ILF2在肝细胞肝癌中的生物学功能及分子机制研究
发布时间：2018-07-27 12:23
【摘要】：背景在我国,肝细胞肝癌(Hepatocellular carcinoma,HCC)是一种高发的恶性肿瘤,90%的患者都有乙型肝炎的病史。中国每年死于肝癌约11万人,占全世界肝癌死亡人数的45%。肝癌患者初期症状并不明显,然而一旦出现症状,其病程大多已进入晚期,如果恶性程度高则其预后更差。虽然近几十年在肝癌切除、放射疗法、化学疗法及肝脏移植等治疗手段取得诸多进展,但是肝癌患者的长期预后仍然不理想。预后差的主要原因是肝癌的有效治疗靶点未找到以及肝癌发生发展的分子机制不明了。因此,寻找更具敏感性和特异性的肝癌的分子标志物和探究其在肝癌发生发展中的作用机制,将是进一步提高肝癌生存率的关键,对肝癌今后的早期诊断和早期治疗有着深远影响。目的本研究目的是探讨Interleukin enhancer binding factor 2(ILF2)在我院进行肝癌肝切除患者的肝癌组织和癌旁组织,以及肝癌细胞株中的表达水平,进而探索其与患者相关的各病理指数之间的联系,在TCGA数据库中分析其与肝癌患者预后的关系。用蛋白酶体抑制剂MG132分别在0,2,4,8,24h五个不同时间段处理肝癌细胞系,验证ILF2在肝癌组织及肝癌细胞中的高表达是通过转录水平引起。利用细胞学实验和体内成瘤技术,深度阐明ILF2在肝癌肝癌发生发展过程中所发挥的作用,以及其在肝癌形成中存在的信号通路。通过干扰ILF2的表达以及对肝癌细胞的培养基中添加OμM,5μM,1OμM,20μM不同浓度的人表皮生长因子受体(EGFR)的抑制剂厄洛替尼,探讨两种处理对细胞活性和增殖能力的协同作用。基于表达谱芯片的数据,对下游差异基因进行GO分析,利用KEGG数据库对差异基因进行Pathway分析,从而对ILF2下游分子或下游通路进行深一步分析和研究。方法1.提取27例HCC患者的肝癌肝切肿瘤组织及配对癌旁组织的mRNA,收集另外72例肝癌病人的肿瘤及癌旁石蜡组织,提取L02,Huh7,Bel-7402,MHCC-LM3,SMCC-7721和SK-HEP-1细胞系中的mRNA和8对肝癌组织及癌旁组织的蛋白质,利用Western免疫印迹、实时荧光定量PCR、免疫组化等技术验证白介素增强结合因子2(ILF2)在HCC肿瘤组织及各肝癌细胞系中的表达情况。2.根据肝癌组织的免疫组化结果,探究ILF2与肝癌病人临床病理参数之间的关系。3.利用TCGA数据库分析ILF2与肝癌患者生存率的关系。4.通过蛋白酶体抑制剂MG132处理肝癌细胞系,探索ILF2的表达升高是否通过转录水平的上调引起。5.通过转染ILF2小干扰RNA及慢病毒稳定转染过表达ILF2的方式,分别干扰或过表达ILF2,通过细胞学实验,如细胞克隆积聚实验、细胞凋亡实验和Cell Counting Kit-8(CCK-8)等,探索在肝癌细胞增殖和细胞凋亡的调控过程中ILF2所发挥的作用。6.为了探究ILF2在体内成瘤过程中的作用和影响,我们在裸鼠体内进行肿瘤细胞瘤体生成实验。7.运用Western blot技术寻找ILF2调控的下游分子,来探索ILF2在肝癌中发挥作用的分子机制。8.通过敲低ILF2的表达以及不同浓度的厄洛替尼处理肝癌细胞,运用Cell CountingKit-8(CCK-8)和克隆形成实验,验证不同处理后对肿瘤细胞活性和增殖能力的影响。9.基于表达谱芯片数据,对下游的显著性差异基因进行GO分析从而了解ILF2的生物学功能,从Biological Process,Cellular Component和Molecular Function 三个方面进行阐释,可以得到更加全面的信息。利用KEGG数据库对这些差异基因进行Pathway分析,从而有利于我们判断差异基因与哪些信号通路相关,全面了解ILF2的分子机制。结果1.在27对和另外72对肝癌病例中,我们发现,和正常癌旁组织相比,在肝癌组织中ILF2的RNA和蛋白质表达量明显升高。ILF2表达量与患者年龄、性别、TNM分期、肿瘤分级、血管侵袭和肝硬化没有相关性,与肿瘤大小具有密切关联性(p=0.043)。相对于正常肝永生化细胞L02细胞,其余5株细胞系的ILF2的表达升高。TCGA数据库对ILF2表达与肝癌患者预后进行分析,结果显示ILF2表达量升高与病人低生存率明显关联(p=0.0135)。2.在0,2,4,8,24h不同时间段用蛋白酶体抑制剂MG132处理肝癌细胞系,运用免疫印迹法检测不同时间处理细胞中的ILF2的表达。MG132处理后ILF2的表达没有明显变化。3.在体外功能实验中,我们发现ILF2过表达之后,可以促进肝癌细胞的克隆形成和增殖能力,而抑制ILF2的表达之后,肝癌细胞的克隆形成和增殖能力明显下降。4.流式凋亡实验结果发现,ILF2过表达之后,与对照组相比,肝癌细胞的凋亡比例减少,干扰ILF2后结果则相反。裸鼠皮下成瘤组织进行TUNEL实验,结果显示与对照组相比,过表达ILF2组的凋亡细胞明显减少。5.在裸鼠体内的瘤体生成实验中,Lenti-ILF2组得到的瘤体在重量和体积上均明显要高于对照组,免疫组化分析两组肿瘤组织的Ki67存在表达差异,并且过表达组的Ki67明显高表达。6.分子机制研究表明ILF2影响凋亡蛋白的表达,如BOK,BAX,BCL-2和cIAPl。ILF2升高导致的BCL-2和cIAP1的升高而BOK和BAX表达降低。而干扰ILF2后,下游蛋白的结果则相反。而对同为凋亡蛋白BAK来说,无论ILF2过表达或者干扰,BAK的表达均无变化。裸鼠皮下成瘤组织进行免疫组化实验,结果显示,与对照组相比,过表达ILF2组的BCL-2和cIAP1明显升高。7.与si-NC对照组相比,敲低ILF2的表达确实能增强厄洛替尼对肝癌细胞的活性抑制,两种处理同时进行能够发挥协同抑制作用。同样的浓度梯度处理细胞,在体外进行克隆实验,和CCK8结果一致的是,si-ILF2联合厄洛替尼处理明显能提高厄洛替尼对肝癌细胞的生长或增殖抑制作用。8.GO分析中,在Biological Process,ILF2与NF-KB、JNK和G1/S等信号通路密切相关;从Cellular Component发现,ILF2参与细胞外泌体、mRNA加工以及核蛋白复合物的组成;而通过Molecular Function的信息可知,其参与离子通道、RNA和DNA的结合。Pathway分析结果显示,脂肪酸代谢、糖代谢和AMPK信号通路与其密切相关。结论1.和肝癌邻近正常组织与正常肝细胞相比,在肝癌组织和肝癌细胞中,ILF2的mRNA和蛋白水平明显升高,肝癌患者中ILF2的高表达与肿瘤大小及低生存率密切相关,而且ILF2的表达上调主要是通过转录水平的调节。2.ILF2增强肝癌细胞的活性和增殖能力、抑制肿瘤细胞的凋亡,并且促进裸鼠瘤体的生长和增殖能力。3.在细胞水平上,BOK,BAX,BCL-2和cIAP1被证明由ILF2的调节,并且在体内成瘤肿瘤组织中证实ILF2过表达组出现BCL-2和cIAP1明显升高。4:体外克隆形成实验和细胞活性实验的研究结果均说明,si-ILF2联合不同浓度的EGFR抑制剂厄洛替尼在肝癌细胞的增殖能力和细胞活性抑制上有可能存在联合或协同作用。5.表达谱芯片结果显示,ILF2与mRNA加工、RNA/DNA相互结合、代谢过程以及AMPK信号通路等密切相关。
[Abstract]:Background in our country, Hepatocellular carcinoma (HCC) is a high incidence of malignant tumor. 90% of the patients have the history of hepatitis B. In China, about 110 thousand people died of liver cancer every year, and the initial symptoms of 45%. cancer patients in the death toll of all the world's liver cancer are not obvious. However, once the symptoms appear, the course of the disease is mostly in the late stage. If the malignancy is high, the prognosis is worse. Although many advances have been made in the past few decades in the treatment of liver cancer, radiotherapy, chemotherapy and liver transplantation, the long-term prognosis of the patients with liver cancer is still not ideal. The main