Oct4与P27、Skp2在乳腺病变中的表达及其相关性的研究
摘要
研究背景和目的:随着对干细胞和肿瘤起源研究的深入,人们认识到,干细胞的异常分化和增殖会导致肿瘤的发生。肿瘤的起源与干细胞基因的异常表达有着千丝万缕的联系。乳腺肿瘤虽然组织类型复杂,但其组织发生却有其同源性,都来自于乳腺上皮干细胞。干细胞癌变时,由于其故有潜能及微环境影响,可向不同表型分化,甚至转分化。
目前,利用不同的分离乳腺干细胞的方法已经得到了一些常用的乳腺干细胞的标记物,但对于特异的乳腺干细胞标记物尚在不断的探索之中。胚胎干细胞基因Oct-4在维持干细胞的未分化状态和调节干细胞的分化过程起着关键性的作用,参与干细胞的自我更新。研究表明,Oct4在干细胞,包括胚胎干细胞和成体干细胞中维持较高水平的表达,而在已分化组织中表达大幅度下降或缺如,可能是有用的干细胞标记物。
细胞周期研究取得的一些突破性进展使人们对细胞周期与肿瘤的发生、发展之间的关系的认识也不断加深。肿瘤也是一类细胞周期疾病,细胞增殖的失控是肿瘤组织最显著的特点。P27是一个新近发现的抑癌基因,是调节细胞周期的周期素激酶抑制因子,直接参与细胞的增殖和分化。Skp2(细胞S期激酶相关蛋白2)属于F-box蛋白家族,它通过作用于泛素-蛋白酶体系统进而调节P27的降解,在细胞周期调控中发挥重要作用。已有大量文献报道, P27的低表达和Skp2的过表达在多种恶性肿瘤发生发展中具有重要的作用。
肿瘤发生发展的干细胞学说和细胞周期调控学说都是国内外研究的热点,本课题致力于细胞周期调控因子P27、Skp2的表达强弱与乳腺上皮干细胞癌变关系的研究,为更深层次揭示乳腺病变发生发展及演变过程提供新的思路。
方法:
1.采用免疫组化SP法分别检测10例正常乳腺组织、20例乳腺增生症、37例浸润性乳腺癌中干细胞标记物Oct4、腺腔上皮标记物CK8的表达情况,探讨Oct4在乳腺各组织中的表达及其与分化的关系。
2.采用免疫组化SP法分别检测细胞周期调控蛋白P27、Skp2的表达情况,并对它们在乳腺正常组织、良性增生组织、乳腺肿瘤分化程度上的差异及它们各自之间的相关性进行观察总结。
结果:
1. Oct4在乳腺正常组织、增生、浸润癌组织中的表达逐渐增多,在此三种乳腺组织中的表达存在差异,P<0.05。乳腺癌中Oct4阳性表达率为43.2%,低分化的乳腺癌Oct4阳性表达率为100%,显著高于高分化的乳腺癌(18.2%),差异具有统计学意义(P<0.05)。浸润型导管癌1级、2级和3级Oct4阳性表达率分别为18.2%、45.5%、100%,差异有统计学意义(P<0.05)。
2. CK8在正常乳腺组织和乳腺增生病中均有表达,阳性率分别为90.0%、95.0%,37例乳腺癌中有25例表达,阳性率为67.6%,正常和增生组织与乳腺癌之间表达有明显差异(P<0.05)。
3. P27在正常乳腺组织、乳腺增生性疾病和浸润性导管癌中均有表达,其阳性率分别为90.0%、85.0%、51.4%,各组织类型之间的表达有显著的差异(P<0.01)。
4. Skp2在正常乳腺组织、乳腺增生性疾病、浸润性导管癌中均有表达,其阳性率分别为10.0%、20.0%、45.9%,各组织类型之间存在差异性(P<0.05)。
5.经Spearman相关分析发现:
乳腺癌中Oct4和CK8的表达呈现负相关,相关系数r=-0.242,P=0.048;乳腺癌中Skp2和P27的表达呈显著负相关,相关系数r=-0.665,P=0.00;乳腺癌中Oct4和Skp2的表达呈显著正相关,相关系数r=0.791,P=0.00
结论:
1. Oct4在乳腺癌组织中的表达水平高于正常乳腺组织和乳腺增生病,进一步证实了乳腺癌组织中可能存在具有自我更新和分化潜能的干细胞样细胞的理论研究。Oct4阳性乳腺癌细胞可能是正常乳腺干细胞向乳腺癌分化过程中的乳腺癌干细胞或“中间型细胞”。
2. Oct4在乳腺癌组织中表达与分化程度关系密切,提示Oct4与乳腺癌恶性程度有关,可能是评估乳腺癌预后的重要生物学指标之一。
3. Oct4在正常乳腺组织、增生组织和癌组织中表达呈逐渐增多趋势,即分化程度越低,表达越多;CK8作为腺腔上皮的标记物在正常乳腺组织和乳腺增生病中表达率高,在乳腺癌组织中的表达减少。Oct4与CK8的表达阳性数相关性分析显示,两者呈一定程度的负相关,说明乳腺上皮干细胞在向恶性程度逐渐转化过程中,细胞可能向腺腔上皮方向分化减弱,分化不成熟,分化程度下降。
4. Skp2在乳腺正常上皮组织低表达,增生组织中表达稍高,在乳腺癌中表达最高并随着肿瘤分化程度的下降,Skp2的表达逐渐升高。而P27则呈相反的表达趋势,两者呈现一定程度的负相关。Skp2的过度表达伴P27的低表达提示肿瘤具有更高的侵袭性及预后不良。
5. Oct4与Skp2在各乳腺组织类型中的表达显示,两者呈显著正相关,说明在乳腺干细胞癌变过程中,细胞周期调控因子Skp2的过表达可能促进了Oct4的表达,其机制可能是Skp2通过对P27的降解,使得乳腺干细胞发生转分化或者分化受阻,异常增殖,最终形成乳腺癌干细胞或“中间型细胞”。
Background and Objective: Along with deep research on the stem cells and the tumor origin, people realized that stem cell's dysdifferentiation and the multiplication can cause the occurrence of the tumor.There are very complicated relations between the tumor origin and the unusual expression of stem cell genes. Although the complex tissue type of mammary tumor , they all stemm from mammary stem cells , during the canceration,under the influence of inherent potential and the micro environment,the stem cells can differentiate to different phenotypes, even transdifferentiation.
