人骨肉瘤中PTEN、EGFR的表达及其与细胞增殖的关系
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摘要
骨肉瘤(Osteosarcoma)是最常见的骨恶性肿瘤，占骨原发恶性肿瘤的20％，其生物学行为多呈高度恶性，早期即易发生血道转移。虽然大剂量化疗的临床应用使骨肉瘤的5年生存率达80％，但仍有相当病人死于肺转移。生长因子通过与特异性受体结合，激活受体后信号途径，对调节细胞的增殖、存活有重要作用。但大量产生正性生长因子或其受体可通过自(旁)分泌机制使细胞增殖失控，在多种肿瘤的发生中起重要作用。多数生长因子在细胞内的第二信号均为3，4，5-三磷酸磷脂酰肌醇(phosphatidylinositol3，4，5-triphosphate，PIP3)，PIP3可与其它激酶结合促使细胞进入分裂周期。抑癌基因与细胞骨架蛋白tensin同源的在肿瘤的10号染色体有缺失的磷脂酶(phosphatase and tensin homolog deleted on chromosome ten,PTEN)是一种蛋白酪氨酸激酶，可使PIP3去磷酸化成PIP2，逆转PIP3的作用，抑制多数生长因子介导的细胞增殖。细胞增殖核抗原(Proliferation cell nuclear antigen，PCNA)是DNA聚合酶δ的辅助蛋白，可评价细胞的增殖状态。表皮生长因子受体(epidermal growth factor receptor，EGFR)和PTEN在骨肉瘤中的表达研究国内外少见报道。
目的：探讨PTEN、EGFR及PCNA在人骨肉瘤中表达的生物学意义及其互相关系，为阐明骨肉瘤的发病分子机制和基因治疗提供一定依据。
材料与方法：选取30例人骨肉瘤标本(所有标本均根据细胞和组织分化程度，将骨肉瘤分为三级：Ⅰ级7例，Ⅱ级14例，Ⅲ级9例)，以10例骨软骨瘤为对照，采用免疫组化S-P法，检测PTEN、EGFR及PCNA在人骨肉瘤中表达。以P＜0.05作为差异有显著性的检验标准。
结果：
1．PCNA蛋白表达：1．1 骨软骨瘤组PCNA染色阳性率为20.0％(2/10)，其中阴
2003年郑州大学硕士研究生毕业论文人骨肉瘤中PTEN、EGFR的表达及其与细胞增殖的关系
性(一)8例，弱阳性(+)2例;骨肉瘤组PCNA染色阳性率为100.0%(3。/30)，
其中弱阳性(+)13例，强阳性(++)17例。两组染色阳性率和染色强度均
有显著性差异(尸<0 .05)。1.2骨肉瘤I级组PCNA染色弱阳性(+)6例，
强阳性(++)1例;n、班级组PCNA染色弱阳性(+)7例，强阳性(++)
16例。两组差异有显著性(尸<0.05)。
2.PTEN蛋白表达:2.1骨软骨瘤组阳性表达率为90.0%(9/1的，其中阴性(一)
1例，弱阳性(+)2例，中度阳性(++)2例，强阳性(+十+)5例;骨肉
瘤组阳性表达率为70.0%(21/30)，其中阴性(一)9例，弱阳性(+)7例，中
度阳性(++)12例，强阳性(+++)2例。骨软骨瘤，骨肉瘤的染色阳性率
无显著性差异(P>0 .05)，染色强度有显著性差异(P<0 .05)。2.2骨肉瘤中
PTEN蛋白的表达2.2.1骨肉瘤I级组阳性表达率为85.7%(6/7)，其中阴性
(一)1例，弱阳性(+)3例，中度阳性(++)3例;n、m级组阳性表达率
为65.2%(15/23)，其中阴性(一)8例，弱阳性(+)4例，中度阳性(++)9
例，强阳性(+++)2例。n、nI级组与I级组染色阳性率和染色强度相比，
无显著性差异(尸>0 .05)。2.2.2骨肉瘤PCNA弱阳性组PTEN的阳性率为
76.9%(10/13)，其中阴性(一)3例，弱阳性(+)3例，中度阳性(++)6
例，强阳性(+++)1例;PCNA强阳性组PTEN的阳性率为64.7%(n/17)，
其中阴性(一)6例，弱阳性(+)4例，中度阳性(++)6例，强阳性(+++)
1例。PcNA弱阳性组与强阳性组PTEN染色阳性率与染色强度无显著性差
异(P>0 .05)。
3.EGFR蛋白表达:3.1骨软骨瘤组无阳性表达(0/10)，骨肉瘤组阳性表达率
为50.0%(15/30).骨软骨瘤、骨肉瘤染色阳性率有显著性差异(尸<0 .05)。3.2
骨肉瘤中EGFR蛋白的表达3.2.1骨肉瘤I级组阳性表达率为14.3%(1/7)，
11、m级组阳性表达率为60.9%(14/23).n、m级组与I级组染色阳性率相
比差异无显著性差异(P>0.05)。3.2.2骨肉瘤PCNA弱阳性组与强阳性组
EGFR阳性率分别为23.1%(3/13)和70.6%(12/17)，两组比较差异有显著性
(P<0 .05)。
结论:
1、PCNA在骨肉瘤与骨软骨瘤间，骨肉瘤各级间表达有显著性差异(P<0 .05)，提
2003年郑州大学硕士研究生毕业论文
人骨肉瘤中PTEN、EGFR的表达及其与细胞增殖的关系
示其可作为骨肉瘤定性、分级的指标;
2、PTEN染色强弱在骨软骨瘤与骨肉瘤间存在显著性差异(尸<0.05)，提示其在
骨肉瘤的发生中起到重要的作用;P花N在骨肉瘤各级间，PCNA弱阳性组、
强阳性组间表达无显著性差异(尸>0.05)，提示其不可作为骨肉瘤的分级、恶
性增殖指标;
3、EGFR在骨肉瘤与骨软骨瘤间，PCNA弱阳性组、强阳性组间表达有显著性
差异(尸<0.05)，提示其在骨肉瘤的发生及恶性增殖中起到一定的作用;EGFR
在骨肉瘤各级间表达无显著性差异(P>0.05)，提示其不可作为骨肉瘤的分级
指标。