reason for the poor prognosis is that the effective therapeutic targets for liver cancer have not been found and the molecular mechanism of the development of liver cancer. Therefore, the search for more sensitive and specific molecular markers of liver cancer and the mechanism of its role in the development of liver cancer will be the key to further improving the survival rate of HCC, and have a profound impact on the early diagnosis and early treatment of HCC. The purpose of this study is to explore the Interleukin enhancer binding f Actor 2 (ILF2) was used in the liver cancer tissues and para cancer tissues of the patients with hepatectomy and the expression level of the hepatoma cell lines in our hospital, and then the relationship between the pathological indexes related to the patients was explored, and the relationship with the prognosis of the patients with liver cancer was analyzed in the TCGA database. The proteasome inhibitor MG132 was in the 0,2,4,8,24h, respectively, in the 0,2,4,8,24h five. The high expression of ILF2 in liver cancer tissues and hepatoma cells is caused by the transcriptional level in different time periods. Using cytological experiments and in vivo tumorigenesis, the role of ILF2 in the development of hepatocellular carcinoma and the signaling pathway in the formation of liver cancer is deeply elucidated. By interfering with ILF2 The expression of O mu M, 5 mu M, 1O mu M, and 20 micron M of the human epidermal growth factor receptor (EGFR) inhibitor erlotinib were added to the hepatoma cells, and the synergistic effect of two treatments on the cell activity and proliferation ability was discussed. Based on the data of the expression chip, the downstream differential gene was analyzed by GO, and the KEGG database was used. The differential gene was analyzed by Pathway to carry out a deep analysis and Study on the downstream molecules or downstream pathways of the ILF2. Method 1. the mRNA of hepatoma liver resection and paracancerous tissue in 27 cases of HCC patients was extracted, and 72 other cancer patients and paraffin paraffin tissues were collected, and L02, Huh7, Bel-7402, MHCC-LM3, SMCC-7721 and SK-HEP- were extracted. The protein of mRNA and 8 in the 1 cell lines, using Western immunoblotting, real-time fluorescence quantitative PCR, immunohistochemistry and other techniques to verify the expression of interleukin enhanced binding factor 2 (ILF2) in HCC tumor tissue and all hepatoma cell lines.2. based on the immunohistochemical results of liver cancer tissue, ILF2 and liver cancer were explored. The relationship between human clinicopathological parameters.3. using TCGA database to analyze the relationship between ILF2 and the survival rate of patients with liver cancer.4. through the proteasome inhibitor MG132 treatment of liver cancer cell lines, explore whether the increase of ILF2 expression through the up regulation of transcriptional level causes.5. through the transfection of ILF2 small interference RNA and lentivirus stable transfection of ILF2. Interference or overexpression of ILF2, through cytological experiments, such as cell clone accumulation, apoptosis and Cell Counting Kit-8 (CCK-8), explore the role of ILF2 in the regulation of proliferation and apoptosis of hepatoma cells,.6. in order to explore the role and effect of ILF2 in the process of tumor formation in vivo, and we enter the body in nude mice. The tumor cell tumor formation experiment.7. uses Western blot technology to find the downstream molecules regulated by ILF2 to explore the molecular mechanism of ILF2 in the liver cancer,.8. by knocking low ILF2 expression and treating hepatoma cells with different concentrations of erlotinib, and using Cell CountingKit-8 (CCK-8) and clone formation experiments to verify the different treatment after treatment. The