At present, people used different methods to separate mammary stem cells and already obtained some commonly used mammary gland stem cell's markers, But to define special marker of mammary stem cell , people still need unceasing exploration. Embryonic stem cell gene Oct4 is playing the crucial role in maintenanceing undifferentiation condition and adjusting differentiation process of stem cells,as well as participating the self-renewal of stem cells. The research indicated that Oct4 maintains the high level expression ,including in the embryonic stem cell and the adult stem cell but it’s expression level descend greatly even lack in differentiated tissuues.So some researchers consider that it is the potential useful stem cell marker .
On the other hand,with the breakthrough of the study on cell cycle in recent years, the study about the relations between the cell cycle and the occurrence, development of the malignant tumor is going deeply into as well.Out of control of the cell cycle is the most remarkable characteristic of tumor tissues,which contributes to the emergence and development of carcinomas also, and tumors can be regard as a kind of cell cycle disease accordingly.
P27 is a newly found anti-oncogene which inhibit CDK and then inhibit the transition of cell cycle from G1 phase to S phase. Is adjusts the cell cycle the cyclical element activating enzyme inhibiting factor,Skp2 gene(whose expression product is Skp2 protein) is also protooncogene, which often overexpress in many tumor tissues and is related with differential degree of these tumors. Skp2 not only correlates with degradation of cell cycle regulation factors intensively,but also contributes to cell cycle regulation and cell proliferation.Expression of Skp2 is required for the ubiquitination and subsequent degradation of P27 and other cell cycle factors that it can regulate cell cycle. High Skp2 expression are detected in many tumors.
Stem cell theory and cell cycle regulation theory are all domestic and foreign research hot spots.This topic devotes to investigate the relation beween the expression of the cell cycle regulatory factor P27、Skp2 and the canceration of mammary stem cells ,which bring a new idea for preventing breast tumor and improving the therapy of breast cancer.
Methods:
1. 10 cases of normal mammary gland tissues, 20 cases of mammary gland proliferated diseases,37 cases of mammary gland tumors were investigated by using S-P method of immunohistory and analyzed the expression of Oct4 and CK8 in each tissue. And their differences in the types of normal、proliferated tissues , differentiated extent and respective dependability were also studied.
2. 10 cases of normal mammary gland tissues,20 cases of mammary gland proliferated diseases, 30 cases of mammary gland tumors were investigated by using S-P method of immunohistory and analyzed the expression of P27 and Skp2 in these tissues. And their differences in the types of normal、proliferated tissues , differentiated extent and respective dependability were also studied.
Results:
1. Oct4 expressions were detected in normal mammary gland tissues, proliferated and tumorr tissues, datas showed significant difference between the expressions of normal tissues and infiltrating carcinoma tissues and significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues (P<0.05).
2. There is 90% of normal mammary gland tissues and 95% hyperplastic tissues expressing CK8, in the breast carcinoma tissues was observed in 12 of 37(67.6%) cases, significant difference was found in normal as well as proliferated tissues and tumor tissues(P<0.05).
3. The expression of P27 protein in normal mammary gland tissues, proliferated and tumor tissues were 90.0%、85.0% and 51.4%, it showed significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues(P<0.01).
4. The positve rate of Skp2 protein in normal mammary gland tissues, proliferated and tumor tissues were 10.0%、20.0% and 45.9%, it showed significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues(P<0.05).
5. Spearman’s rank correlation test showed that Oct4 exprssion in breast carcinoma had negative correlation with CK8 expression.Correlation coefficient: r=0.242,P =0.048.