Osteosarcoma (OS) is the most common primary malignant tumor of bone, comprising 20% of all such malignancies. Its biological behavior is always highly malignant. Its hematogenous metastasis occurs more frequently and early. Though clinic apply of high-dose chemotherapy makes the 5-year survival rate of OS climb to 80%, a significant percentage of patients die from lung metastasis. Growth factors (GFs) activate post-receptor way by combining special receptors, which is important to cells survival. The second messenger of most GFs is phosphatidylinositol 3, 4, 5-triphosphate (PIPS). PIPS can promote proliferation of cells by combining other kinases. Tumor suppressor phosphatase and tensin homolog deleted on chromosome ten (PTEN) is a kind of protein tyrosine phosphatase. It can dephosphate PIPS into PIP2, reverse the use of PIPS and refrain cells proliferation induced by most GFs. Proliferation cell nuclear antigen (PCNA) is affiliated protein of DNA ploymerase 6 . It can be used to evaluate the state of cell proliferation. So far, there are few reports on the expression of PTEN, epidermal growth factor receptor (EGFR) gene in human OS.
Objective: In order to evaluate the role of PCNA, PTEN and EGFR in OS as well as their interaction, their expression in OS is investigated. It is expected that the study will help us to elucidate the molecular mechanism of OS and provide theoretical foundation for genie therapy.
Materials and methods: Thirty specimens of human OS tissues (OST) were obtained. They were classified as three grades based on the grade of differentiation (I grade 7 cases, II grade 14 cases, III grade 9 cases). The control groups were 10 cases
osteochondroma tissues (OCT). Immunohistochemical S-P method was used to exam the expression of PCNA, PTEN and EGFR in OS. There was a significant difference when P<0.05. Results:
1. Expression of PCNA protein: 1.1 The positive immunostaining rate of PCNA in OCT was 20.0%(2/10), in which negative and strong stain were 8, 2, respectively; The positive immunostaining rate in OST was 100.0 %( 30/30), in which negative and strong stain were 13, 17, respectively. Comparing the staining rate and intensity of OCT and OST respectively, there were significant differences (P<0.05). 1.2 In grade I of OST, PCNA weak and strong positive staining were 6, 1, respectively. In grade II+III of OST, PCNA weak and strong positive staining were 7, 16, respectively. Comparing the staining intensity, there was significant difference between grade I and grade II+III of OST. (P<0.05).