effect of.9. on the activity and proliferation of tumor cells is based on the expression spectrum chip data, and the GO analysis of the significant differentially differentially downstream genes is carried out to understand the biological function of ILF2, and the three aspects of Biological Process, Cellular Component and Molecular Function can be explained in order to get more comprehensive information. The use of KEGG database Pathway analysis of these differentially differentiable genes helps us to determine which signaling pathways are associated with the molecular mechanisms of ILF2. Results 1. in 27 pairs and 72 other cases of liver cancer, we found that the RNA and protein expression of ILF2 in the liver cancer tissues is significantly higher in.ILF2 expression than in normal para cancerous tissues. The volume was not associated with age, sex, TNM staging, tumor classification, vascular invasion and cirrhosis, and was closely related to the size of the tumor (p=0.043). Compared with normal liver immortalized L02 cells, the expression of ILF2 in the other 5 cell lines increased by.TCGA database to analyze the expression of ILF2 and the prognosis of liver cancer patients, and the results showed the ILF2 table. A significant association between the increase of amount and the low survival rate of the patient (p=0.0135).2. was used to treat the hepatocellular carcinoma cell lines with proteasome inhibitor MG132 at different time periods of 0,2,4,8,24h. The expression of ILF2 in the treated cells at different time was detected by Western blot, and the expression of ILF2 was not obviously changed after.MG132 treatment, and.3. in the function experiment in vitro, we found ILF2. After overexpression, it can promote the cloning and proliferation of hepatoma cells, and after inhibiting the expression of ILF2, the clone formation and proliferation ability of hepatoma cells decreased obviously by.4. flow apoptosis experiment. After ILF2 overexpression, the apoptosis ratio of hepatoma cells decreased compared with the control group, and the result of interference ILF2 was the opposite. Nude mice skin was the opposite. The results of TUNEL test in the lower tumor tissue showed that compared with the control group, the apoptotic cells overexpressed in the ILF2 group significantly reduced the tumor formation of.5. in the nude mice. The weight and volume of the Lenti-ILF2 group were significantly higher than those of the control group. The expression of Ki67 in the two groups of tumor tissues in the group of immuno histochemical analysis was different. The.6. molecular mechanism of the expression of Ki67 in the expression group indicates that ILF2 affects the expression of apoptotic proteins, such as BOK, BAX, BCL-2 and cIAPl.ILF2, which leads to the increase of BCL-2 and cIAP1, while BOK and BAX are reduced. The expression of BCL-2 and cIAP1 in the subcutaneous tissue of nude mice showed that compared with the control group, the expression of BCL-2 and cIAP1 in the overexpressed ILF2 group was significantly higher than that of the si-NC control group. The expression of low ILF2 could indeed enhance the inhibition of erlotinib on the activity of hepatoma cells, and the two treatments could play a synergistic inhibition at the same time. The same concentration gradient treatment cells were cloned in vitro, and the CCK8 results were consistent with that si-ILF2 combined with erlotinib significantly enhanced the.8.GO analysis of erlotinib's inhibitory effect on the growth or proliferation of hepatoma cells, which was closely related to Biological Process, ILF2 and NF-KB, JNK, and G1/S; from Cellular C. Omponent found that ILF2 participates in the extracellular secretory, mRNA processing and the composition of the nucleoprotein complex; and through the Molecular Function