pp32/PHAPI调控肿瘤细胞迁移的作用研究
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摘要
目的意义:
磷蛋白32(phosphoprotein 32,pp32)是Malek等于1990年首次发现的,pp32是富含亮氨酸的酸性核蛋白(Leucine-rich acidic nuclear protein,LANP)家族成员之一。LANP家族成员包括pp32、PHAPI、Mapmodulin、I1PP2A、APRIL、HPPCn、pp32r1和pp32r2等。pp32由两个结构域组成,N端为5个亮氨酸组成的LRRs(Leucine-rich repeats)结构,是一个公认的蛋白间相互作用的构象。C端则是一个天冬氨酸和谷氨酸含量很高的酸性尾巴。伸展的C末端尾巴和三维构象为球形的N端LRRs区,整个蛋白结构类似一只蝌蚪。在C端的酸性区域中有一个典型的核定位信号NLS(nuclear localization signal)KRKR,是其行使核质穿梭行为的重要结构基础。
pp32是一个多功能的蛋白质,它涉及到很多生理过程,包括细胞增殖、凋亡、mRNA运输和转录。由于pp32已经被分离出过很多次,所以又被冠以不同的名称:pp32 (phosphoprotein 32)、Anp32a(acidic nuclear phosphoprotein32a)、PHAPI(putative HLA- associated protein I)、mapmodulin、Lanp(leucine-rich nuclear protein)和I1PP2A(inhibitor 1 protein phosphatas 2A)等。
目前对pp32的功能研究主要集中在以下几个方面:(1)能够抑制多种原癌基因(如ras、E1a、mutant p53和myc等)在大鼠成纤维细胞中的恶性转化。(2)促进人子宫颈癌细胞Hela和人乳腺癌细胞HMECs的凋亡。(3)抑制蛋白磷酸酶2A(PP2A)的活性。(4)作为mRNA结合蛋白HuR的配体,影响HuR蛋白的核质穿梭,参与mRNA的稳定性。(5)作为乙酰转移酶抑制剂(INHAT)的组成部分,参与调控组蛋白乙酰化和磷酸化过程,调控染色体结构。(6)参与细胞骨架动力学。(7)参与细胞内物质运输及信号传递。(8)参与组织发育。
一些研究表明,经低氧处理72h后,HepG2和Huh7细胞(它们与上皮细胞具有典型的相似性,并拥有发育良好的细胞连接)获得了梭形表型并普遍失去了细胞连接。为了进一步测定在低氧条件下HepG2和Huh7细胞的形状改变是否意味着EMT(上皮-间质转化),我们鉴定出相对于通常表型的HepG2细胞,过表达pp32的HepG2细胞形状发生改变。我们推测pp32会影响肿瘤细胞的运动及迁移能力。肿瘤转移是指恶性肿瘤细胞离开原来部位,通过血液循环或淋巴系统,到达其他部位并继续生长,形成新病灶的全过程。此新病灶称为转移灶,原来的肿瘤称原发灶。大多数肿瘤都具有转移性,这是临床上治愈肿瘤患者的主要障碍。
肿瘤细胞获得侵润、播散,并在转移部位存活下来这一复杂的过程受基因和细胞两种水平的调节。pp32与肿瘤转移之间的关系研究,很可能为以后的基因治疗提供有用的线索。本课题主要进行以下两方面的研究:1、pp32抑制肝癌细胞的迁移特性2、pp32影响Pc3M细胞的迁移特性
材料与方法:
利用western blots实验检测pp32在HepG2、SMMC7721、MHCC 97H、SK-hep1、MSC、和Ad-293等15种细胞中的表达。为了鉴定pp32在肝癌细胞迁移中的作用,我们将本实验室先前构建的HepG2-pcDNA3.1和HepG2-pcDNA3.1-pp32细胞株复苏,用800μg/ml的G418筛选三周后,再用400μg/ml的G418维持培养。Transwell小室实验检测细胞迁移能力,黏附测定实验检测细胞黏附能力。克隆形成率实验检测细胞增殖能力,用流式细胞仪检测细胞凋亡。并检测vinculin在HepG2、HepG2-pcDNA3.1和HepG2-pp32细胞中的表达。随后,利用尾静脉注射细胞实验在裸鼠体内初步检测了pp32对HepG2细胞迁移能力的影响。用裸鼠皮下移植瘤实验鉴定pp32在裸鼠体内对HepG2细胞增殖的影响。
另外,由于有研究发现,在前列腺癌中有pp32突变体(pp32r1和pp32r2)的存在,我们鉴定了pp32对高转移的前列腺癌细胞系Pc3M的影响。我们将本实验室先前保存的pcDNA3.1和pcDNA3.1-pp32菌液,摇菌,提取质粒后,将pcDNA3.1和pcDNA3.1-pp32质粒转染至Pc3M细胞,48小时后用700μg/ml的G418筛选三周,构建Pc3M-pcDNA3.1和Pc3M-pcDNA3.1-pp32稳定细胞株。Pc3M-pcDNA3.1和Pc3M-pcDNA3.1-pp32细胞用350μg/ml的G418维持培养。我们在体外鉴定了pp32对Pc3M细胞形状变化、迁移、黏附和增殖等特征的影响。
实验结果:
Western blots实验表明,pp32在15种细胞中均有表达。pp32过表达能够降低HepG2细胞的迁移和黏附能力。pp32能够抑制HepG2细胞中vinculin的表达。体内初步实验发现,pp32在裸鼠体内可能具有抑制HepG2细胞迁移的作用。相对于HepG2-pcDNA3.1细胞,HepG2-pp32细胞在裸鼠腋下所成的肿瘤大小无明显差异。
获得了稳定过表达pp32的Pc3M细胞株(Pc3M-pp32),pp32引起Pc3M细胞的形状改变,但不影响其恶性表型。pp32还引起人前列腺癌细胞Pc3M的迁移和黏附能力增加。pp32在HepG2和Pc3M细胞迁移中的作用是不同的,这非常有意思并为我们的研究带来了挑战。
结论与分析:
pp32是一个多功能的蛋白质,它不仅作为一个核蛋白发挥作用,而且有时还定位在细胞质中发挥作用。它不仅参与组织发育、信号转导、转录调控和其它多种生理过程,还能促进细胞凋亡、抑制肿瘤生长、抑制PP2A活性和具有其它功能。pp32能够抑制肝癌细胞HepG2的迁移特性,pp32能够促进Pc3M细胞的迁移。对pp32调控肿瘤细胞迁移的研究,不仅有利于肿瘤生物学的发展,而且有助于为人类肿瘤疾病寻找新的治疗途径。
Objects and significances:
Phosphoprotein 32 (pp32) was first discovered by Malek in 1990. Pp32 is a member of leucine-rich acidic nuclear proteins (Leucine-rich acidic nuclear protein, LANPs) family including PHAPI, Mapmodulin, I1PP2A, APRIL, HPPCn, pp32r1, pp32r2 and so on. Pp32 consists of two structual domains, and the N-terminal structure is composed of LRRs (Leucine-rich repeats) structure including five leucines, which is a recognized conformation of interaction between proteins. The C-terminal structure is an acid tail with high levels of aspartic acid and glutamic acid. The stretching C-terminal tail and the N-terminal LRRs zone whose three-dimensional conformation is spherical, and make the entire protein is similar to a tadpole. A typical nuclear localization signal NLS (nuclear localization signal) KRKR is in the C-terminal acidic region and it is an important structural foundation of shuttling between nucleus and cytoplasm.