2. Expression of PTEN protein: 2.1 In OCT, the positive immunostaining rate of PTEN was 90.0%(9/10), in which negative, weak, moderate and strong immunostaining were 1, 2, 2, 5, respectively. In OST, the positive immunostaining rate of PTEN was 70.0 %( 21/30), in which negative, weak, moderate and strong immunostaining were 9, 7, 12, 2, respectively. Comparing the staining rate, there was no significant difference (P>0.05). Comparing the staining intensity, there was significant difference (P<0.05). 2.2 Expression of PTEN protein in OST: 2.2.1 In grade I of OST, the positive immunostaining rate of PTEN was 85.7%(6/7), in which negative, weak and moderate immunostaining were 1, 3, 3, respectively. In grade II+III of OST, the positive immunostaining rate of PTEN was 65.2%(15/23), in which negative, weak, moderate and strong immunostaining were 9, 4, 9, 2, respectively. Comparing the staining rate and intensity, there were no significant differences (P>0.05). 2.2.2 In PCNA weak positive staining group, the positive immunostaining rate of PTEN was 76.9%(10/13), in which negative, weak, moderate and strong immunostaining were 3, 3, 6, 1, respectively. In PCNA strong positive staining group, the positive immunostaining rate of PTEN was 64.7 %(11/17), in which negative, weak, moderate and strong immunostaining
were 6, 4, 6, 1, respectively. Comparing the staining rate and intensity of PTEN, there were no significant differences (P>0.05).
3. Expression of EGFR prot
目的：探讨PTEN、EGFR及PCNA在人骨肉瘤中表达的生物学意义及其互相关系，为阐明骨肉瘤的发病分子机制和基因治疗提供一定依据。
材料与方法：选取30例人骨肉瘤标本(所有标本均根据细胞和组织分化程度，将骨肉瘤分为三级：Ⅰ级7例，Ⅱ级14例，Ⅲ级9例)，以10例骨软骨瘤为对照，采用免疫组化S-P法，检测PTEN、EGFR及PCNA在人骨肉瘤中表达。以P＜0.05作为差异有显著性的检验标准。
结果：
1．PCNA蛋白表达：1．1 骨软骨瘤组PCNA染色阳性率为20.0％(2/10)，其中阴
2003年郑州大学硕士研究生毕业论文人骨肉瘤中PTEN、EGFR的表达及其与细胞增殖的关系
性(一)8例，弱阳性(+)2例;骨肉瘤组PCNA染色阳性率为100.0%(3。/30)，
其中弱阳性(+)13例，强阳性(++)17例。两组染色阳性率和染色强度均
有显著性差异(尸<0 .05)。1.2骨肉瘤I级组PCNA染色弱阳性(+)6例，
强阳性(++)1例;n、班级组PCNA染色弱阳性(+)7例，强阳性(++)
16例。两组差异有显著性(尸<0.05)。
2.PTEN蛋白表达:2.1骨软骨瘤组阳性表达率为90.0%(9/1的，其中阴性(一)
1例，弱阳性(+)2例，中度阳性(++)2例，强阳性(+十+)5例;骨肉
瘤组阳性表达率为70.0%(21/30)，其中阴性(一)9例，弱阳性(+)7例，中
度阳性(++)12例，强阳性(+++)2例。骨软骨瘤，骨肉瘤的染色阳性率
无显著性差异(P>0 .05)，染色强度有显著性差异(P<0 .05)。2.2骨肉瘤中
PTEN蛋白的表达2.2.1骨肉瘤I级组阳性表达率为85.7%(6/7)，其中阴性
(一)1例，弱阳性(+)3例，中度阳性(++)3例;n、m级组阳性表达率
为65.2%(15/23)，其中阴性(一)8例，弱阳性(+)4例，中度阳性(++)9