information, it is found that its involvement in ion channels, RNA and DNA binding.Pathway analysis results show that fatty acid metabolism, sugar metabolism and AMPK signaling pathway are closely related to it. Conclusion 1. and hepatocellular carcinoma are adjacent to normal tissues and positive cells. The mRNA and protein levels of ILF2 increased significantly in hepatoma and hepatoma cells. The high expression of ILF2 was closely related to the tumor size and low survival rate in HCC patients, and the up-regulated expression of ILF2 mainly enhanced the activity and proliferation of hepatoma cells through the regulation of.2.ILF2 at the transcriptional level and inhibited the tumor cells. Apoptosis, and promote the growth and proliferation of nude mice,.3. at the cell level, BOK, BAX, BCL-2 and cIAP1 have been proved to be regulated by ILF2, and in the tumor tissue of the body, it is confirmed that BCL-2 and cIAP1 in the ILF2 overexpression group are obviously elevated in the presence of.4: in vitro and in cell activity experiments. Different concentrations of EGFR inhibitor erlotinib may have joint or synergistic.5. expression profiles on the proliferation ability and cell activity inhibition of hepatoma cells. The results of ILF2 and mRNA processing, RNA/DNA binding, metabolic process and AMPK signaling pathway are closely related.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.7
本文编号：2147812
[Abstract]:Background in our country, Hepatocellular carcinoma (HCC) is a high incidence of malignant tumor. 90% of the patients have the history of hepatitis B. In China, about 110 thousand people died of liver cancer every year, and the initial symptoms of 45%. cancer patients in the death toll of all the world's liver cancer are not obvious. However, once the symptoms appear, the course of the disease is mostly in the late stage. If the malignancy is high, the prognosis is worse. Although many advances have been made in the past few decades in the treatment of liver cancer, radiotherapy, chemotherapy and liver transplantation, the long-term prognosis of the patients with liver cancer is still not ideal. The main reason for the poor prognosis is that the effective therapeutic targets for liver cancer have not been found and the molecular mechanism of the development of liver cancer. Therefore, the search for more sensitive and specific molecular markers of liver cancer and the mechanism of its role in the development of liver cancer will be the key to further improving the survival rate of HCC, and have a profound impact on the early diagnosis and early treatment of HCC. The purpose of this study is to explore the Interleukin enhancer binding f Actor 2 (ILF2) was used in the liver cancer tissues and para cancer tissues of the patients with hepatectomy and the expression level of the hepatoma cell lines in our hospital, and then the relationship between the pathological indexes related to the patients was explored, and the relationship with the prognosis of the patients with liver cancer was analyzed in the TCGA database. The proteasome inhibitor MG132 was in the 0,2,4,8,24h, respectively, in the 0,2,4,8,24h five. The high expression of ILF2 in liver cancer tissues and hepatoma cells is caused by the transcriptional level in different time periods. Using cytological experiments and in vivo tumorigenesis, the role of ILF2 in the development of hepatocellular carcinoma and the signaling pathway in the formation of liver cancer is deeply elucidated. By interfering with ILF2 The expression of O mu M, 5 mu M, 1O mu M, and 20 micron M of the human epidermal growth factor receptor (EGFR) inhibitor erlotinib were added to the hepatoma cells, and the synergistic effect of two treatments on the cell activity and proliferation ability was discussed. Based on the data