Pp32, as a multifunctional protein, is involved in many physiological events which include cell proliferation, apoptosis, mRNA transport, and transcription. Pp32 has been isolated several times, and has therefore been assigned a variety of names: pp32 (phosphoprotein 32), Anp32a (acidic nuclear phosphoprotein32a), PHAPI (putative HLA-associated protein I), mapmodulin, Lanp (leucine-rich nuclear protein), and I1PP2A (inhibitor 1 protein phosphatase 2A).
The study of pp32 functions at present is focused on the following aspects:(1) Pp32 can inhibit the malignant transformation of a variety of proto-oncogenes in rat fibroblasts, including ras, E1a, mutant p53, myc and so on. (2) Pp32 promotes the apoptosis of human cervical carcinoma cells Hela and human breast cancer cells HMECs. (3) Pp32 inhibits the activity of PP2A. (4) As a ligand of mRNA binding protein HuR,pp32 affects the transport of pp32 from cell nucleus to cytoplasm. (5) As a constituent of INHAT (inhibitors of acetyltransferase), pp32 is involved in histone acetylation and phosphorylation and regulating the structure of chromosomes. (6) Pp32 is involved in cytoskeleton dynamics. (7) Pp32 is involved in substance transportion in cells and signal transduction. (8) It is involved in tissue development. Some research showed that 72 h after the hypoxia, HepG2 and Huh7 cells, which typically look like epithelia with well-developed cell junctions, acquired spindle shapes and generally lost cell contacts. To further test whether the morphologic changes in hypoxic HepG2 and Huh7 cells represent EMT (epithelial-mesenchymal transition), so the morphologic changes were detected in HepG2 cells overexpressing pp32 compared to wide type HepG2 cells. We speculated that pp32 affect tumor cell motility and migration. Tumor metastasis is defined as the whole process that malignant neoplasm cells shed and leave original sites, reach other parts through the blood or lymphatic system, continue to grow, and then form new lesions. The new lesion is named after metastatic focus, and the original tumor is called primary focus. The major tumors have the transferability which is the main obstacle to cure cancer patients clinically.
The complex process that a tumor cell obtains abilities of invasion and spread and survives at metastatic sites is regulated by genic and cellular levels. Studies of relations between pp32 and tumor metastasis probably provide useful clues for further gene therapy. This object majors in studying the following two aspects: 1、Pp32 inhibits hepatocellular carcinoma cell migration characteristics. 2、Pp32 infects Pc3M cell migration characteristics.
Materials and methods:
The protein level of pp32 was detected by western blots experiments in 15 cell lines including HepG2, SMMC7721, MHCC 97H, SK-hep1, MSC, Ad-293, and so on. In order to identify roles of pp32 in hepatoma carcinoma cells migration, we resuscitated HepG2- pcDNA3.1 cells and HepG2-pcDNA3.1-pp32 cells previously constructed, we sifted cells with 800μg/ml G418 for three weeks, and then we sustaining cultured cells with 400μg/ml G418. The ability of cell migration was detected by transwell assays, and the ability of cell adhesion was detected by adhesion experiments. The ability of cell proliferation was determined by clongenic assays, and cell apoptosis was determined by flow cytometries. The expression of vinculin in HepG2, HepG2-pcDNA3.1 and HepG2-pp32 were detected. Whereafter, the impact of pp32 on the metastasis of HepG2 cells of nude mice in vivo was primarily examined by experiments of injecting cells via tail vein. The impact of pp32 on the proliferation of HepG2 cells of nude mice in vivo was examined by tumor xenograft experiments.
In addition, some studies have found that there are pp32 mutants in prostate such as pp32r1 and pp32r2, so we identified the impact of pp32 on the highly metastatic prostate cancer cell lines Pc3M. We shaked the bacilli of pcDNA3.1 and pcDNA3.1-pp32 previously saved, after the extraction of plasmids, plasmid pcDNA3.1 and plasmid pcDNA3.1-pp32 were transfected into Pc3M cells, after 48 hours we sifted cells with 700μg/ml G418 for three weeks in order to construct stable Pc3M-pcDNA3.1 and Pc3M-pcDNA3.1-pp32 cell lines. Pc3M-pcDNA3.1 and Pc3M-pcDNA3.1-pp32 cells were cultivated with 350μg/ml G418. We have identified the impact of pp32 on Pc3M cells traits in aspects of morphologic changes, migration, adhesion and proliferation in vitro.
Results:
Western blots showed that the expression of pp32 was detected in all of 15 cell lines. Pp32 over-expression reduced the migration and adhesion abilities of HepG2 cells. Pp32 inhibited the expression of vinculin in HepG2 cells. In vivo primary experimental results showed that pp32 might inhibit HepG2 cells metastasis in nude mice. Compared to HepG2-pcDNA3.1 cells, the tumor size formed by HepG2-pp32 cells in the nude mice armpit had no significant differences.
Pc3M cell lines stable over-expressing pp32 (Pc3M-pp32) were obtained, pp32 could lead to shape changes of Pc3M cells, but did not affect their malignant phenotypes. Pp32 also led to promotion of the migration and adhesion of human prostate cancer cells Pc3M. The role of pp32 in cells migration of HepG2 and Pc3M cells is different, which was very interesting and gives us challenges.
Conclusions and analysis:
Pp32 is a multifunctional protein, which not only plays a role as a nuclear protein, but sometimes pp32 is located in the cytoplasm to take action. It is not only involved in tissue development, signal transduction, transcriptional regulation and other physiological processes, but also can promote cell apoptosis, inhibit tumor growth, inhibit the activity of PP2A and have other functions. Our studies showed that pp32 could inhibit cell migration characteristics of hepatoma carcinoma cells lines HepG2, but pp32 could stimulate Pc3M cell migration. Studies of pp32 regulating cell migration are not only conducive to the development of tumor biology, but also contribute to find new pathways to the treatment of human cancer.