Skp2 exprssion in breast carcinoma had negative correlation with P27 expression.Correlation coefficient: r=-0.665,P=0.00.
Skp2 exprssion in breast carcinoma had negative correlation with Oct4 expression.Correlation coefficient: r=0.791,P=0.00.
Conclusion:
1. The significant differences between expressions of Oct4 implied that there were“stem-like cells”exisiting in the tissue of breast tumor.The Oct4-expressing cells in the breast carcinoma maybe the cancer stem cells of the breast carcinoma or“intermediate cells”.
2. Oct4 is probably relevant with differentiated extent and it might be an important index for evaluating the prognosis of breast tumor.
3. Oct4 expression in normal mammary gland tissues,proliferated tissues and cancerous tissues presents an increasing tendency. The lower differentiated extent is,the more expression occurs.As the marker of glandular cavity epithelium, CK8 was expressed highly in normal mammary gland tissues and cyclomastopathy, expression decrease in mammary cancer tissues, in each group there is difference in expressions. The results from correlation analysis showed that there was strong relation between the expression of Oct4 and of CK8,all the outcornes pointed out that Oct4 gene had some effect to the stem cell differentiation.
4. Skp2 expression low in normal mammary tissue,higer in proliferated tissues and higest in breast carcinoma. Skp2 became gradually higer in expression when the degree of tumor differentiation became poorer,but the expression of P27 presents the oppisite tendency. A negative correlation was observed between Skp2 and P27,over expression of Skp2 along with low expression of P27 may indicate a tumor with higer invasive tendency and poor prognosis.
5. The expressions of Oct4 and Skp2 were detected in various mammary gland tissues, datas showed significan positive correlation between them,which impled that during the canceration of mammary stem cells, the overexprssion of Skp2 may lead to the accelarated expression of Oct4.The possible mechanism is that P27 were degraded by Skp2,which blocked the differentiation of mammary stem cells. Finally, these unusual multiplication cells became breast cancer stem cells or“intermediate cells”.
目前,利用不同的分离乳腺干细胞的方法已经得到了一些常用的乳腺干细胞的标记物,但对于特异的乳腺干细胞标记物尚在不断的探索之中。胚胎干细胞基因Oct-4在维持干细胞的未分化状态和调节干细胞的分化过程起着关键性的作用,参与干细胞的自我更新。研究表明,Oct4在干细胞,包括胚胎干细胞和成体干细胞中维持较高水平的表达,而在已分化组织中表达大幅度下降或缺如,可能是有用的干细胞标记物。
细胞周期研究取得的一些突破性进展使人们对细胞周期与肿瘤的发生、发展之间的关系的认识也不断加深。肿瘤也是一类细胞周期疾病,细胞增殖的失控是肿瘤组织最显著的特点。P27是一个新近发现的抑癌基因,是调节细胞周期的周期素激酶抑制因子,直接参与细胞的增殖和分化。Skp2(细胞S期激酶相关蛋白2)属于F-box蛋白家族,它通过作用于泛素-蛋白酶体系统进而调节P27的降解,在细胞周期调控中发挥重要作用。已有大量文献报道, P27的低表达和Skp2的过表达在多种恶性肿瘤发生发展中具有重要的作用。
肿瘤发生发展的干细胞学说和细胞周期调控学说都是国内外研究的热点,本课题致力于细胞周期调控因子P27、Skp2的表达强弱与乳腺上皮干细胞癌变关系的研究,为更深层次揭示乳腺病变发生发展及演变过程提供新的思路。
方法:
1.采用免疫组化SP法分别检测10例正常乳腺组织、20例乳腺增生症、37例浸润性乳腺癌中干细胞标记物Oct4、腺腔上皮标记物CK8的表达情况,探讨Oct4在乳腺各组织中的表达及其与分化的关系。
2.采用免疫组化SP法分别检测细胞周期调控蛋白P27、Skp2的表达情况,并对它们在乳腺正常组织、良性增生组织、乳腺肿瘤分化程度上的差异及它们各自之间的相关性进行观察总结。
结果:
1. Oct4在乳腺正常组织、增生、浸润癌组织中的表达逐渐增多,在此三种乳腺组织中的表达存在差异,P<0.05。乳腺癌中Oct4阳性表达率为43.2%,低分化的乳腺癌Oct4阳性表达率为100%,显著高于高分化的乳腺癌(18.2%),差异具有统计学意义(P<0.05)。浸润型导管癌1级、2级和3级Oct4阳性表达率分别为18.2%、45.5%、100%,差异有统计学意义(P<0.05)。
2. CK8在正常乳腺组织和乳腺增生病中均有表达,阳性率分别为90.0%、95.0%,37例乳腺癌中有25例表达,阳性率为67.6%,正常和增生组织与乳腺癌之间表达有明显差异(P<0.05)。
3. P27在正常乳腺组织、乳腺增生性疾病和浸润性导管癌中均有表达,其阳性率分别为90.0%、85.0%、51.4%,各组织类型之间的表达有显著的差异(P<0.01)。
4. Skp2在正常乳腺组织、乳腺增生性疾病、浸润性导管癌中均有表达,其阳性率分别为10.0%、20.0%、45.9%,各组织类型之间存在差异性(P<0.05)。
5.经Spearman相关分析发现:
乳腺癌中Oct4和CK8的表达呈现负相关,相关系数r=-0.242,P=0.048;乳腺癌中Skp2和P27的表达呈显著负相关,相关系数r=-0.665,P=0.00;乳腺癌中Oct4和Skp2的表达呈显著正相关,相关系数r=0.791,P=0.00
结论:
1. Oct4在乳腺癌组织中的表达水平高于正常乳腺组织和乳腺增生病,进一步证实了乳腺癌组织中可能存在具有自我更新和分化潜能的干细胞样细胞的理论研究。Oct4阳性乳腺癌细胞可能是正常乳腺干细胞向乳腺癌分化过程中的乳腺癌干细胞或“中间型细胞”。
2. Oct4在乳腺癌组织中表达与分化程度关系密切,提示Oct4与乳腺癌恶性程度有关,可能是评估乳腺癌预后的重要生物学指标之一。