of the expression chip, the downstream differential gene was analyzed by GO, and the KEGG database was used. The differential gene was analyzed by Pathway to carry out a deep analysis and Study on the downstream molecules or downstream pathways of the ILF2. Method 1. the mRNA of hepatoma liver resection and paracancerous tissue in 27 cases of HCC patients was extracted, and 72 other cancer patients and paraffin paraffin tissues were collected, and L02, Huh7, Bel-7402, MHCC-LM3, SMCC-7721 and SK-HEP- were extracted. The protein of mRNA and 8 in the 1 cell lines, using Western immunoblotting, real-time fluorescence quantitative PCR, immunohistochemistry and other techniques to verify the expression of interleukin enhanced binding factor 2 (ILF2) in HCC tumor tissue and all hepatoma cell lines.2. based on the immunohistochemical results of liver cancer tissue, ILF2 and liver cancer were explored. The relationship between human clinicopathological parameters.3. using TCGA database to analyze the relationship between ILF2 and the survival rate of patients with liver cancer.4. through the proteasome inhibitor MG132 treatment of liver cancer cell lines, explore whether the increase of ILF2 expression through the up regulation of transcriptional level causes.5. through the transfection of ILF2 small interference RNA and lentivirus stable transfection of ILF2. Interference or overexpression of ILF2, through cytological experiments, such as cell clone accumulation, apoptosis and Cell Counting Kit-8 (CCK-8), explore the role of ILF2 in the regulation of proliferation and apoptosis of hepatoma cells,.6. in order to explore the role and effect of ILF2 in the process of tumor formation in vivo, and we enter the body in nude mice. The tumor cell tumor formation experiment.7. uses Western blot technology to find the downstream molecules regulated by ILF2 to explore the molecular mechanism of ILF2 in the liver cancer,.8. by knocking low ILF2 expression and treating hepatoma cells with different concentrations of erlotinib, and using Cell CountingKit-8 (CCK-8) and clone formation experiments to verify the different treatment after treatment. The effect of.9. on the activity and proliferation of tumor cells is based on the expression spectrum chip data, and the GO analysis of the significant differentially differentially downstream genes is carried out to understand the biological function of ILF2, and the three aspects of Biological Process, Cellular Component and Molecular Function can be explained in order to get more comprehensive information. The use of KEGG database Pathway analysis of these differentially differentiable genes helps us to determine which signaling pathways are associated with the molecular mechanisms of ILF2. Results 1. in 27 pairs and 72 other cases of liver cancer, we found that the RNA and protein expression of ILF2 in the liver cancer tissues is significantly higher in.ILF2 expression than in normal para cancerous tissues. The volume was not associated with age, sex, TNM staging, tumor classification, vascular invasion and cirrhosis, and was closely related to the size of the tumor (p=0.043). Compared with normal liver immortalized L02 cells, the expression of ILF2 in the other 5 cell lines increased by.TCGA database to analyze the expression of ILF2 and the prognosis of liver cancer patients, and the results showed the ILF2 table. A significant association between the increase of amount and the low survival rate of the patient (p=0.0135).2. was used to treat the hepatocellular carcinoma cell lines with proteasome inhibitor MG132 at different time periods of 0,2,4,8,24h. The expression of ILF2 in the treated cells at different time was detected by Western blot, and the expression of ILF2 was not obviously changed after.MG132 treatment, and.3. in the function