磷酸烯醇式丙酮酸羧激酶在肿瘤再生细胞代谢过程中作用的研究
发布时间：2018-08-25 07:38
【摘要】：第一部分上调表达PCK1对黑色素瘤再生细胞糖酵解侧枝反应的促进作用研究目的：肿瘤干细胞在肿瘤的发生、发展和治疗后复发过程中发挥重要作用。关于肿瘤细胞代谢的研究主要集中在混合细胞群体的层面,缺乏特异性针对肿瘤干细胞代谢特征及其分子调控机制的研究。本研究采用三维软纤维蛋白基质胶(3D fibrin)分离筛选的肿瘤干细胞(定义为肿瘤再生细胞,TRCs)为模型,探究糖异生的关键酶之一的胞浆型磷酸烯醇式丙酮酸羧激酶(PCK1)在黑色素瘤再生细胞中的表达及其发挥的代谢功能。方法：(1)以3D fibrin分离小鼠黑色素瘤B16细胞、肝癌H22细胞、淋巴瘤EL4细胞中的TRCs,然后使用RT-PCR、Real time PCR和Western blot检测这些TRCs和相应对照细胞以及小鼠未分化的间充质干细胞(mMSCs)和胚胎干细胞(mESCs)中PCK1的表达。(2)使用癌症基因数据库cbioportal中的基因表达数据分析PCK1与CD133、ALDH1A1和ABCG5等常用肿瘤干细胞标志在多细胞中的共表达关系；并采用Real time PCR检测PCK1在CD133+与CD133-B16细胞亚群中的表达差异。(3)免疫组化检测PCK1在9例人黑色素瘤临床标本中的表达；分离人原代黑色素瘤新鲜标本中的TRCs, real time PCR检测PCK 1的表达情况。(4) RT-PCR和Western blot检测B16TRCs中糖异生的另外两个关键酶FBP1和G6Pase的表达情况,以探讨PCK1是否介导糖异生的反应。(5)以siRNA沉默PCK1的表达后,观察B16 TRCs体外生长、葡萄糖消耗、乳酸释放以及中间代谢产物(如丝氨酸／甘氨酸,三磷酸甘油)等的变化,以探究PCK1发挥的代谢功能。(6)以siRNA沉默B16或H22 TRCs中PCK1的表达,观察其在动物体内成瘤能力。(7)以siRNA沉默B16 TRCs中PCK1的表达后,补充相关中间代谢产物,以观察对克隆生长的影响。(8)观察PCK1过表达对B16 TRCs体外生长的影响。(9)构建PCK1启动子控制的EGFP荧光表达B16细胞,分离出该细胞中的TRCs并置于普通硬质环境下连续培养,采用荧光显微镜动态观察荧光变化；将B16 TRCs置于普通硬质环境下连续培养,以RT-PCR、Real time PCR和Western blot检测PCK1的表达变化。(10)以小分子抑制剂或封闭抗体阻断B16 TRCs中相关信号通路位点后以Real time PCR检测PCK1的表达,初步探究PCK 1的表达调控机制。(11)亚硫酸氢盐测序法检测B16 TRCs中PCK1启动子区的甲基化水平,探讨DNA甲基化对PCK1表达调控的影响。结果：(1)B16细胞、H22细胞、EL4细胞TRCs相对于分化的肿瘤细胞上调表达PCK1,未分化的小鼠间充质干细胞和胚胎干细胞高表达PCK1。(2)PCK1与常用肿瘤干细胞标志CD133、ALDH1A1和ABCG5在多细胞中的表达呈正相关；PCK1在CD133+B16亚群细胞中的表达高于CD133-亚群细胞。(3)约1/3的黑色素瘤临床标本中检测到PCK1的高表达,分离自人原代黑色素瘤的TRCs上调表达PCK1。(4)B16 TRCs不表达糖异生的另外两个关键酶FBP1和G6Pase。(5) siRNA沉默PCK1表达后,B16 TRCs体外生长明显受抑,葡萄糖消耗和乳酸释放减少,甘氨酸和三磷酸甘油水平下降。(6) siRNA沉默PCK1表达后,B16和H22 TRCs的小鼠体内成瘤能力均减弱。(7)补充甘氨酸和(或)三磷酸甘油可部分逆转PCK1沉默后B16 TRCs受抑制的生长。(8)PCK1过表达可促进B16 TRCs的体外生长。(9)硬基质环境培养可诱导B16 TRCs中PCK1的下调表达。(10)抑制整合素αVβ3或P13K信号可诱导B16 TRCs中PCK1的表达下调,H3K9去甲基化可导致PCK1表达增加。(11)B16 TRCs和对照细胞中PCK1启动子均呈高度甲基化。结论：黑色素瘤再生细胞相对于分化的肿瘤细胞上调表达PCK1,因缺乏FBP1和G6Pase的表达,PCK1在肿瘤再生细胞中并不介导完整的糖异生,而是通过促进糖酵解的侧枝反应以增强肿瘤再生细胞中间代谢产物的合成。干扰PCK1的表达可以抑制黑色素瘤再生细胞的体外生长和体内成瘤能力。这些研究揭示PCK1上调表达是黑色素瘤再生细胞重要的代谢标志之一,有可能作为黑色素瘤治疗的一个新的靶点。第二部分PCK1、PCK2在乳腺癌肿瘤再生细胞中的表达研究目的：探究PCK1、PCK2在乳腺癌肿瘤再生细胞(TRCs)中的表达情况。方法：(1)采用三维软纤维蛋白基质胶(3D Fibrin)分离筛选MCF-7、T47D以及MDA-MB-231等人乳腺癌细胞株中的肿瘤再生细胞(TRCs),以RT-PCR检测PCK1在这些TRCs以及对照细胞中的基因表达。(2)以Western blot检测PCK1, PCK2在MCF-7、T47D、MDA-MB-231等来源TRCs以及相应的对照细胞中的表达变化。结果：(1)在MCF-7、T47D、MDA-MB-231等乳腺癌来源的TRCs和对照细胞中检测不到PCK1的基因表达。(2) MCF-7、T47D、MDA-MB-231等乳腺癌来源的TRCs相对于对照细胞上调表达PCK2。结论：乳腺癌再生细胞不表达PCK1而是上调表达其线粒体型的同工酶PCK2。PCK2上调表达可能是乳腺癌再生细胞重要的代谢特征之一,可为设计特异性针对乳腺癌肿瘤干细胞的抑制策略提供靶点。然而,关于PCK2的具体代谢功能还有待于进一步阐明。第三部分 PCK2偶联谷氨酰胺代谢途径调控乳腺癌肿瘤再生细胞的增殖研究目的：以乳腺癌MCF-7细胞为模型,探究丙酮酸羧化酶(PC)对PCK2上调表达的肿瘤再生细胞(TRCs)的生物学意义,并从葡萄糖和谷氨酰胺代谢偶联的角度对乳腺癌TRCs的细胞代谢特征进行新的诠释和探索。同时,我们尝试通过联合干扰PCK2和PC或谷氨酰胺裂解途径来抑制TRCs的肿瘤治疗新策略,为进一步研究肿瘤再生细胞的脂质代谢打下基础。方法：(1)使用三维软纤维蛋白基质胶筛选分离乳腺癌MCF-7细胞中的TRCs,以酶学方法和高效液相色谱分别检测MCF-7 TRCs和对照细胞葡萄糖和谷氨酰胺的利用效率,探究葡萄糖代谢与谷氨酰胺代谢的偶联。(2)观察无谷氨酰胺培养基培养的MCF-7 TRCs体外生长情况以研究TRCs对谷氨酰胺的依赖程度。(3)以Western blot检测丙酮酸羧化酶(PC)、谷氨酰胺转运蛋白(SLC1A5)、谷氨酰胺裂解酶(GLS)以及异柠檬酸脱氢酶2(IDH2)在MCF-7 TRCs和普通MCF-7细胞中的表达。(4)分别以siRNA联合干扰PCK2与PC、PCK2与GLS、PCK2与IDH2在MCF-7 TRCs中的表达,观察并统训MCF-7 TRCs克隆的体外生长情况。结果：(1)MCF-7 TRCs目对于普通MCF-7细胞消耗更多的葡萄糖和谷氨酰胺。(2)当剥夺了培养基中的谷氨酰胺,MCF-7 TRCs的克隆大小和数量急剧下降。(3)MCF-7 TRCs相对于对照细胞除上调表达PCK2外,还同时上调表达PC、SLC1A5和IDH2。(4)联合干扰PCK2与PC、PCK2与GLS或PCK2与IDH2较沉默单一位点对TRCs的抑制效果更明显。结论：在PCK2上调表达的乳腺癌MCF-7 TRCs中,为了补充线粒体内草酰乙酸的消耗以维持柠檬酸的合成,TRCs通过上调表达丙酮酸羧化酶以增加丙酮酸生成草酰乙酸的量。另外,TRCs还可通过增加谷氨酰胺的代谢来直接补充柠檬酸。通过PCK2上调可能介导的甘油骨架的合成增多以及谷氨酰胺裂解途径增强的脂肪酸代谢,TRCs可合成更多的脂类物质以满足其旺盛的代谢需求。联合靶向PCK2和PC或谷氨酰胺裂解途径可能是非常有潜在应用价值的针对肿瘤再生细胞的乳腺癌治疗新策略。