例，强阳性(+++)2例。n、nI级组与I级组染色阳性率和染色强度相比，
无显著性差异(尸>0 .05)。2.2.2骨肉瘤PCNA弱阳性组PTEN的阳性率为
76.9%(10/13)，其中阴性(一)3例，弱阳性(+)3例，中度阳性(++)6
例，强阳性(+++)1例;PCNA强阳性组PTEN的阳性率为64.7%(n/17)，
其中阴性(一)6例，弱阳性(+)4例，中度阳性(++)6例，强阳性(+++)
1例。PcNA弱阳性组与强阳性组PTEN染色阳性率与染色强度无显著性差
异(P>0 .05)。
3.EGFR蛋白表达:3.1骨软骨瘤组无阳性表达(0/10)，骨肉瘤组阳性表达率
为50.0%(15/30).骨软骨瘤、骨肉瘤染色阳性率有显著性差异(尸<0 .05)。3.2
骨肉瘤中EGFR蛋白的表达3.2.1骨肉瘤I级组阳性表达率为14.3%(1/7)，
11、m级组阳性表达率为60.9%(14/23).n、m级组与I级组染色阳性率相
比差异无显著性差异(P>0.05)。3.2.2骨肉瘤PCNA弱阳性组与强阳性组
EGFR阳性率分别为23.1%(3/13)和70.6%(12/17)，两组比较差异有显著性
(P<0 .05)。
结论:
1、PCNA在骨肉瘤与骨软骨瘤间，骨肉瘤各级间表达有显著性差异(P<0 .05)，提
2003年郑州大学硕士研究生毕业论文
人骨肉瘤中PTEN、EGFR的表达及其与细胞增殖的关系
示其可作为骨肉瘤定性、分级的指标;
2、PTEN染色强弱在骨软骨瘤与骨肉瘤间存在显著性差异(尸<0.05)，提示其在
骨肉瘤的发生中起到重要的作用;P花N在骨肉瘤各级间，PCNA弱阳性组、
强阳性组间表达无显著性差异(尸>0.05)，提示其不可作为骨肉瘤的分级、恶
性增殖指标;
3、EGFR在骨肉瘤与骨软骨瘤间，PCNA弱阳性组、强阳性组间表达有显著性
差异(尸<0.05)，提示其在骨肉瘤的发生及恶性增殖中起到一定的作用;EGFR
在骨肉瘤各级间表达无显著性差异(P>0.05)，提示其不可作为骨肉瘤的分级
指标。
Osteosarcoma (OS) is the most common primary malignant tumor of bone, comprising 20% of all such malignancies. Its biological behavior is always highly malignant. Its hematogenous metastasis occurs more frequently and early. Though clinic apply of high-dose chemotherapy makes the 5-year survival rate of OS climb to 80%, a significant percentage of patients die from lung metastasis. Growth factors (GFs) activate post-receptor way by combining special receptors, which is important to cells survival. The second messenger of most GFs is phosphatidylinositol 3, 4, 5-triphosphate (PIPS). PIPS can promote proliferation of cells by combining other kinases. Tumor suppressor phosphatase and tensin homolog deleted on chromosome ten (PTEN) is a kind of protein tyrosine phosphatase. It can dephosphate PIPS into PIP2, reverse the use of PIPS and refrain cells proliferation induced by most GFs. Proliferation cell nuclear antigen (PCNA) is affiliated protein of DNA ploymerase 6 . It can be used to evaluate the state of cell proliferation. So far, there are few reports on the expression of PTEN, epidermal growth factor receptor (EGFR) gene in human OS.
Objective: In order to evaluate the role of PCNA, PTEN and EGFR in OS as well as their interaction, their expression in OS is investigated. It is expected that the study will help us to elucidate the molecular mechanism of OS and provide theoretical foundation for genie therapy.