3. Oct4在正常乳腺组织、增生组织和癌组织中表达呈逐渐增多趋势,即分化程度越低,表达越多;CK8作为腺腔上皮的标记物在正常乳腺组织和乳腺增生病中表达率高,在乳腺癌组织中的表达减少。Oct4与CK8的表达阳性数相关性分析显示,两者呈一定程度的负相关,说明乳腺上皮干细胞在向恶性程度逐渐转化过程中,细胞可能向腺腔上皮方向分化减弱,分化不成熟,分化程度下降。
4. Skp2在乳腺正常上皮组织低表达,增生组织中表达稍高,在乳腺癌中表达最高并随着肿瘤分化程度的下降,Skp2的表达逐渐升高。而P27则呈相反的表达趋势,两者呈现一定程度的负相关。Skp2的过度表达伴P27的低表达提示肿瘤具有更高的侵袭性及预后不良。
5. Oct4与Skp2在各乳腺组织类型中的表达显示,两者呈显著正相关,说明在乳腺干细胞癌变过程中,细胞周期调控因子Skp2的过表达可能促进了Oct4的表达,其机制可能是Skp2通过对P27的降解,使得乳腺干细胞发生转分化或者分化受阻,异常增殖,最终形成乳腺癌干细胞或“中间型细胞”。
Background and Objective: Along with deep research on the stem cells and the tumor origin, people realized that stem cell's dysdifferentiation and the multiplication can cause the occurrence of the tumor.There are very complicated relations between the tumor origin and the unusual expression of stem cell genes. Although the complex tissue type of mammary tumor , they all stemm from mammary stem cells , during the canceration,under the influence of inherent potential and the micro environment,the stem cells can differentiate to different phenotypes, even transdifferentiation.
At present, people used different methods to separate mammary stem cells and already obtained some commonly used mammary gland stem cell's markers, But to define special marker of mammary stem cell , people still need unceasing exploration. Embryonic stem cell gene Oct4 is playing the crucial role in maintenanceing undifferentiation condition and adjusting differentiation process of stem cells,as well as participating the self-renewal of stem cells. The research indicated that Oct4 maintains the high level expression ,including in the embryonic stem cell and the adult stem cell but it’s expression level descend greatly even lack in differentiated tissuues.So some researchers consider that it is the potential useful stem cell marker .
On the other hand,with the breakthrough of the study on cell cycle in recent years, the study about the relations between the cell cycle and the occurrence, development of the malignant tumor is going deeply into as well.Out of control of the cell cycle is the most remarkable characteristic of tumor tissues,which contributes to the emergence and development of carcinomas also, and tumors can be regard as a kind of cell cycle disease accordingly.
P27 is a newly found anti-oncogene which inhibit CDK and then inhibit the transition of cell cycle from G1 phase to S phase. Is adjusts the cell cycle the cyclical element activating enzyme inhibiting factor,Skp2 gene(whose expression product is Skp2 protein) is also protooncogene, which often overexpress in many tumor tissues and is related with differential degree of these tumors. Skp2 not only correlates with degradation of cell cycle regulation factors intensively,but also contributes to cell cycle regulation and cell proliferation.Expression of Skp2 is required for the ubiquitination and subsequent degradation of P27 and other cell cycle factors that it can regulate cell cycle. High Skp2 expression are detected in many tumors.
Stem cell theory and cell cycle regulation theory are all domestic and foreign research hot spots.This topic devotes to investigate the relation beween the expression of the cell cycle regulatory factor P27、Skp2 and the canceration of mammary stem cells ,which bring a new idea for preventing breast tumor and improving the therapy of breast cancer.
Methods:
1. 10 cases of normal mammary gland tissues, 20 cases of mammary gland proliferated diseases,37 cases of mammary gland tumors were investigated by using S-P method of immunohistory and analyzed the expression of Oct4 and CK8 in each tissue. And their differences in the types of normal、proliferated tissues , differentiated extent and respective dependability were also studied.