磷蛋白32(phosphoprotein 32,pp32)是Malek等于1990年首次发现的,pp32是富含亮氨酸的酸性核蛋白(Leucine-rich acidic nuclear protein,LANP)家族成员之一。LANP家族成员包括pp32、PHAPI、Mapmodulin、I1PP2A、APRIL、HPPCn、pp32r1和pp32r2等。pp32由两个结构域组成,N端为5个亮氨酸组成的LRRs(Leucine-rich repeats)结构,是一个公认的蛋白间相互作用的构象。C端则是一个天冬氨酸和谷氨酸含量很高的酸性尾巴。伸展的C末端尾巴和三维构象为球形的N端LRRs区,整个蛋白结构类似一只蝌蚪。在C端的酸性区域中有一个典型的核定位信号NLS(nuclear localization signal)KRKR,是其行使核质穿梭行为的重要结构基础。
pp32是一个多功能的蛋白质,它涉及到很多生理过程,包括细胞增殖、凋亡、mRNA运输和转录。由于pp32已经被分离出过很多次,所以又被冠以不同的名称:pp32 (phosphoprotein 32)、Anp32a(acidic nuclear phosphoprotein32a)、PHAPI(putative HLA- associated protein I)、mapmodulin、Lanp(leucine-rich nuclear protein)和I1PP2A(inhibitor 1 protein phosphatas 2A)等。
目前对pp32的功能研究主要集中在以下几个方面:(1)能够抑制多种原癌基因(如ras、E1a、mutant p53和myc等)在大鼠成纤维细胞中的恶性转化。(2)促进人子宫颈癌细胞Hela和人乳腺癌细胞HMECs的凋亡。(3)抑制蛋白磷酸酶2A(PP2A)的活性。(4)作为mRNA结合蛋白HuR的配体,影响HuR蛋白的核质穿梭,参与mRNA的稳定性。(5)作为乙酰转移酶抑制剂(INHAT)的组成部分,参与调控组蛋白乙酰化和磷酸化过程,调控染色体结构。(6)参与细胞骨架动力学。(7)参与细胞内物质运输及信号传递。(8)参与组织发育。
一些研究表明,经低氧处理72h后,HepG2和Huh7细胞(它们与上皮细胞具有典型的相似性,并拥有发育良好的细胞连接)获得了梭形表型并普遍失去了细胞连接。为了进一步测定在低氧条件下HepG2和Huh7细胞的形状改变是否意味着EMT(上皮-间质转化),我们鉴定出相对于通常表型的HepG2细胞,过表达pp32的HepG2细胞形状发生改变。我们推测pp32会影响肿瘤细胞的运动及迁移能力。肿瘤转移是指恶性肿瘤细胞离开原来部位,通过血液循环或淋巴系统,到达其他部位并继续生长,形成新病灶的全过程。此新病灶称为转移灶,原来的肿瘤称原发灶。大多数肿瘤都具有转移性,这是临床上治愈肿瘤患者的主要障碍。
肿瘤细胞获得侵润、播散,并在转移部位存活下来这一复杂的过程受基因和细胞两种水平的调节。pp32与肿瘤转移之间的关系研究,很可能为以后的基因治疗提供有用的线索。本课题主要进行以下两方面的研究:1、pp32抑制肝癌细胞的迁移特性2、pp32影响Pc3M细胞的迁移特性
材料与方法:
利用western blots实验检测pp32在HepG2、SMMC7721、MHCC 97H、SK-hep1、MSC、和Ad-293等15种细胞中的表达。为了鉴定pp32在肝癌细胞迁移中的作用,我们将本实验室先前构建的HepG2-pcDNA3.1和HepG2-pcDNA3.1-pp32细胞株复苏,用800μg/ml的G418筛选三周后,再用400μg/ml的G418维持培养。Transwell小室实验检测细胞迁移能力,黏附测定实验检测细胞黏附能力。克隆形成率实验检测细胞增殖能力,用流式细胞仪检测细胞凋亡。并检测vinculin在HepG2、HepG2-pcDNA3.1和HepG2-pp32细胞中的表达。随后,利用尾静脉注射细胞实验在裸鼠体内初步检测了pp32对HepG2细胞迁移能力的影响。用裸鼠皮下移植瘤实验鉴定pp32在裸鼠体内对HepG2细胞增殖的影响。
另外,由于有研究发现,在前列腺癌中有pp32突变体(pp32r1和pp32r2)的存在,我们鉴定了pp32对高转移的前列腺癌细胞系Pc3M的影响。我们将本实验室先前保存的pcDNA3.1和pcDNA3.1-pp32菌液,摇菌,提取质粒后,将pcDNA3.1和pcDNA3.1-pp32质粒转染至Pc3M细胞,48小时后用700μg/ml的G418筛选三周,构建Pc3M-pcDNA3.1和Pc3M-pcDNA3.1-pp32稳定细胞株。Pc3M-pcDNA3.1和Pc3M-pcDNA3.1-pp32细胞用350μg/ml的G418维持培养。我们在体外鉴定了pp32对Pc3M细胞形状变化、迁移、黏附和增殖等特征的影响。
实验结果:
Western blots实验表明,pp32在15种细胞中均有表达。pp32过表达能够降低HepG2细胞的迁移和黏附能力。pp32能够抑制HepG2细胞中vinculin的表达。体内初步实验发现,pp32在裸鼠体内可能具有抑制HepG2细胞迁移的作用。相对于HepG2-pcDNA3.1细胞,HepG2-pp32细胞在裸鼠腋下所成的肿瘤大小无明显差异。
获得了稳定过表达pp32的Pc3M细胞株(Pc3M-pp32),pp32引起Pc3M细胞的形状改变,但不影响其恶性表型。pp32还引起人前列腺癌细胞Pc3M的迁移和黏附能力增加。pp32在HepG2和Pc3M细胞迁移中的作用是不同的,这非常有意思并为我们的研究带来了挑战。
结论与分析:
pp32是一个多功能的蛋白质,它不仅作为一个核蛋白发挥作用,而且有时还定位在细胞质中发挥作用。它不仅参与组织发育、信号转导、转录调控和其它多种生理过程,还能促进细胞凋亡、抑制肿瘤生长、抑制PP2A活性和具有其它功能。pp32能够抑制肝癌细胞HepG2的迁移特性,pp32能够促进Pc3M细胞的迁移。对pp32调控肿瘤细胞迁移的研究,不仅有利于肿瘤生物学的发展,而且有助于为人类肿瘤疾病寻找新的治疗途径。
Objects and significances:
Phosphoprotein 32 (pp32) was first discovered by Malek in 1990. Pp32 is a member of leucine-rich acidic nuclear proteins (Leucine-rich acidic nuclear protein, LANPs) family including PHAPI, Mapmodulin, I1PP2A, APRIL, HPPCn, pp32r1, pp32r2 and so on. Pp32 consists of two structual domains, and the N-terminal structure is composed of LRRs (Leucine-rich repeats) structure including five leucines, which is a recognized conformation of interaction between proteins. The C-terminal structure is an acid tail with high levels of aspartic acid and glutamic acid. The stretching C-terminal tail and the N-terminal LRRs zone whose three-dimensional conformation is spherical, and make the entire protein is similar to a tadpole. A typical nuclear localization signal NLS (nuclear localization signal) KRKR is in the C-terminal acidic region and it is an important structural foundation of shuttling between nucleus and cytoplasm.
