章 前10條

1 徐榮祥;人類生命細(xì)胞 再生物質(zhì)的發(fā)現(xiàn)與研究[N];健康報;2007年

2 張佳星;干細(xì)胞分化路徑存在差異性[N];中國礦業(yè)報;2008年

3 史軍;剝開黏合的細(xì)胞之書[N];健康報;2012年

4 常麗君;美生物學(xué)家打造出“細(xì)胞黑客”[N];科技日報;2010年

5 張佳星;分化之路 殊途同歸[N];科技日報;2008年

6 記者 施嘉奇　通訊員 王姿英;探索中藥對干細(xì)胞分化的作用[N];文匯報;2009年

7 記者陳超;日開發(fā)細(xì)胞間隔離培養(yǎng)技術(shù)[N];科技日報;2003年

8 陳超;科學(xué)家發(fā)現(xiàn)引發(fā)細(xì)胞分化的分子通道[N];科技日報;2010年

9 記者 白毅;我國科學(xué)家用iPS細(xì)胞首獲克隆豬[N];中國醫(yī)藥報;2013年

10 記者 錢錚;日本用人類iPS細(xì)胞首次制成血小板[N];新華每日電訊;2009年

相關(guān)博士學(xué)位論文 前10條

1 唐紹燦;IL-27、Th1及Th17細(xì)胞在MS中的表達(dá)及其作用機制研究[D];山東大學(xué);2015年

2 常凱凱;子宮內(nèi)膜異位灶微環(huán)境IL-10~+Th17細(xì)胞的形成及作用機制[D];復(fù)旦大學(xué);2014年

3 高華嵩;細(xì)胞重編程技術(shù)和移植治療視神經(jīng)損傷的實驗研究[D];復(fù)旦大學(xué);2014年

4 李艷明;Hsa-miR-200a調(diào)控紅細(xì)胞分化的功能及分子機制研究[D];中國科學(xué)院北京基因組研究所;2015年

5 張林;C-反應(yīng)蛋白通過對T細(xì)胞分化的直接調(diào)控參與后天免疫應(yīng)答[D];蘭州大學(xué);2015年

6 蔣云秋;樹突狀細(xì)胞Notch信號在1，，25-二羥維生素D3調(diào)節(jié)Th17分化中的作用研究[D];第三軍醫(yī)大學(xué);2015年

7 史莉瑾;急性腦出血患者外周血調(diào)節(jié)性T細(xì)胞與腦相關(guān)抗原的特征及其相關(guān)性研究[D];鄭州大學(xué);2015年

8 陳博;可注射細(xì)胞微球明膠復(fù)合物聯(lián)合血小板裂解液在牙組織工程中的應(yīng)用研究[D];第四軍醫(yī)大學(xué);2015年

9 章麗雅;血紅素加氧酶-1調(diào)抑Th17細(xì)胞在炎癥性腸病中的作用及機制研究[D];上海交通大學(xué);2013年

10 曹際森;miR-146a調(diào)控Treg細(xì)胞抑制小鼠心臟移植免疫排斥的研究[D];天津醫(yī)科大學(xué);2015年

相關(guān)碩士學(xué)位論文 前10條

1 丁佳敏;芪藍(lán)制劑對人舌鱗癌Tca8113細(xì)胞的抗增殖和促凋亡作用的機制研究[D];福建醫(yī)科大學(xué);2015年

2 張卓亞;間充質(zhì)干細(xì)胞對B6.MRL-Fas~(lpr)小鼠濾泡輔助T細(xì)胞的調(diào)節(jié)作用及機制研究[D];南京大學(xué);2015年

3 周明;As_2O_3聯(lián)合γ分泌酶抑制劑影響肝癌HepG_2細(xì)胞耐藥性的研究[D];石河子大學(xué);2015年

4 劉冰慧;凋亡細(xì)胞對巨噬細(xì)胞IL-12家族細(xì)胞因子調(diào)控的研究[D];河北醫(yī)科大學(xué);2015年

5 韓青青;Th22細(xì)胞及其效應(yīng)因子IL-22與類風(fēng)濕關(guān)節(jié)炎及合并間質(zhì)性肺病相關(guān)性的研究[D];河北醫(yī)科大學(xué);2015年

6 張騰飛;吉西他濱與AMD3100聯(lián)合應(yīng)用于膽管癌RBE細(xì)胞株對CXCR4/CXCL12軸的作用[D];河北醫(yī)科大學(xué);2015年

7 王們X ;EAAL細(xì)胞聯(lián)合全反式維甲酸對人肺腺癌細(xì)胞株A549體內(nèi)外作用的研究[D];河北醫(yī)科大學(xué);2015年

8 李超楠;兩種典型OPFRs對PC12細(xì)胞的神經(jīng)毒性及其機制探究[D];中國人民解放軍軍事醫(yī)學(xué)科學(xué)院;2015年

9 楊明;促腎上腺皮質(zhì)激素釋放因子1型受體經(jīng)cAMP/PKA信號通路對U87MG細(xì)胞凋亡機制的研究[D];河北醫(yī)科大學(xué);2015年

10 李廣康;RNAi沉默Rap2B基因?qū)盒赞D(zhuǎn)化BEAS-2B細(xì)胞的作用[D];鄭州大學(xué);2015年

本文編號：2486071