[Abstract]:Part 1 Upregulation of PCK1 expression in melanoma regenerating cells promotes glycolytic collateral reaction. Objective: Tumor stem cells play an important role in the occurrence, development and recurrence of melanoma. In this study, three-dimensional soft fibrin matrix glue (3D fibrin) was used to isolate and screen tumor stem cells (defined as tumor regeneration cells, TRCs) as a model to investigate the expression of cytoplasmic phosphoenolpyruvate carboxykinase 1 (PCK1), one of the key enzymes in glyconeogenesis, in melanoma regeneration cells. Methods: (1) TRCs were isolated from murine melanoma B16 cells, hepatoma H22 cells and lymphoma EL4 cells by 3D fibrin, and then detected by RT-PCR, Real-time PCR and Western blot for the expression of P in these TRCs and the corresponding control cells, as well as the undifferentiated mesenchymal stem cells (mMSCs) and embryonic stem cells (mESCs). CK1 expression. (2) The co-expression of PCK1 with CD133, ALDH1A1 and ABCG5 was analyzed by gene expression data from cbioportal, a cancer gene database. The expression of PCK1 in CD133 + and CD133-B16 cell subsets was detected by Real-time PCR. (3) PCK1 was detected by immunohistochemistry in 9 Black patients. The expression of the other two key enzymes FBP1 and G6Pase in B16TRCs was detected by RT-PCR and Western blot to investigate whether PCK1 mediated glyconeogenesis. (5) Silencing PCK1 with siRNA. After expression, the growth of B16 TRCs in vitro, glucose consumption, lactic acid release and intermediate metabolites (such as serine/glycine, glycerol triphosphate) were observed to explore the metabolic function of PCK1. (6) The expression of PCK1 in B16 or H22 TRCs was silenced by siRNA, and the tumorigenic ability of PCK1 in B16 TRCs was observed. (7) The expression of PCK1 in B16 TRCs was silenced by siRNA. (8) To observe the effect of overexpression of PCK1 on the growth of B16 TRCs in vitro. (9) To construct EGFP fluorescent expression B16 cells controlled by PCK1 promoter, TRCs were isolated from the cells and cultured continuously in normal hard environment. Fluorescence microscopy was used to dynamically observe the fluorescence. The expression of PCK1 was detected by RT-PCR, Real-time PCR and Western blot. (10) The expression of PCK1 was detected by Real-time PCR after blocking the signal pathway sites of B16 TRCs with small molecular inhibitors or blocking antibodies. Results: (1) The expression of PCK1 was up-regulated in B16 cells, H22 cells and EL4 cells compared with differentiated tumor cells, and the expression of PCK1 was up-regulated in undifferentiated mouse mesenchymal stem cells and embryonic stem cells. (2) PCK1 was over-expressed in undifferentiated mouse mesenchymal stem cells and embryonic stem cells. The expression of CD133, ALDH1A1 and ABCG5 was positively correlated in multicellular cells, and the expression of PCK1 in CD133+B16 subgroup was higher than that in CD133-subgroup. (3) The overexpression of PCK1 was detected in about one third of melanoma clinical specimens, and the expression of PCK1 was up-regulated in TRCs isolated from human primary melanoma. After the key enzymes FBP1 and G6Pase. (5) siRNA silenced the expression of PCK1, the growth of B16 TRCs was significantly inhibited, glucose consumption and lactic acid release were reduced, and the levels of glycine