Pp32, as a multifunctional protein, is involved in many physiological events which include cell proliferation, apoptosis, mRNA transport, and transcription. Pp32 has been isolated several times, and has therefore been assigned a variety of names: pp32 (phosphoprotein 32), Anp32a (acidic nuclear phosphoprotein32a), PHAPI (putative HLA-associated protein I), mapmodulin, Lanp (leucine-rich nuclear protein), and I1PP2A (inhibitor 1 protein phosphatase 2A).
The study of pp32 functions at present is focused on the following aspects:(1) Pp32 can inhibit the malignant transformation of a variety of proto-oncogenes in rat fibroblasts, including ras, E1a, mutant p53, myc and so on. (2) Pp32 promotes the apoptosis of human cervical carcinoma cells Hela and human breast cancer cells HMECs. (3) Pp32 inhibits the activity of PP2A. (4) As a ligand of mRNA binding protein HuR,pp32 affects the transport of pp32 from cell nucleus to cytoplasm. (5) As a constituent of INHAT (inhibitors of acetyltransferase), pp32 is involved in histone acetylation and phosphorylation and regulating the structure of chromosomes. (6) Pp32 is involved in cytoskeleton dynamics. (7) Pp32 is involved in substance transportion in cells and signal transduction. (8) It is involved in tissue development. Some research showed that 72 h after the hypoxia, HepG2 and Huh7 cells, which typically look like epithelia with well-developed cell junctions, acquired spindle shapes and generally lost cell contacts. To further test whether the morphologic changes in hypoxic HepG2 and Huh7 cells represent EMT (epithelial-mesenchymal transition), so the morphologic changes were detected in HepG2 cells overexpressing pp32 compared to wide type HepG2 cells. We speculated that pp32 affect tumor cell motility and migration. Tumor metastasis is defined as the whole process that malignant neoplasm cells shed and leave original sites, reach other parts through the blood or lymphatic system, continue to grow, and then form new lesions. The new lesion is named after metastatic focus, and the original tumor is called primary focus. The major tumors have the transferability which is the main obstacle to cure cancer patients clinically.
The complex process that a tumor cell obtains abilities of invasion and spread and survives at metastatic sites is regulated by genic and cellular levels. Studies of relations between pp32 and tumor metastasis probably provide useful clues for further gene therapy. This object majors in studying the following two aspects: 1、Pp32 inhibits hepatocellular carcinoma cell migration characteristics. 2、Pp32 infects Pc3M cell migration characteristics.
Materials and methods:
The protein level of pp32 was detected by western blots experiments in 15 cell lines including HepG2, SMMC7721, MHCC 97H, SK-hep1, MSC, Ad-293, and so on. In order to identify roles of pp32 in hepatoma carcinoma cells migration, we resuscitated HepG2- pcDNA3.1 cells and HepG2-pcDNA3.1-pp32 cells previously constructed, we sifted cells with 800μg/ml G418 for three weeks, and then we sustaining cultured cells with 400μg/ml G418. The ability of cell migration was detected by transwell assays, and the ability of cell adhesion was detected by adhesion experiments. The ability of cell proliferation was determined by clongenic assays, and cell apoptosis was determined by flow cytometries. The expression of vinculin in HepG2, HepG2-pcDNA3.1 and HepG2-pp32 were detected. Whereafter, the impact of pp32 on the metastasis of HepG2 cells of nude mice in vivo was primarily examined by experiments of injecting cells via tail vein. The impact of pp32 on the proliferation of HepG2 cells of nude mice in vivo was examined by tumor xenograft experiments.
In addition, some studies have found that there are pp32 mutants in prostate such as pp32r1 and pp32r2, so we identified the impact of pp32 on the highly metastatic prostate cancer cell lines Pc3M. We shaked the bacilli of pcDNA3.1 and pcDNA3.1-pp32 previously saved, after the extraction of plasmids, plasmid pcDNA3.1 and plasmid pcDNA3.1-pp32 were transfected into Pc3M cells, after 48 hours we sifted cells with 700μg/ml G418 for three weeks in order to construct stable Pc3M-pcDNA3.1 and Pc3M-pcDNA3.1-pp32 cell lines. Pc3M-pcDNA3.1 and Pc3M-pcDNA3.1-pp32 cells were cultivated with 350μg/ml G418. We have identified the impact of pp32 on Pc3M cells traits in aspects of morphologic changes, migration, adhesion and proliferation in vitro.
Results:
Western blots showed that the expression of pp32 was detected in all of 15 cell lines. Pp32 over-expression reduced the migration and adhesion abilities of HepG2 cells. Pp32 inhibited the expression of vinculin in HepG2 cells. In vivo primary experimental results showed that pp32 might inhibit HepG2 cells metastasis in nude mice. Compared to HepG2-pcDNA3.1 cells, the tumor size formed by HepG2-pp32 cells in the nude mice armpit had no significant differences.
Pc3M cell lines stable over-expressing pp32 (Pc3M-pp32) were obtained, pp32 could lead to shape changes of Pc3M cells, but did not affect their malignant phenotypes. Pp32 also led to promotion of the migration and adhesion of human prostate cancer cells Pc3M. The role of pp32 in cells migration of HepG2 and Pc3M cells is different, which was very interesting and gives us challenges.
Conclusions and analysis:
Pp32 is a multifunctional protein, which not only plays a role as a nuclear protein, but sometimes pp32 is located in the cytoplasm to take action. It is not only involved in tissue development, signal transduction, transcriptional regulation and other physiological processes, but also can promote cell apoptosis, inhibit tumor growth, inhibit the activity of PP2A and have other functions. Our studies showed that pp32 could inhibit cell migration characteristics of hepatoma carcinoma cells lines HepG2, but pp32 could stimulate Pc3M cell migration. Studies of pp32 regulating cell migration are not only conducive to the development of tumor biology, but also contribute to find new pathways to the treatment of human cancer.