2. 10 cases of normal mammary gland tissues,20 cases of mammary gland proliferated diseases, 30 cases of mammary gland tumors were investigated by using S-P method of immunohistory and analyzed the expression of P27 and Skp2 in these tissues. And their differences in the types of normal、proliferated tissues , differentiated extent and respective dependability were also studied.
Results:
1. Oct4 expressions were detected in normal mammary gland tissues, proliferated and tumorr tissues, datas showed significant difference between the expressions of normal tissues and infiltrating carcinoma tissues and significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues (P<0.05).
2. There is 90% of normal mammary gland tissues and 95% hyperplastic tissues expressing CK8, in the breast carcinoma tissues was observed in 12 of 37(67.6%) cases, significant difference was found in normal as well as proliferated tissues and tumor tissues(P<0.05).
3. The expression of P27 protein in normal mammary gland tissues, proliferated and tumor tissues were 90.0%、85.0% and 51.4%, it showed significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues(P<0.01).
4. The positve rate of Skp2 protein in normal mammary gland tissues, proliferated and tumor tissues were 10.0%、20.0% and 45.9%, it showed significant deviation existing in expressions of normal as well as proliferated tissues and tumor tissues(P<0.05).
5. Spearman’s rank correlation test showed that Oct4 exprssion in breast carcinoma had negative correlation with CK8 expression.Correlation coefficient: r=0.242,P =0.048.
Skp2 exprssion in breast carcinoma had negative correlation with P27 expression.Correlation coefficient: r=-0.665,P=0.00.
Skp2 exprssion in breast carcinoma had negative correlation with Oct4 expression.Correlation coefficient: r=0.791,P=0.00.
Conclusion:
1. The significant differences between expressions of Oct4 implied that there were“stem-like cells”exisiting in the tissue of breast tumor.The Oct4-expressing cells in the breast carcinoma maybe the cancer stem cells of the breast carcinoma or“intermediate cells”.
2. Oct4 is probably relevant with differentiated extent and it might be an important index for evaluating the prognosis of breast tumor.
3. Oct4 expression in normal mammary gland tissues,proliferated tissues and cancerous tissues presents an increasing tendency. The lower differentiated extent is,the more expression occurs.As the marker of glandular cavity epithelium, CK8 was expressed highly in normal mammary gland tissues and cyclomastopathy, expression decrease in mammary cancer tissues, in each group there is difference in expressions. The results from correlation analysis showed that there was strong relation between the expression of Oct4 and of CK8,all the outcornes pointed out that Oct4 gene had some effect to the stem cell differentiation.
4. Skp2 expression low in normal mammary tissue,higer in proliferated tissues and higest in breast carcinoma. Skp2 became gradually higer in expression when the degree of tumor differentiation became poorer,but the expression of P27 presents the oppisite tendency. A negative correlation was observed between Skp2 and P27,over expression of Skp2 along with low expression of P27 may indicate a tumor with higer invasive tendency and poor prognosis.
5. The expressions of Oct4 and Skp2 were detected in various mammary gland tissues, datas showed significan positive correlation between them,which impled that during the canceration of mammary stem cells, the overexprssion of Skp2 may lead to the accelarated expression of Oct4.The possible mechanism is that P27 were degraded by Skp2,which blocked the differentiation of mammary stem cells. Finally, these unusual multiplication cells became breast cancer stem cells or“intermediate cells”.
引文
1. Scholer HR, Dressler GR, Balling R, et al. Oct4: a germline-specific transcription facor mapping to the mouse t-complex. EMBO J, 1990, 9(7): 2185- 2195.
2. Scholer HR., Ruppert S, Suzuki N, et al. New type of POU domain in germ line-specific protein Oct4. Nature,1990,344(6265):435-439.
3. Odorico JS, Kaufman DS, Thomson JA. Multilineage differentiation from human embryonic stem cell lines. Stem cells, 2001,19(3):193-204.
4. Hubner K, Fuhrmann G., Christenson LK,et al. Derivation of oocytes from mouse embryonic stem cells. Science, 2003,300(5623):1251-1256.
5. Young HE, Black AC Jr. Adult stem cells. Anat Rec A Discov Mol Cell Evol Biol,2004,276(1):75-102.
6. Young H E, Duplaa C, Yost MJ, et al. Clonogenic analysis reveals reserve stem cells in postnatal mammals.Ⅱ. Pluripotent epiblast-like stem cells. Anat Rec A Discov Mol Cell Evol Biol,2004,277(1):178-203.
7. Dyce PW, Zhu H, Craiq J, et al. Stem cells with multilineage potential derived from porcine skin. Biochem Biophys Res Commun, 2004,316(3):651-658.
8. Jones TD, Ulbrigh TM, Eble JN, et al. Oct4 staining in testicular tumors : a sensitive and specific marker for seminoma and embryonal careinoma. Am J Surg Pathol, 2004, 28(7):935-940.
9. Jones TD,Ulbright TM,Eble JN,et al. OCT4: A sensitive and specific biomarker for intratubular germ cell neoplasia of the testis. Clin Cancer Res, 2004,10(24):8544-8547.