and triphosphate were decreased. (6) After the expression of PCK1 was silenced by siRNA, the tumorigenic ability of B16 and H22 TRCs in mice was weakened. (7) Glycine supplementation and/or triphosphate glycerol partially reversed P After CK1 silencing, the growth of B16 TRCs was inhibited. (8) Overexpression of PCK1 promoted the growth of B16 TRCs in vitro. (9) Hard substrate culture could induce the down-regulation of PCK1 expression in B16 TRCs. (10) Inhibition of integrin alpha V beta 3 or P13K signal could induce the down-regulation of PCK1 expression in B16 TRCs, and H3K9 demethylation could induce the increase of PCK1 expression in B16 TRCs and control cells. Conclusion: Compared with differentiated tumor cells, melanoma regenerated cells up-regulate the expression of PCK1. Due to the lack of FBP1 and G6Pase expression, PCK1 does not mediate complete Glyconeogenesis in tumor regenerated cells, but enhances the intermediate metabolites in tumor regenerated cells by promoting the lateral branching reaction of glycolysis. Synthesis. Interference with the expression of PCK1 inhibits the growth of melanoma regenerating cells in vitro and in vivo tumorigenicity. These studies suggest that the up-regulation of PCK1 is one of the important metabolic markers of melanoma regenerating cells and may be a new target for melanoma treatment. Part II PCK1, PCK2 in breast cancer regenerating cells Objective: To investigate the expression of PCK1 and PCK2 in tumor regeneration cells (TRCs) of breast cancer. Methods: (1) The tumor regeneration cells (TRCs) of human breast cancer cell lines MCF-7, T47D and MDA-MB-231 were isolated and screened by three-dimensional soft fibrin matrix glue (3D Fibrin), and detected by RT-PCR. The expression of PCK1 and PCK2 was detected by Western blot. Results: (1) The expression of PCK1 was not detected in the TRCs from MCF-7, T47D, MDA-MB-231 and the control cells. (2) The expression of PCK1 was not detected in the TRCs from MCF-7, T47D, MDA-MB-231 and other breast cancer sources. CONCLUSION: The up-regulation of PCK2 expression in breast cancer regeneration cells may be one of the important metabolic characteristics of breast cancer regeneration cells, and may provide a target for designing specific inhibitory strategies against breast cancer stem cells. The specific metabolic functions of PCK2-glutamine pathway in regulating the proliferation of breast cancer regeneration cells are to be further elucidated. Part III. Objective: To investigate the biological significance of pyruvate carboxylase (PC) on PCK2-up-regulated tumor regeneration cells (TRCs) using breast cancer MCF-7 cells as a model, and from glucose and glucose. Glutamine metabolic coupling provides a new interpretation and exploration of the cellular metabolic characteristics of TRCs in breast cancer. At the same time, we attempted to inhibit TRCs by interfering with PCK2 and PC or glutamine cleavage pathways in combination to lay a foundation for further study of lipid metabolism in tumor regenerated cells. TRCs isolated from breast cancer MCF-7 cells were screened by soft fibrin matrix glue. The utilization efficiency of glucose and glutamine in MCF-7 TRCs and control cells were detected by enzyme method and high performance liquid chromatography respectively. The coupling of glucose metabolism and glutamine metabolism was