引文
[1]、Malek SN, Katumuluwa AI, Pasternack GR, Identification and preliminary characterization of two related proliferation-associated nuclear nuclear phosphoproreins, J Biol Chem, 1990,265, 22: 13400-13409.
[2]、Matilla A, Radrizzani M, The Anp32 family of proteins containing leucine-rich repeats, Cer- ebellum, 2005, 4,1: 7-18.
[3]、Fink TM, Vaesen M, Kratzin HD, Lichter P, Zimmer M, Localization of the gene encoding the putative human HLA class II-associated protein (PHAPI) to chromosome 15q22.3-q23 by fluorescence in situ hybridization, Genomics, 1995, 29: 309-310.
[4]、Tseng-Hui Chen, Jonathan R.Brody, Feodor E., Roman Chen, Structure of pp32, an acidic nuclear protein which inhibits oncogene-induced formation of transformed foci, Molecular Biol- ology of the Cell, 1996 December, 7: 2045-2056.
[5]、Matsuoka, Taoka KM, Satozawa N, et al, A nuclear factor containing the leucine-rich repeats expresses in murine cerebellar neurons, Proc Natl Acad Sci USA, 1994, 91: 9670-74.
[6]、Trevor Huyton, Cynthia Wolberger, The crystal structure of the tumor suppressor protein pp32 (Anp32a): Structural insights into Anp32 family of proteins, Protein Science, 2007, 16: 1308-1315.
[7]、Opal Puneet, Garcia JJ, McCall AE, et al, Generation and Characterization of LANP/pp32 Null Mice, [J]. Mol Cell Biol, 2004, 24: 3140-49.
[8]、Matsubae M, Kurihara T, Tachibana T, et al, Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein, FEBS Lett, 2000, 468, 2-3: 171-175.
[9]、Brennan C.M., Gallouzi I.E., and Steitz J.A., Protein ligands to HuR modulate its interaction with target mRNAs in vivo, J. Cell Biol, 2000, 151:1-14.
[10]、Jiang X, Kim HE, Shu H, Zhao Y, Zhang H, Kofron J, Donnelly J, Burns D, Ng SC, Rose- nberg S, Wang X, Distinctive roles of PHAPI proteins and prothymosin-alpha in a deathregulatory pathway, Science, 2003, 299: 223-226.
[11]、Michelle M Hill, Colin Adrain, Patrick J Duriez, Analysis of the composition, assembly kinetics and activity of native Apaf-1 apoptosomes, The EMBO Journal, 2004, 23: 2134-2145.
[12]、Slee EA, Adrain C, Martin SJ, Executioner caspases-3, -6, and-7 perform distinct,non- redundant, roles during the demolition phase of apoptosis, J Biol Chem ,2001 ,276: 7320-7326
[13]、Slee EA, Harte MT, Kluck RM,Alnemri ES, Green DR, Martin SJ, Ordering the cytochro- me-c initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner, J Cell Biol, 1999, 144: 281-292.
[14]、Pan W, da Graca LS, Shao Y, Yin Q, Wu H, Jiang X, PHAPI/pp32 suppresses tumorigen- esis by stimulating apoptosis, J Biol Chem, 2009; 284: 6946-54.
[15]、Li M, Guo H, Damuni Z., Purification and characterization of two potent heat-stable inhib- itors of protein phosphatase 2A from bovine kidney, [J]. Biochemistry 1995, 34, 6: 1988-1996.
[16]、Li M, Makkinje A, Damuni Z., Molecular identification of I1PP2A, a novel potent heat- stable inhibitor protein of protein phosphatase 2A, J Biochemistry, 1996, 35, 22: 6998-7002.
[17]、Dingwall C, Laskey RA, Nuclear targeting sequence--a consensus?, Trends Biochem Sci, 1991, 16, 12: 478-481.
[18]、Ma WJ, Cheng S, Campbell C, et al, Cloning and characterization of HuR, a ubiquitously expressed Elav-like protein, J Biol Chem, 1996, 271,14: 8144-8151.
[19]、Brennan CM, Gallouzi IE, Steitz JA, Protein ligands to HuR modulate its interaction with target mRNAs in vivo, J Cell Biol, 2000, 15, 1: 1-13.
[20]、Seo SB, Macfarlan T, McNamara P, et al, The oncoprotein set/TAF-1-beta, an in hibitor of histone acetyltransferase, inhibits active demethylation of DNA, intergrating DNA methylation and transcriptional signaling, J Biol Chem, 2002, 277, 28: 14005-14010.
[21]、Schneider R, Bannister AJ, Weise C, et al, Direct binding of INHAT to H3 tails disrupted by modifications, J Biol Chem, 2004, 279, 23: 23859-23862.
[22]、Kutney SN, Hong R, Macfarlan T, et al, A signaling role of histone-binding proteins and INHAT subunits pp32 and Set/TAF-Ibeta in integrating chromatin hypoacetylation and transcri- ptional repression, J Biol Chem, 2004, 279, 29: 30850-30855.
[23]、Ulitzur N, Humbert M, Pfeffer SR., Mapmodulin: A possible modulator of the interaction of microtubule-associated proteins with microtubules, PNAS, 1997, 94, 10: 5084-5089.
[24]、Opal P, Garcia JJ, Propst F, et al, Mapmodulin/leucine-rich acidic nuclear proein binds the light chain of microtubule associated protein 1B and modulates neuritogenesis, J Biol Chem, 2003, 278, 36: 34691-34699.
[25]、Matilla A, Koshy BT, Cummings CJ, et al, The cerebellar leucine-rich acid nuclear protein interacts with ataxin-1,Nature, 1997, 389(6654): 974-978.
[26]、Brody JR, Kadkol SS, Hauer MC, et al, pp32 reduction induces differentiation of TSU-Pr1 cells,Am J Pathol, 2004, 16,1: 273-283.
[27]、Kadkol SS, Brody JR, Pevsner J, Bai J, Pasternack GR, Modulation of oncogenic potential by alternative gene use in humanprostate cancer, Nature Med, 1999, 5: 275-279.
[28]、Kadkol SS, EI Naga GA, Brody JR, et al, Expression of pp32 gene family members in bre- ast cancer, Breast Cancer Research and Treatment, 2001, 68: 65-73.
[29]、Zhang DJ, Cui CP, Wu CT, et al, Expression of pp32 in human hepatoma cell line HepG2 leads to morphological changes, Bull Acad Mil Med Sci, 2007, 31: 343-345.