Materials and methods: Thirty specimens of human OS tissues (OST) were obtained. They were classified as three grades based on the grade of differentiation (I grade 7 cases, II grade 14 cases, III grade 9 cases). The control groups were 10 cases
osteochondroma tissues (OCT). Immunohistochemical S-P method was used to exam the expression of PCNA, PTEN and EGFR in OS. There was a significant difference when P<0.05. Results:
1. Expression of PCNA protein: 1.1 The positive immunostaining rate of PCNA in OCT was 20.0%(2/10), in which negative and strong stain were 8, 2, respectively; The positive immunostaining rate in OST was 100.0 %( 30/30), in which negative and strong stain were 13, 17, respectively. Comparing the staining rate and intensity of OCT and OST respectively, there were significant differences (P<0.05). 1.2 In grade I of OST, PCNA weak and strong positive staining were 6, 1, respectively. In grade II+III of OST, PCNA weak and strong positive staining were 7, 16, respectively. Comparing the staining intensity, there was significant difference between grade I and grade II+III of OST. (P<0.05).
2. Expression of PTEN protein: 2.1 In OCT, the positive immunostaining rate of PTEN was 90.0%(9/10), in which negative, weak, moderate and strong immunostaining were 1, 2, 2, 5, respectively. In OST, the positive immunostaining rate of PTEN was 70.0 %( 21/30), in which negative, weak, moderate and strong immunostaining were 9, 7, 12, 2, respectively. Comparing the staining rate, there was no significant difference (P>0.05). Comparing the staining intensity, there was significant difference (P<0.05). 2.2 Expression of PTEN protein in OST: 2.2.1 In grade I of OST, the positive immunostaining rate of PTEN was 85.7%(6/7), in which negative, weak and moderate immunostaining were 1, 3, 3, respectively. In grade II+III of OST, the positive immunostaining rate of PTEN was 65.2%(15/23), in which negative, weak, moderate and strong immunostaining were 9, 4, 9, 2, respectively. Comparing the staining rate and intensity, there were no significant differences (P>0.05). 2.2.2 In PCNA weak positive staining group, the positive immunostaining rate of PTEN was 76.9%(10/13), in which negative, weak, moderate and strong immunostaining were 3, 3, 6, 1, respectively. In PCNA strong positive staining group, the positive immunostaining rate of PTEN was 64.7 %(11/17), in which negative, weak, moderate and strong immunostaining
were 6, 4, 6, 1, respectively. Comparing the staining rate and intensity of PTEN, there were no significant differences (P>0.05).
3. Expression of EGFR prot
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25． 韦平，傅天云，冯捷，等．卵巢的子宫内膜样癌临床情况及DNA倍体分析与P53基因蛋白，PCNA表达测定的研究．现代妇产科进展．1997，6(3)：201-3．
26. van Dierendonck JH, Wijsman JH, Keijzer R, et al. Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdUrd labeling and Ki-67 staining. Am J Pathol. 1991, 138(5): 1165-72.
27． 费素娟，黄水平，周力新，等．食管癌组织中COX-2，P53和PCNA的表达及意义．中国癌症杂志．2003，13(1)：17-20．
28． 仲伟霞，孙兰萍，蔡淑萍，等。结肠癌nm23-H_1、CD_(44)V_6、PCNA表达与肿瘤转移的关系．肿瘤防治研究．2002，29(5)：431-1．
29． 张春林，廖威明，李佛保，等．骨肉瘤中P21~(WAFI)蛋白，PCNA与Ki-67的表达及意义．中国癌症杂志．2001，11(2)：109-12．
30. Li DM, Sun H. TEP1, encoded by a candidate tumor suppressor locus, is a novel protein tyrosine phosphatase regulated by transforming growth factor beta. Cancer Res. 1997, 57(11): 2124-9.
31. Haas-Kogan D, Shalev N, Wong M, et al. Protein kinase B (PKB/Akt) activity is elevated in glioblastoma cells due to mutation of the tumor suppressor PTEN/MMAC. Curr Biol. 1998, 8(21): 1195-8.
32. Stambolic V, Suzuki A, de la Pompa JL, et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell. 1998, 95(1): 29-39.