explored. (2) The growth of MCF-7 TRCs cultured in glutamine-free medium was observed in vitro. To study the dependence of TRCs on glutamine. (3) The expressions of pyruvate carboxylase (PC), glutamine transporter (SLC1A5), glutamine lyase (GLS) and isocitrate dehydrogenase 2 (IDH2) in MCF-7 TRCs and MCF-7 cells were detected by Western blot. (4) The expressions of PCK2 and PC, PCK2 and GLS, PCK2 and IDH2 were interfered with by siRNA, respectively. Results: (1) MCF-7 TRCs consumed more glucose and glutamine for normal MCF-7 cells. (2) When glutamine was deprived of the culture medium, the clonal size and number of MCF-7 TRCs decreased sharply. (3) MCF-7 TRCs increased the expression of glucose and glutamine in normal MCF-7 cells. In addition to PCK2, the expression of PC, SLC1A5 and IDH2 was up-regulated at the same time. (4) Combined interference of PCK2 with PC, PCK2 with GLS or PCK2 with IDH2 was more effective than silencing a single site on TRCs. CONCLUSION: In breast cancer MCF-7 TRCs up-regulated by PCK2, TRCs can maintain citric acid synthesis by supplementing the consumption of oxaloacetic acid in mitochondria. In addition, TRCs can directly supplement citric acid by increasing the metabolism of glutamine. By up-regulating the synthesis of Glycerol Skeleton and the metabolism of fatty acids by the cleavage pathway of glutamine, TRCs can synthesize more lipids to meet their needs. Strong metabolic demands. Targeting PCK2 and PC or glutamine cleavage pathways together may be a promising new strategy for breast cancer treatment targeting tumor regeneration cells.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R730.2
，
本文编号：2202206
[Abstract]:Part 1 Upregulation of PCK1 expression in melanoma regenerating cells promotes glycolytic collateral reaction. Objective: Tumor stem cells play an important role in the occurrence, development and recurrence of melanoma. In this study, three-dimensional soft fibrin matrix glue (3D fibrin) was used to isolate and screen tumor stem cells (defined as tumor regeneration cells, TRCs) as a model to investigate the expression of cytoplasmic phosphoenolpyruvate carboxykinase 1 (PCK1), one of the key enzymes in glyconeogenesis, in melanoma regeneration cells. Methods: (1) TRCs were isolated from murine melanoma B16 cells, hepatoma H22 cells and lymphoma EL4 cells by 3D fibrin, and then detected by RT-PCR, Real-time PCR and Western blot for the expression of P in these TRCs and the corresponding control cells, as well as the undifferentiated mesenchymal stem cells (mMSCs) and embryonic stem cells (mESCs). CK1 expression. (2) The co-expression of PCK1 with CD133, ALDH1A1 and ABCG5 was analyzed by gene expression data from cbioportal, a cancer gene database. The expression of PCK1 in CD133 + and CD133-B16 cell subsets was detected by Real-time PCR. (3) PCK1 was detected by immunohistochemistry in 9 Black patients. The expression of the other two key enzymes FBP1 and G6Pase in B16TRCs was detected by RT-PCR and Western blot to investigate whether PCK1 mediated glyconeogenesis. (5) Silencing PCK1 with siRNA. After expression, the growth of B16 TRCs in vitro, glucose consumption, lactic acid release and intermediate metabolites (such as serine/glycine, glycerol triphosphate) were observed to explore the metabolic function of PCK1. (6) The expression of PCK1 in B16 or H22 TRCs was