[30]、廖子君,雷光焰,张冠军等,肿瘤转移学,陕西科学技术出版社, 2007, 1:1-500.
[31]、Department of Health, the Executive Yuan, R.O.C., Cancer Registry Annual Report in Taiwan area, 2007.
[32]、Huo TI, Lin HC, Huang YH, Wu JC, Chiang JH, Lee PC, et al, The model for end-stage li- ver disease-based Japan integrated scoring system may have a better predictive ability for patie- nts with hepatocellular carcinoma undergoing locoregional therapy, Cancer 2006, 107: 141-148.
[33] Hugo, H. et al, Epithelial-mesenchymal and mesenchymal-epithelial transition in carcinoma progression., J. Cell. Physiol, 2007, 213: 374-383.
[34]、Huber, M. A., Kraut, N. & Beug, H.,Molecular requirements for epithelial-mesenchymal transition during tumor progression, Curr Opin Cell Biol, 2005, 17: 548-558.
[1]、Malek SN, Katumuluwa AI, Pasternack GR, Identification and preliminary characterization of two related proliferation-associated nuclear nuclear phosphoproreins, J Biol Chem, 1990, 265, 22: 13400-13409.
[2]、Matsuoka, Taoka KM, Satozawa N, et al, A nuclear factor containing the leucine-rich repeats expresses in murine cerebellar neurons, Proc Natl Acad Sci USA, 1994, 9(21): 9670- 9674.
[3]、Opal Puneet, Garcia JJ, McCall AE, et al, Generation and Characterization of LANP/pp32 Null Mice, Mol Cell Biol, 2004, 24(8): 3140-3149.
[4]、Trevor Huyton, Cynthia Wolberger, The crystal structure of the tumor suppressor protein pp32(Anp32a): structural insights into Anp32 family of proteins, Protein Science, 2007, 16:1308-1315.
[5]、Matilla, A., Koshy, B.T., Cummings, C.J., Isobe, T., Orr, H.T., and Zoghbi, H.Y., The cerebellar leucine-rich acidic nuclear protein interacts with ataxin-1, Nature, 1997, 389: 974– 978.
[6]、Vaesen, M., Barnikol-Watanabe, S., Gotz, H., Awni, L.A., Cole, T.,Zimmermann, B., Kratzin, H.D., and Hilschmann, N., Purification and characterization of two putative HLA class II associated proteins: PHAPI and PHAPII, Biol. Chem. 1994, Hoppe Seyler 375: 113–126.
[7]、Ulitzur, N., Rancano, C., and Pfeffer, S.R., Biochemical characterization of mapmodulin, a protein that binds microtubule-associated proteins, J. Biol. Chem, 1997b., 272: 30577–30582.
[8]、Matsuoka, K., Taoka, M., Satozawa, N., Nakayama, H., Ichimura, T.,Takahashi, N., Yamakuni, T., Song, S.Y., and Isobe, T., A nuclear factor containing the leucine-rich repeats expressed in murine cerebellar neurons, Proc. Natl. Acad. Sci., 1994, 91: 9670–9674.
[9]、Li, M., Guo, H., and Damuni, Z., Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney, Biochemistry, 1995, 34: 1988–1996.
[10]、Li, M., Makkinje, A., and Damuni, Z., Molecular identification of I1PP2A, a novel potent heat-stable inhibitor protein of protein phosphatase2A, Biochemistry, 1996, 35: 6998–7002.
[11]、Fink TM, Vaesen M, Kratzin HD, Lichter P, Zimmer M, Localization of the gene encoding the putative human HLA class II-associated protein (PHAPI) to chromosome 15q22.3-q23 by fluorescence in situ hybridization, Genomics, 1995, 29: 309-310.
[12]、Tseng-Hui Chen, Jonathan R.Brody, Feodor E.Roman Chen, Structure of pp32,an acidic nuclear protein which inhibits oncogene-induced formation of transformed foci, Molecular Biology of the Cell, 1996 December, 7:2045-2056.
[13]、Puneet O, Garcia JJ, McCall AE, et al, Generation and charaterization LANP/pp32 null mice, [J].Mol Cell Biol, 2004, 24, 8: 3140-3149.
[14]、Trevor Huyton, Cynthia Wolberger, The crystal structure of the tumor suppressor protein pp32(Anp32a): structural insights into Anp32 family of proteins, Protein Science, 2007,16: 1308-1315.
[15]、Brennan, C.M., Gallouzi, I.E., and Steitz, J.A., Protein ligands to HuR modulate its interaction with target mRNAs in vivo, J. Cell Biol, 2000, 151: 1-14.
[16]、Kadkol SS, Brody JR, Pevsner J, Bai J, Pasternack GR, Modulation of oncogenic potential by alternative gene use in human prostate cancer, Nature Med, 1999, 5: 275-279.
[17]、Kadkol SS, EI Naga GA, Brody JR, et al, Expression of pp32 gene family members in breast cancer, Breast Cancer Research and Treatment, 2001, 68: 65-73.
[18]、Chen S, Li B, Grundke-Iqbal I, Iqbal K, I1PP2A affects tau phosphorylation via association with the catalytic subunit of protein phosphatase 2A, J Biol Chem, 2008, 283: 10513-21.
[19]、Pan W, da Graca LS, Shao Y, Yin Q, Wu H, Jiang X, PHAPI/pp32 suppresses tumorigenesis by stimulating apoptosis, J Biol Chem, 2009, 284: 6946-54.
[20]、Zachary T. Schafer, Amanda B. Parrish, Kevin M. Wright, Enhanced sensitivity to cytochrome c–induced apoptosis mediated by PHAPI in breast cancer cells, Cancer Research, 2006 February 15, 6:2210-2218.
[21]、Hanahan D, Weinberg RA., The hallmarks of cancer, Cell, 2000, 100: 57-70.
[22]、Jiang X, Kim HE, Shu H, Zhao Y, Zhang H, Kofron J, Donnelly J,Burns D, Ng SC, Rosenberg S, Wang X, Distinctive roles of PHAPI proteins and prothymosin-alpha in a death regulatory pathway. Science, 2003, 299: 223-226.
[23]、Michelle M Hill, Colin Adrain,Patrick J Duriez, Analysis of the composition, assembly kinetics and activity of native Apaf-1 apoptosomes, The EMBO Journal ,2004, 23, 2134-2145.
[24]、Slee EA, Adrain C, Martin SJ, Executioner caspases-3, -6, and-7 perform distinct, non-redundant, roles during the demolition phase of apoptosis, J Biol Chem, 2001, 276: 7320-7326.