10. Cheng L,Thomas A,Roth LM,et al. OCT4: a novel biomarker for dysgerminona of the ovary. Am J Surg Pathol, 2004:28(10):1341-1346.
11. Hattab EM, Tu PH, Wilson JD, et al. OCT4 Immunohistoehemistry is superior to placental alkaline phosphatase(PLAP)in the diagnosis of central nervous system germinoma. Am J Surg Pathol, 2005, 29(3):368-371.
12. Tai MH, Chang CC, Kiupel M, et al. Oct4 expression in adult human stem cells: evidence in support of the stem cell theory carcinogenesis. Carcinogenesis, 2005, 26(2):495-502.
13. Blain SW, Scher HI, Cordon-Cardo C, et al. p27 as a target for cancer therapeutics.Cancer Cell,2003,3(2):111-115.
14. Juan Rosai,乳腺.回允中主译.阿克曼外科病理学,第八版.沈阳:辽宁教育出版社,1999,1625-1626.
15许良中,杨文涛.免疫组织化学反应结果的判定标准.中国癌症杂志,1996,6(4):229-231.
16. Kim CF, Jackson EL, Woolfenden AE, et al. Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell,2005,121(6):823-835.
17. Okamoto K, Okazawa H, Okuda A, et al. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells ,Cell,1990, 60(3): 461-472.
18. Scholer HR, Ruppert S, Suzuki N, et al. New type of POU domain in germ line-specific protein Oct-4,Nature,1990,344(6265):435-439.
19. Rosner MH, Viqano MA, Ozato K, et al. A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature,1990,345 (6277): 686-692.
20. Tai MH, Chang CC, Kiupel M, et al. Oct4 expression in adult human stem cells:evidence in support of the stem cell theory of carcinogenesis. Carcinogenesis, 2005, 26(2):495-502.
21. Hansis C, Grifo JA, Krey LC. Oct-4 expression in inner cell mass and trophectoderm of human blastocysts. Mol Hum Reprod,2000,6(11):999-1004
22. Alonso L, Fuchs E. Stem cells of the skin epithelium. Proc Natl Acad Sci USA, 2003, 10(suppl 1) :11830-11835
23. Akashi K, Traver D, Kondo M, et al. Lymphoid development from hematopietic stem cells.Int J Hematol, 1999,69(4):217-226.
24.卲世宏,姚运红.干细胞、肿瘤干细胞与肿瘤的关系.现代肿瘤医学, 2005,13(3): 430-432.
25. Perez-Losada J, Balmain A. Stem-cell hierarchy in skin cancer. Nat Rev Cancer, 2003, 3(6): 434-443.
26. Vogel G. Stem cell research. Rat brains repond to embryonic stem cells. Science, 2002, 295(5553):254-255.
27. Reya T, Morrison SJ, Clarke MF, et al. Stem cells, cancer and cancer stem cells, Nature,2001,414(6859):105-111.
28.马芳,王志彪.肿瘤,一个发育生物学的问题.生命科学,2005, 17(5):433-437.
29. Hattori N, Nishino K, Ko YG, et al. Epigenetic control of mouse Oct4 gene expression in embryonic stem cells and trophoblast stem cells. J Biol Chem, 2004,279(17):17063-17069.
30. Kiyokawa H, Kineman RD, Manova-Todorova KO, et al. Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27(Kip1). Cell, 1996, 85(5):721-732.
31. Carrano AC, Eytan E, Hershko A, et al. Skp2 is required for ubiquitin-mediated degradation of the CDK inhibitor P27. Nat Cell Biol, 1999, 1(4):193-199.
32.王华,王婧花. P27kip1与肿瘤.中华病理学杂志, 1999, 28(2):140-142 .
33.管晓翔,陈龙邦.肿瘤细胞p27 kip蛋白错位分布和低表达的分子机制.中华肿瘤防治杂志, 2006,13(10):786-789.
34. Guan X, Chen L, Wang J, et al. Mutations of phosphorylation sites Ser10 and Thr187 of p27 kip1abolish cytoplasmic redistribution but do not abrogate G0/1 phase arrest in the HepG2 cell line. Biochem Biophys Res Commun, 2006, 347(3):601-607.
35.王道协,文建国,索振河. Oct4在干细胞河肿瘤细胞中的表达.山东医药,2007, 47(8):75-77.
36. Malek NP, Sundberg H, McGrew S, et al. A mouse knock in model exposes sequential proteolytic pathways that regulate p27kip1 in G1 and S phase. Nature, 2001, 413 (6853):323-327.
1. Reya T, Clevers H. Wnt sigaling in stem cells and cancer. Nature, 2005, 434 (7035): 843-850
2. Nusse R. Wnt signaling in disease and in development. Cell Res, 2005,15(1): 28-32.
3. Liu X, Rubin JS, Kimmel AR. Rapid,Wnt-induced change in GSK3beta associations that regulate beta-catenin stabilization are mediated by Galpha proteins. Curr Biol,2005, 15(22):1989-1997.