silenced by siRNA, and the tumorigenic ability of PCK1 in B16 TRCs was observed. (7) The expression of PCK1 in B16 TRCs was silenced by siRNA. (8) To observe the effect of overexpression of PCK1 on the growth of B16 TRCs in vitro. (9) To construct EGFP fluorescent expression B16 cells controlled by PCK1 promoter, TRCs were isolated from the cells and cultured continuously in normal hard environment. Fluorescence microscopy was used to dynamically observe the fluorescence. The expression of PCK1 was detected by RT-PCR, Real-time PCR and Western blot. (10) The expression of PCK1 was detected by Real-time PCR after blocking the signal pathway sites of B16 TRCs with small molecular inhibitors or blocking antibodies. Results: (1) The expression of PCK1 was up-regulated in B16 cells, H22 cells and EL4 cells compared with differentiated tumor cells, and the expression of PCK1 was up-regulated in undifferentiated mouse mesenchymal stem cells and embryonic stem cells. (2) PCK1 was over-expressed in undifferentiated mouse mesenchymal stem cells and embryonic stem cells. The expression of CD133, ALDH1A1 and ABCG5 was positively correlated in multicellular cells, and the expression of PCK1 in CD133+B16 subgroup was higher than that in CD133-subgroup. (3) The overexpression of PCK1 was detected in about one third of melanoma clinical specimens, and the expression of PCK1 was up-regulated in TRCs isolated from human primary melanoma. After the key enzymes FBP1 and G6Pase. (5) siRNA silenced the expression of PCK1, the growth of B16 TRCs was significantly inhibited, glucose consumption and lactic acid release were reduced, and the levels of glycine and triphosphate were decreased. (6) After the expression of PCK1 was silenced by siRNA, the tumorigenic ability of B16 and H22 TRCs in mice was weakened. (7) Glycine supplementation and/or triphosphate glycerol partially reversed P After CK1 silencing, the growth of B16 TRCs was inhibited. (8) Overexpression of PCK1 promoted the growth of B16 TRCs in vitro. (9) Hard substrate culture could induce the down-regulation of PCK1 expression in B16 TRCs. (10) Inhibition of integrin alpha V beta 3 or P13K signal could induce the down-regulation of PCK1 expression in B16 TRCs, and H3K9 demethylation could induce the increase of PCK1 expression in B16 TRCs and control cells. Conclusion: Compared with differentiated tumor cells, melanoma regenerated cells up-regulate the expression of PCK1. Due to the lack of FBP1 and G6Pase expression, PCK1 does not mediate complete Glyconeogenesis in tumor regenerated cells, but enhances the intermediate metabolites in tumor regenerated cells by promoting the lateral branching reaction of glycolysis. Synthesis. Interference with the expression of PCK1 inhibits the growth of melanoma regenerating cells in vitro and in vivo tumorigenicity. These studies suggest that the up-regulation of PCK1 is one of the important metabolic markers of melanoma regenerating cells and may be a new target for melanoma treatment. Part II PCK1, PCK2 in breast cancer regenerating cells Objective: To investigate the expression of PCK1 and PCK2 in tumor regeneration cells (TRCs) of breast cancer. Methods: (1) The tumor regeneration cells (TRCs) of human breast cancer cell lines MCF-7, T47D and MDA-MB-231 were isolated and screened by three-dimensional soft fibrin matrix glue (3D Fibrin), and