33. Sun H, Lesche R, Li DM, et al. PTEN modulates cell cycle progression and cell survival by regulating phosphatidylinositol 3, 4, 5, -trisphosphate and Akt/protein kinase B signaling pathway. Proc Natl Acad Sci U S A.
1999, 96(11): 6199-204.
34. Chen ST, Yu SY, Tsai M, et al. Mutation analysis of the putative tumor suppression gene PTEN/MMAC1 in sporadic breast cancer. Breast Cancer Res Treat. 1999, 55(1): 85-9.
35. Garcia JM, Silva JM, Dominguez G, et al. Allelic loss of the PTEN region (10q23) in breast carcinomas of poor pathophenotype. Breast Cancer Res Treat. 1999, 57(3): 237-43.
36． 朱悦，王东煜，昌村春彦，等．肿瘤抑制基因PTEN在骨及软骨瘤中的突变．中华医学杂志．2001，81(12)：715-8．
37. Lin C, Meitner PA, Terek RM. PTEN mutation is rare in chondrosarcoma. Diagn Mol PathoL 2002, 11(1): 22-6.
38. Levine RA, Forest T, Smith C. Tumor suppressor PTEN is mutated in canine osteosarcoma cell lines and tumors. Vet Pathol. 2002, 39(3): 372-8.
39. Salvesen HB, MacDonald N, Ryan A, et al. PTEN methylation is associated with advanced stage and microsatellite instability in endometrial carcinoma. Int J Cancer. 2001, 91(1): 22-6.
40． 李航，张祥福，王川，等．肿瘤抑制基因PTEN在胃癌中的表达及其临床意义．实用肿瘤杂志．2002，17(5)：301-3．
41. Lu Z, Jiang (3, Blume-Jensen P, et al. Epidermal growth factor-induced tumor cell invasion and metastasis initiated by dephosphorylation and downregulation of focal adhesion kinase. Mol Cell Biol. 2001, 21(12): 4016-31.
42. Johannessen LE, Ringerike T, Molnes J, et al. Epidermal growth tactor receptor efficiently activates mitogen-activated protein kinase in HeLa cells and Hep2 cells conditionally defective in clathrin-dependent endocytosis. Exp Cell Res. 2000, 260(1): 136-45.
43. Denning MF, Dlugosz AA, Cheng C, et al. Cross-talk between epidermal growth factor receptor and protein kinase C during calcium-induced differentiation of keratinocytes. Exp Dermatol. 2000, 9(3): 192-9.
44. Gutacker C, Klock G, Diel P, et al. Nerve growth tactor and epidermal
growth factor stimulate clusterin gene expression in PC12 cells. Biochem J. 1999,339 (Pt 3): 759-66.
45. Xiao ZQ, Majumdar AP. Induction of transcriptional activity of AP-1 and NF-kappaB in the gastric mucosa during aging. Am J Physiol Gastrointest Liver Physiol. 2000, 278(6): G855-65.
46. Hansen LA, Woodson RL 2nd, Holbus S, et al. The epidermal growth factor receptor is required to maintain the proliferative population in the basal compartment of epidermal tumors. Cancer Res. 2000, 60(13): 3328-32.
47． 徐庆中，王大亮，广德宏，等．人脑星形细胞瘤EGF，TGFα及其受体EGFR的表达．诊断病理学杂志．2002，9(4)：207-9．
48. Fang MA, Kujubu DA, Hahn TJ. The effects of prostaglandin E2, parathyroid hormone, and epidermal growth factor on mitogenesis, signaling, and primary response genes in UMR 106-01 osteoblast-like cells. Endocrinology. 1992, 131(5): 2113-9.
49. Oda Y, Wehrmann B, Radig K, et al. Expression of growth factors and their receptors in human osteosarcomas. Immunohistochemical detection of epidermal growth tactor, platelet-derived growth factor and their receptors: its correlation with proliferating activities and p53 expression. Gen Diagn Pathol. 1995, 141(2): 97-103.