4. Daniel DL, Weis WI. Beta-catenin directly displaces Groucho/TLE repressors from Tcf/Lef in Wnt-mediated transcription activation. Nat Struct Mol Biol, 2005,12(4): 364-371.
5. Koemer FC. Epithelial proliferation of ductal type. Semin Diagn Pathol, 2004,21(10):10-17
6. Chepko G, Smith GH. Ultrastructure of Three division-competent structurally- distinct cell populations contribute to murine mammary epithelial renewal. Tissue Cell, 2003, 35(2): 83-93.
7. Shackleton M, Vaillant F, Simpson KJ. Generation of a fuctional mammary gland from a single stem cell. Gene Dev , 2003, 17(10) :1253-1270.
8. Liu BY, Mcdermott SP, Khwaja SS, et al. The transforming activity of Wnt effectors correlates with their ability to induce the accumulation of mammary progenitor cells. Proc Natl Acad Sci USA, 2004, 101(12): 4158-4163.
9. Li Y, Welm B, Podsypaina K, et al. Evidence that transgenes encoding components of the Wnt signaling pathway preferentially induce mammary cancers from progenitor cells. Proc Natl Acad Sci USA, 2003, 100(26): 15853-15858.
10. Reya T, Morrison SI, Clarke MF, et al. Stem cells cancer and cancer stem cells. Nature, 2001, 414(25): 105-111.
11. Tepera SB, Mccrea PD, Rosen JM. A beta-catenin survival signal is required for nomal lobular development in the mammary gland. Cell Sci, 2003, 116(Pt6): 1137-1149.
12. Brennan KR, Brown AM. Wnt proteins in mammary development and cancer. Mammary Gland Biol Neoplasis, 2004, 9(2): 119-131.
13. Howe LR, Brown AM. Wnt signaling and breast cancer. Cancer Biol Ther, 2004, 3(1): 36-41.
14. Klopocki E, Kristiansen G, Wild PJ, et al. Loss of SFRP1 is associatied with breast cancer progression and poor prognosis in early stage tumors. Int J Oncol, 2004, 25(3): 641-649.
15. Jain M, Arvanitis C, Chu K, et al. Sustained loss of a neoplastic phenotype by brief inactivation of MYC. Science, 2002, 297(5578): 102-104.
2. Scholer HR., Ruppert S, Suzuki N, et al. New type of POU domain in germ line-specific protein Oct4. Nature,1990,344(6265):435-439.
3. Odorico JS, Kaufman DS, Thomson JA. Multilineage differentiation from human embryonic stem cell lines. Stem cells, 2001,19(3):193-204.
4. Hubner K, Fuhrmann G., Christenson LK,et al. Derivation of oocytes from mouse embryonic stem cells. Science, 2003,300(5623):1251-1256.
5. Young HE, Black AC Jr. Adult stem cells. Anat Rec A Discov Mol Cell Evol Biol,2004,276(1):75-102.
6. Young H E, Duplaa C, Yost MJ, et al. Clonogenic analysis reveals reserve stem cells in postnatal mammals.Ⅱ. Pluripotent epiblast-like stem cells. Anat Rec A Discov Mol Cell Evol Biol,2004,277(1):178-203.
7. Dyce PW, Zhu H, Craiq J, et al. Stem cells with multilineage potential derived from porcine skin. Biochem Biophys Res Commun, 2004,316(3):651-658.
8. Jones TD, Ulbrigh TM, Eble JN, et al. Oct4 staining in testicular tumors : a sensitive and specific marker for seminoma and embryonal careinoma. Am J Surg Pathol, 2004, 28(7):935-940.
9. Jones TD,Ulbright TM,Eble JN,et al. OCT4: A sensitive and specific biomarker for intratubular germ cell neoplasia of the testis. Clin Cancer Res, 2004,10(24):8544-8547.
10. Cheng L,Thomas A,Roth LM,et al. OCT4: a novel biomarker for dysgerminona of the ovary. Am J Surg Pathol, 2004:28(10):1341-1346.
11. Hattab EM, Tu PH, Wilson JD, et al. OCT4 Immunohistoehemistry is superior to placental alkaline phosphatase(PLAP)in the diagnosis of central nervous system germinoma. Am J Surg Pathol, 2005, 29(3):368-371.
12. Tai MH, Chang CC, Kiupel M, et al. Oct4 expression in adult human stem cells: evidence in support of the stem cell theory carcinogenesis. Carcinogenesis, 2005, 26(2):495-502.
13. Blain SW, Scher HI, Cordon-Cardo C, et al. p27 as a target for cancer therapeutics.Cancer Cell,2003,3(2):111-115.
14. Juan Rosai,乳腺.回允中主译.阿克曼外科病理学,第八版.沈阳:辽宁教育出版社,1999,1625-1626.
15许良中,杨文涛.免疫组织化学反应结果的判定标准.中国癌症杂志,1996,6(4):229-231.