detected by RT-PCR. The expression of PCK1 and PCK2 was detected by Western blot. Results: (1) The expression of PCK1 was not detected in the TRCs from MCF-7, T47D, MDA-MB-231 and the control cells. (2) The expression of PCK1 was not detected in the TRCs from MCF-7, T47D, MDA-MB-231 and other breast cancer sources. CONCLUSION: The up-regulation of PCK2 expression in breast cancer regeneration cells may be one of the important metabolic characteristics of breast cancer regeneration cells, and may provide a target for designing specific inhibitory strategies against breast cancer stem cells. The specific metabolic functions of PCK2-glutamine pathway in regulating the proliferation of breast cancer regeneration cells are to be further elucidated. Part III. Objective: To investigate the biological significance of pyruvate carboxylase (PC) on PCK2-up-regulated tumor regeneration cells (TRCs) using breast cancer MCF-7 cells as a model, and from glucose and glucose. Glutamine metabolic coupling provides a new interpretation and exploration of the cellular metabolic characteristics of TRCs in breast cancer. At the same time, we attempted to inhibit TRCs by interfering with PCK2 and PC or glutamine cleavage pathways in combination to lay a foundation for further study of lipid metabolism in tumor regenerated cells. TRCs isolated from breast cancer MCF-7 cells were screened by soft fibrin matrix glue. The utilization efficiency of glucose and glutamine in MCF-7 TRCs and control cells were detected by enzyme method and high performance liquid chromatography respectively. The coupling of glucose metabolism and glutamine metabolism was explored. (2) The growth of MCF-7 TRCs cultured in glutamine-free medium was observed in vitro. To study the dependence of TRCs on glutamine. (3) The expressions of pyruvate carboxylase (PC), glutamine transporter (SLC1A5), glutamine lyase (GLS) and isocitrate dehydrogenase 2 (IDH2) in MCF-7 TRCs and MCF-7 cells were detected by Western blot. (4) The expressions of PCK2 and PC, PCK2 and GLS, PCK2 and IDH2 were interfered with by siRNA, respectively. Results: (1) MCF-7 TRCs consumed more glucose and glutamine for normal MCF-7 cells. (2) When glutamine was deprived of the culture medium, the clonal size and number of MCF-7 TRCs decreased sharply. (3) MCF-7 TRCs increased the expression of glucose and glutamine in normal MCF-7 cells. In addition to PCK2, the expression of PC, SLC1A5 and IDH2 was up-regulated at the same time. (4) Combined interference of PCK2 with PC, PCK2 with GLS or PCK2 with IDH2 was more effective than silencing a single site on TRCs. CONCLUSION: In breast cancer MCF-7 TRCs up-regulated by PCK2, TRCs can maintain citric acid synthesis by supplementing the consumption of oxaloacetic acid in mitochondria. In addition, TRCs can directly supplement citric acid by increasing the metabolism of glutamine. By up-regulating the synthesis of Glycerol Skeleton and the metabolism of fatty acids by the cleavage pathway of glutamine, TRCs can synthesize more lipids to meet their needs. Strong metabolic demands. Targeting PCK2 and PC or glutamine cleavage pathways together may be a promising new strategy for breast cancer treatment targeting tumor regeneration cells.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R730.2
，
本文编号：2202206
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