experiment in vitro, we found ILF2. After overexpression, it can promote the cloning and proliferation of hepatoma cells, and after inhibiting the expression of ILF2, the clone formation and proliferation ability of hepatoma cells decreased obviously by.4. flow apoptosis experiment. After ILF2 overexpression, the apoptosis ratio of hepatoma cells decreased compared with the control group, and the result of interference ILF2 was the opposite. Nude mice skin was the opposite. The results of TUNEL test in the lower tumor tissue showed that compared with the control group, the apoptotic cells overexpressed in the ILF2 group significantly reduced the tumor formation of.5. in the nude mice. The weight and volume of the Lenti-ILF2 group were significantly higher than those of the control group. The expression of Ki67 in the two groups of tumor tissues in the group of immuno histochemical analysis was different. The.6. molecular mechanism of the expression of Ki67 in the expression group indicates that ILF2 affects the expression of apoptotic proteins, such as BOK, BAX, BCL-2 and cIAPl.ILF2, which leads to the increase of BCL-2 and cIAP1, while BOK and BAX are reduced. The expression of BCL-2 and cIAP1 in the subcutaneous tissue of nude mice showed that compared with the control group, the expression of BCL-2 and cIAP1 in the overexpressed ILF2 group was significantly higher than that of the si-NC control group. The expression of low ILF2 could indeed enhance the inhibition of erlotinib on the activity of hepatoma cells, and the two treatments could play a synergistic inhibition at the same time. The same concentration gradient treatment cells were cloned in vitro, and the CCK8 results were consistent with that si-ILF2 combined with erlotinib significantly enhanced the.8.GO analysis of erlotinib's inhibitory effect on the growth or proliferation of hepatoma cells, which was closely related to Biological Process, ILF2 and NF-KB, JNK, and G1/S; from Cellular C. Omponent found that ILF2 participates in the extracellular secretory, mRNA processing and the composition of the nucleoprotein complex; and through the Molecular Function information, it is found that its involvement in ion channels, RNA and DNA binding.Pathway analysis results show that fatty acid metabolism, sugar metabolism and AMPK signaling pathway are closely related to it. Conclusion 1. and hepatocellular carcinoma are adjacent to normal tissues and positive cells. The mRNA and protein levels of ILF2 increased significantly in hepatoma and hepatoma cells. The high expression of ILF2 was closely related to the tumor size and low survival rate in HCC patients, and the up-regulated expression of ILF2 mainly enhanced the activity and proliferation of hepatoma cells through the regulation of.2.ILF2 at the transcriptional level and inhibited the tumor cells. Apoptosis, and promote the growth and proliferation of nude mice,.3. at the cell level, BOK, BAX, BCL-2 and cIAP1 have been proved to be regulated by ILF2, and in the tumor tissue of the body, it is confirmed that BCL-2 and cIAP1 in the ILF2 overexpression group are obviously elevated in the presence of.4: in vitro and in cell activity experiments. Different concentrations of EGFR inhibitor erlotinib may have joint or synergistic.5. expression profiles on the proliferation ability and cell activity inhibition of hepatoma cells. The results of ILF2 and mRNA processing, RNA/DNA binding, metabolic process and AMPK signaling pathway are closely related.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.7
【参考文献】
相关期刊论文 前1条
1 Mitsuro Kanda;Hiroyuki Sugimoto;Yasuhiro Kodera;;Genetic and epigenetic aspects of initiation and progression of hepatocellular carcinoma[J];World Journal of Gastroenterology;2015年37期
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