50． 屈洪涛，袁先厚，袁忠惠，等．PTEN基因及PCNA与胶质母细胞瘤的关系．实用癌症杂志．2001，16(3)：253-5．
51. Singh SP, Lipman J, Goldman H, et al. Loss or aleared subcellular localization of P27 in Barrett's associated adenocarcinoma. [J] Cancer Res. 1998, 58(15): 1730-5.
52. Harsh GR, Keating MT, Escobedo JA, et al. Platelet derived growth factor (PDGF) autocrine components in human tumor cell lines. J Neurooncol. 1990, 8(1): 1-12.
53． 王东，肖华亮，李增鹏，等．骨肉瘤碱性纤维生长因子，转化生长因子
β及其受体与血管生成的关系．癌症，1999，18(2)：142—5．
54. Burrow S, Andrulis IL, Pollak M, et al. Expression of insulin-like growth factor receptor, IGF-1, and IGF-2 in primary and metastatic osteosarcoma. J Surg Oncol. 1998, 69(1): 21-7.
55． 胡守兴，张李，谢李，等．星形细胞瘤中EGFR、PCNA的表达及意义．湖南医学．2000；17(5)：374-5．
56． 陈玲，张明才，EGFR、C-myc与PCNA在大肠癌中的表达及意义，中国基层医药．2002，9(2)：107-9．
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24. Hall PA, Levison DA, Woods AL, et al. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990, 162(4): 285-94.
25． 韦平，傅天云，冯捷，等．卵巢的子宫内膜样癌临床情况及DNA倍体分析与P53基因蛋白，PCNA表达测定的研究．现代妇产科进展．1997，6(3)：201-3．
26. van Dierendonck JH, Wijsman JH, Keijzer R, et al. Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdUrd labeling and Ki-67 staining. Am J Pathol. 1991, 138(5): 1165-72.
27． 费素娟，黄水平，周力新，等．食管癌组织中COX-2，P53和PCNA的表达及意义．中国癌症杂志．2003，13(1)：17-20．
28． 仲伟霞，孙兰萍，蔡淑萍，等。结肠癌nm23-H_1、CD_(44)V_6、PCNA表达与肿瘤转移的关系．肿瘤防治研究．2002，29(5)：431-1．
29． 张春林，廖威明，李佛保，等．骨肉瘤中P21~(WAFI)蛋白，PCNA与Ki-67的表达及意义．中国癌症杂志．2001，11(2)：109-12．
30. Li DM, Sun H. TEP1, encoded by a candidate tumor suppressor locus, is a novel protein tyrosine phosphatase regulated by transforming growth factor beta. Cancer Res. 1997, 57(11): 2124-9.
31. Haas-Kogan D, Shalev N, Wong M, et al. Protein kinase B (PKB/Akt) activity is elevated in glioblastoma cells due to mutation of the tumor suppressor PTEN/MMAC. Curr Biol. 1998, 8(21): 1195-8.
32. Stambolic V, Suzuki A, de la Pompa JL, et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell. 1998, 95(1): 29-39.
33. Sun H, Lesche R, Li DM, et al. PTEN modulates cell cycle progression and cell survival by regulating phosphatidylinositol 3, 4, 5, -trisphosphate and Akt/protein kinase B signaling pathway. Proc Natl Acad Sci U S A.
1999, 96(11): 6199-204.
34. Chen ST, Yu SY, Tsai M, et al. Mutation analysis of the putative tumor suppression gene PTEN/MMAC1 in sporadic breast cancer. Breast Cancer Res Treat. 1999, 55(1): 85-9.
35. Garcia JM, Silva JM, Dominguez G, et al. Allelic loss of the PTEN region (10q23) in breast carcinomas of poor pathophenotype. Breast Cancer Res Treat. 1999, 57(3): 237-43.
36． 朱悦，王东煜，昌村春彦，等．肿瘤抑制基因PTEN在骨及软骨瘤中的突变．中华医学杂志．2001，81(12)：715-8．
37. Lin C, Meitner PA, Terek RM. PTEN mutation is rare in chondrosarcoma. Diagn Mol PathoL 2002, 11(1): 22-6.
38. Levine RA, Forest T, Smith C. Tumor suppressor PTEN is mutated in canine osteosarcoma cell lines and tumors. Vet Pathol. 2002, 39(3): 372-8.