[25]、Slee EA, Harte MT, Kluck RM, Alnemri ES, Green DR, Martin SJ,Ordering the cytochrome c-initiated caspase cascade:hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner, J Cell Biol ,1999, 144: 281-292.
[26]、Li M, Guo H, Damuni Z., Purification and characterization of two potent heat-stable inhib-itors of protein phosphatase 2A from bovine kidney, [J]. Biochemistry, 1995, 34, 6:1988-1996.
[27]、Li M, Makkinje A, Damuni Z., Molecular identification of I1PP2A, a novel potent heat- stable inhibitor protein of protein phosphatase 2A, [J].Biochamistry, 1996, 35, 22: 6998-7002.
[28]、Janssens V,Goris I., Protein phosphatase 2A:a highly regulated family of serine/threonine phosphatases im plicated in cell growth and signaling, Biochem J, 2001, 353: 417-439.
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[30]、Katarzyna Lechward1, Olubunmi S. Awotunde2, Wojciech OEwi1tek1 Protein phosphatase 2A: Variety of forms and diversity of functions, Acta Biochimica Polonica, 2001, 48, 4:921- 933.
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[33]、Fukukawa C, Tanuma N, Okada T, et al, pp32/I1PP2A negatively regulates the Raf-1/MEK/ERK pathway, Cancer Letters, 2005, 226: 155-160.
[34]、Yu LG, Packman LC, Weldon M, et al., Protein Phosphatase 2A, A negative regulator of the ERK signaling pathway, is activated by Tyrosine phosphorylation of putative HLA class II- associated protein I (PHAPI)/pp32 in response to the antiproliferative Lectin, Jacalin, J Biol Che- m, 2004, 279: 4137-41383.
[35]、Ulitzur N, Ranca?o C, Pfeffer SR. Biochemical Characterization of Mapmodulin, a Protein That Binds Microtubule-associated Proteins. The Journal of Biological Chemistry, 1997; 272: 30577-30582.
[36]、Puneet Opal, Jesus J. Garcia, Friedrich Propst, et al, Mapmodulin/Leucine-rich acidic nuclear protein binds the light chain of microtubule-associated protein 1B and modulates neuritogenesis, The Journal of Biological Chemistry, 2003; 278: 34691-34699.
[37]、Bai J, Brody JR, Kadkol SS, Pasternack GR: Tumor suppression and Potentiation by manipulation of pp32 expression, Oncogene, 2001, 20: 2153-2160.
[38]、Jonathan R. Brody, Shrihari S. Kadkol,M. Claire Hauer, et al, pp32 reduction inducesdifferentiation of TSU-Pr1 cells, American Journal of Pathology, January 2004, 164, 1: 273-283.
[39]、Seo SB, Macfarlan T, McNamara P,et al, The oncoprotein set/TAF-1-beta, an inhibitor of histone acetyltransferase, inhibits active demethylation of DNA, intergrating DNA methylation and transcriptional signaling, J Biol Chem,2002,277,28: 14005-14010.
[40]、Matsuoka, K., Taoka, M., Satozawa, N., Nakayama, H., Ichimura, T., TakahashiN., Yamak- uni, T., Song, S. Y., and Isobe T., A nuclear factor containing the leucine-rich repeats expresses in murine cerebellar neurons, Proc. Natl. Acad. Sci, 1994, 91: 9670-9674.
[41]、Timothy K. Williams., Christina L. Costantino, Nikolai A. Bildzukewicz, et al, pp32 (ANP32A) expression inhibits pancreatic cancer cell growth and induces gemcitabine resistance by disrupting HuR binding to mRNAs, Plos one,November 2010,5,11:15455-15466.
[2]、Matilla A, Radrizzani M, The Anp32 family of proteins containing leucine-rich repeats, Cer- ebellum, 2005, 4,1: 7-18.
[3]、Fink TM, Vaesen M, Kratzin HD, Lichter P, Zimmer M, Localization of the gene encoding the putative human HLA class II-associated protein (PHAPI) to chromosome 15q22.3-q23 by fluorescence in situ hybridization, Genomics, 1995, 29: 309-310.
[4]、Tseng-Hui Chen, Jonathan R.Brody, Feodor E., Roman Chen, Structure of pp32, an acidic nuclear protein which inhibits oncogene-induced formation of transformed foci, Molecular Biol- ology of the Cell, 1996 December, 7: 2045-2056.
[5]、Matsuoka, Taoka KM, Satozawa N, et al, A nuclear factor containing the leucine-rich repeats expresses in murine cerebellar neurons, Proc Natl Acad Sci USA, 1994, 91: 9670-74.
[6]、Trevor Huyton, Cynthia Wolberger, The crystal structure of the tumor suppressor protein pp32 (Anp32a): Structural insights into Anp32 family of proteins, Protein Science, 2007, 16: 1308-1315.
[7]、Opal Puneet, Garcia JJ, McCall AE, et al, Generation and Characterization of LANP/pp32 Null Mice, [J]. Mol Cell Biol, 2004, 24: 3140-49.
[8]、Matsubae M, Kurihara T, Tachibana T, et al, Characterization of the nuclear transport of a novel leucine-rich acidic nuclear protein-like protein, FEBS Lett, 2000, 468, 2-3: 171-175.
[9]、Brennan C.M., Gallouzi I.E., and Steitz J.A., Protein ligands to HuR modulate its interaction with target mRNAs in vivo, J. Cell Biol, 2000, 151:1-14.
[10]、Jiang X, Kim HE, Shu H, Zhao Y, Zhang H, Kofron J, Donnelly J, Burns D, Ng SC, Rose- nberg S, Wang X, Distinctive roles of PHAPI proteins and prothymosin-alpha in a deathregulatory pathway, Science, 2003, 299: 223-226.
[11]、Michelle M Hill, Colin Adrain, Patrick J Duriez, Analysis of the composition, assembly kinetics and activity of native Apaf-1 apoptosomes, The EMBO Journal, 2004, 23: 2134-2145.
[12]、Slee EA, Adrain C, Martin SJ, Executioner caspases-3, -6, and-7 perform distinct,non- redundant, roles during the demolition phase of apoptosis, J Biol Chem ,2001 ,276: 7320-7326
[13]、Slee EA, Harte MT, Kluck RM,Alnemri ES, Green DR, Martin SJ, Ordering the cytochro- me-c initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner, J Cell Biol, 1999, 144: 281-292.
[14]、Pan W, da Graca LS, Shao Y, Yin Q, Wu H, Jiang X, PHAPI/pp32 suppresses tumorigen- esis by stimulating apoptosis, J Biol Chem, 2009; 284: 6946-54.