39. Salvesen HB, MacDonald N, Ryan A, et al. PTEN methylation is associated with advanced stage and microsatellite instability in endometrial carcinoma. Int J Cancer. 2001, 91(1): 22-6.
40． 李航，张祥福，王川，等．肿瘤抑制基因PTEN在胃癌中的表达及其临床意义．实用肿瘤杂志．2002，17(5)：301-3．
41. Lu Z, Jiang (3, Blume-Jensen P, et al. Epidermal growth factor-induced tumor cell invasion and metastasis initiated by dephosphorylation and downregulation of focal adhesion kinase. Mol Cell Biol. 2001, 21(12): 4016-31.
42. Johannessen LE, Ringerike T, Molnes J, et al. Epidermal growth tactor receptor efficiently activates mitogen-activated protein kinase in HeLa cells and Hep2 cells conditionally defective in clathrin-dependent endocytosis. Exp Cell Res. 2000, 260(1): 136-45.
43. Denning MF, Dlugosz AA, Cheng C, et al. Cross-talk between epidermal growth factor receptor and protein kinase C during calcium-induced differentiation of keratinocytes. Exp Dermatol. 2000, 9(3): 192-9.
44. Gutacker C, Klock G, Diel P, et al. Nerve growth tactor and epidermal
growth factor stimulate clusterin gene expression in PC12 cells. Biochem J. 1999,339 (Pt 3): 759-66.
45. Xiao ZQ, Majumdar AP. Induction of transcriptional activity of AP-1 and NF-kappaB in the gastric mucosa during aging. Am J Physiol Gastrointest Liver Physiol. 2000, 278(6): G855-65.
46. Hansen LA, Woodson RL 2nd, Holbus S, et al. The epidermal growth factor receptor is required to maintain the proliferative population in the basal compartment of epidermal tumors. Cancer Res. 2000, 60(13): 3328-32.
47． 徐庆中，王大亮，广德宏，等．人脑星形细胞瘤EGF，TGFα及其受体EGFR的表达．诊断病理学杂志．2002，9(4)：207-9．
48. Fang MA, Kujubu DA, Hahn TJ. The effects of prostaglandin E2, parathyroid hormone, and epidermal growth factor on mitogenesis, signaling, and primary response genes in UMR 106-01 osteoblast-like cells. Endocrinology. 1992, 131(5): 2113-9.
49. Oda Y, Wehrmann B, Radig K, et al. Expression of growth factors and their receptors in human osteosarcomas. Immunohistochemical detection of epidermal growth tactor, platelet-derived growth factor and their receptors: its correlation with proliferating activities and p53 expression. Gen Diagn Pathol. 1995, 141(2): 97-103.
50． 屈洪涛，袁先厚，袁忠惠，等．PTEN基因及PCNA与胶质母细胞瘤的关系．实用癌症杂志．2001，16(3)：253-5．
51. Singh SP, Lipman J, Goldman H, et al. Loss or aleared subcellular localization of P27 in Barrett's associated adenocarcinoma. [J] Cancer Res. 1998, 58(15): 1730-5.
52. Harsh GR, Keating MT, Escobedo JA, et al. Platelet derived growth factor (PDGF) autocrine components in human tumor cell lines. J Neurooncol. 1990, 8(1): 1-12.
53． 王东，肖华亮，李增鹏，等．骨肉瘤碱性纤维生长因子，转化生长因子
β及其受体与血管生成的关系．癌症，1999，18(2)：142—5．
54. Burrow S, Andrulis IL, Pollak M, et al. Expression of insulin-like growth factor receptor, IGF-1, and IGF-2 in primary and metastatic osteosarcoma. J Surg Oncol. 1998, 69(1): 21-7.
55． 胡守兴，张李，谢李，等．星形细胞瘤中EGFR、PCNA的表达及意义．湖南医学．2000；17(5)：374-5．
56． 陈玲，张明才，EGFR、C-myc与PCNA在大肠癌中的表达及意义，中国基层医药．2002，9(2)：107-9．