[15]、Li M, Guo H, Damuni Z., Purification and characterization of two potent heat-stable inhib- itors of protein phosphatase 2A from bovine kidney, [J]. Biochemistry 1995, 34, 6: 1988-1996.
[16]、Li M, Makkinje A, Damuni Z., Molecular identification of I1PP2A, a novel potent heat- stable inhibitor protein of protein phosphatase 2A, J Biochemistry, 1996, 35, 22: 6998-7002.
[17]、Dingwall C, Laskey RA, Nuclear targeting sequence--a consensus?, Trends Biochem Sci, 1991, 16, 12: 478-481.
[18]、Ma WJ, Cheng S, Campbell C, et al, Cloning and characterization of HuR, a ubiquitously expressed Elav-like protein, J Biol Chem, 1996, 271,14: 8144-8151.
[19]、Brennan CM, Gallouzi IE, Steitz JA, Protein ligands to HuR modulate its interaction with target mRNAs in vivo, J Cell Biol, 2000, 15, 1: 1-13.
[20]、Seo SB, Macfarlan T, McNamara P, et al, The oncoprotein set/TAF-1-beta, an in hibitor of histone acetyltransferase, inhibits active demethylation of DNA, intergrating DNA methylation and transcriptional signaling, J Biol Chem, 2002, 277, 28: 14005-14010.
[21]、Schneider R, Bannister AJ, Weise C, et al, Direct binding of INHAT to H3 tails disrupted by modifications, J Biol Chem, 2004, 279, 23: 23859-23862.
[22]、Kutney SN, Hong R, Macfarlan T, et al, A signaling role of histone-binding proteins and INHAT subunits pp32 and Set/TAF-Ibeta in integrating chromatin hypoacetylation and transcri- ptional repression, J Biol Chem, 2004, 279, 29: 30850-30855.
[23]、Ulitzur N, Humbert M, Pfeffer SR., Mapmodulin: A possible modulator of the interaction of microtubule-associated proteins with microtubules, PNAS, 1997, 94, 10: 5084-5089.
[24]、Opal P, Garcia JJ, Propst F, et al, Mapmodulin/leucine-rich acidic nuclear proein binds the light chain of microtubule associated protein 1B and modulates neuritogenesis, J Biol Chem, 2003, 278, 36: 34691-34699.
[25]、Matilla A, Koshy BT, Cummings CJ, et al, The cerebellar leucine-rich acid nuclear protein interacts with ataxin-1,Nature, 1997, 389(6654): 974-978.
[26]、Brody JR, Kadkol SS, Hauer MC, et al, pp32 reduction induces differentiation of TSU-Pr1 cells,Am J Pathol, 2004, 16,1: 273-283.
[27]、Kadkol SS, Brody JR, Pevsner J, Bai J, Pasternack GR, Modulation of oncogenic potential by alternative gene use in humanprostate cancer, Nature Med, 1999, 5: 275-279.
[28]、Kadkol SS, EI Naga GA, Brody JR, et al, Expression of pp32 gene family members in bre- ast cancer, Breast Cancer Research and Treatment, 2001, 68: 65-73.
[29]、Zhang DJ, Cui CP, Wu CT, et al, Expression of pp32 in human hepatoma cell line HepG2 leads to morphological changes, Bull Acad Mil Med Sci, 2007, 31: 343-345.
[30]、廖子君,雷光焰,张冠军等,肿瘤转移学,陕西科学技术出版社, 2007, 1:1-500.
[31]、Department of Health, the Executive Yuan, R.O.C., Cancer Registry Annual Report in Taiwan area, 2007.
[32]、Huo TI, Lin HC, Huang YH, Wu JC, Chiang JH, Lee PC, et al, The model for end-stage li- ver disease-based Japan integrated scoring system may have a better predictive ability for patie- nts with hepatocellular carcinoma undergoing locoregional therapy, Cancer 2006, 107: 141-148.
[33] Hugo, H. et al, Epithelial-mesenchymal and mesenchymal-epithelial transition in carcinoma progression., J. Cell. Physiol, 2007, 213: 374-383.
[34]、Huber, M. A., Kraut, N. & Beug, H.,Molecular requirements for epithelial-mesenchymal transition during tumor progression, Curr Opin Cell Biol, 2005, 17: 548-558.
[1]、Malek SN, Katumuluwa AI, Pasternack GR, Identification and preliminary characterization of two related proliferation-associated nuclear nuclear phosphoproreins, J Biol Chem, 1990, 265, 22: 13400-13409.
[2]、Matsuoka, Taoka KM, Satozawa N, et al, A nuclear factor containing the leucine-rich repeats expresses in murine cerebellar neurons, Proc Natl Acad Sci USA, 1994, 9(21): 9670- 9674.
[3]、Opal Puneet, Garcia JJ, McCall AE, et al, Generation and Characterization of LANP/pp32 Null Mice, Mol Cell Biol, 2004, 24(8): 3140-3149.
[4]、Trevor Huyton, Cynthia Wolberger, The crystal structure of the tumor suppressor protein pp32(Anp32a): structural insights into Anp32 family of proteins, Protein Science, 2007, 16:1308-1315.
[5]、Matilla, A., Koshy, B.T., Cummings, C.J., Isobe, T., Orr, H.T., and Zoghbi, H.Y., The cerebellar leucine-rich acidic nuclear protein interacts with ataxin-1, Nature, 1997, 389: 974– 978.
[6]、Vaesen, M., Barnikol-Watanabe, S., Gotz, H., Awni, L.A., Cole, T.,Zimmermann, B., Kratzin, H.D., and Hilschmann, N., Purification and characterization of two putative HLA class II associated proteins: PHAPI and PHAPII, Biol. Chem. 1994, Hoppe Seyler 375: 113–126.
[7]、Ulitzur, N., Rancano, C., and Pfeffer, S.R., Biochemical characterization of mapmodulin, a protein that binds microtubule-associated proteins, J. Biol. Chem, 1997b., 272: 30577–30582.
[8]、Matsuoka, K., Taoka, M., Satozawa, N., Nakayama, H., Ichimura, T.,Takahashi, N., Yamakuni, T., Song, S.Y., and Isobe, T., A nuclear factor containing the leucine-rich repeats expressed in murine cerebellar neurons, Proc. Natl. Acad. Sci., 1994, 91: 9670–9674.
[9]、Li, M., Guo, H., and Damuni, Z., Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney, Biochemistry, 1995, 34: 1988–1996.
[10]、Li, M., Makkinje, A., and Damuni, Z., Molecular identification of I1PP2A, a novel potent heat-stable inhibitor protein of protein phosphatase2A, Biochemistry, 1996, 35: 6998–7002.
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