16. Kim CF, Jackson EL, Woolfenden AE, et al. Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell,2005,121(6):823-835.
17. Okamoto K, Okazawa H, Okuda A, et al. A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells ,Cell,1990, 60(3): 461-472.
18. Scholer HR, Ruppert S, Suzuki N, et al. New type of POU domain in germ line-specific protein Oct-4,Nature,1990,344(6265):435-439.
19. Rosner MH, Viqano MA, Ozato K, et al. A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature,1990,345 (6277): 686-692.
20. Tai MH, Chang CC, Kiupel M, et al. Oct4 expression in adult human stem cells:evidence in support of the stem cell theory of carcinogenesis. Carcinogenesis, 2005, 26(2):495-502.
21. Hansis C, Grifo JA, Krey LC. Oct-4 expression in inner cell mass and trophectoderm of human blastocysts. Mol Hum Reprod,2000,6(11):999-1004
22. Alonso L, Fuchs E. Stem cells of the skin epithelium. Proc Natl Acad Sci USA, 2003, 10(suppl 1) :11830-11835
23. Akashi K, Traver D, Kondo M, et al. Lymphoid development from hematopietic stem cells.Int J Hematol, 1999,69(4):217-226.
24.卲世宏,姚运红.干细胞、肿瘤干细胞与肿瘤的关系.现代肿瘤医学, 2005,13(3): 430-432.
25. Perez-Losada J, Balmain A. Stem-cell hierarchy in skin cancer. Nat Rev Cancer, 2003, 3(6): 434-443.
26. Vogel G. Stem cell research. Rat brains repond to embryonic stem cells. Science, 2002, 295(5553):254-255.
27. Reya T, Morrison SJ, Clarke MF, et al. Stem cells, cancer and cancer stem cells, Nature,2001,414(6859):105-111.
28.马芳,王志彪.肿瘤,一个发育生物学的问题.生命科学,2005, 17(5):433-437.
29. Hattori N, Nishino K, Ko YG, et al. Epigenetic control of mouse Oct4 gene expression in embryonic stem cells and trophoblast stem cells. J Biol Chem, 2004,279(17):17063-17069.
30. Kiyokawa H, Kineman RD, Manova-Todorova KO, et al. Enhanced growth of mice lacking the cyclin-dependent kinase inhibitor function of p27(Kip1). Cell, 1996, 85(5):721-732.
31. Carrano AC, Eytan E, Hershko A, et al. Skp2 is required for ubiquitin-mediated degradation of the CDK inhibitor P27. Nat Cell Biol, 1999, 1(4):193-199.
32.王华,王婧花. P27kip1与肿瘤.中华病理学杂志, 1999, 28(2):140-142 .
33.管晓翔,陈龙邦.肿瘤细胞p27 kip蛋白错位分布和低表达的分子机制.中华肿瘤防治杂志, 2006,13(10):786-789.
34. Guan X, Chen L, Wang J, et al. Mutations of phosphorylation sites Ser10 and Thr187 of p27 kip1abolish cytoplasmic redistribution but do not abrogate G0/1 phase arrest in the HepG2 cell line. Biochem Biophys Res Commun, 2006, 347(3):601-607.
35.王道协,文建国,索振河. Oct4在干细胞河肿瘤细胞中的表达.山东医药,2007, 47(8):75-77.
36. Malek NP, Sundberg H, McGrew S, et al. A mouse knock in model exposes sequential proteolytic pathways that regulate p27kip1 in G1 and S phase. Nature, 2001, 413 (6853):323-327.
1. Reya T, Clevers H. Wnt sigaling in stem cells and cancer. Nature, 2005, 434 (7035): 843-850
2. Nusse R. Wnt signaling in disease and in development. Cell Res, 2005,15(1): 28-32.
3. Liu X, Rubin JS, Kimmel AR. Rapid,Wnt-induced change in GSK3beta associations that regulate beta-catenin stabilization are mediated by Galpha proteins. Curr Biol,2005, 15(22):1989-1997.
4. Daniel DL, Weis WI. Beta-catenin directly displaces Groucho/TLE repressors from Tcf/Lef in Wnt-mediated transcription activation. Nat Struct Mol Biol, 2005,12(4): 364-371.
5. Koemer FC. Epithelial proliferation of ductal type. Semin Diagn Pathol, 2004,21(10):10-17
6. Chepko G, Smith GH. Ultrastructure of Three division-competent structurally- distinct cell populations contribute to murine mammary epithelial renewal. Tissue Cell, 2003, 35(2): 83-93.
7. Shackleton M, Vaillant F, Simpson KJ. Generation of a fuctional mammary gland from a single stem cell. Gene Dev , 2003, 17(10) :1253-1270.
8. Liu BY, Mcdermott SP, Khwaja SS, et al. The transforming activity of Wnt effectors correlates with their ability to induce the accumulation of mammary progenitor cells. Proc Natl Acad Sci USA, 2004, 101(12): 4158-4163.
9. Li Y, Welm B, Podsypaina K, et al. Evidence that transgenes encoding components of the Wnt signaling pathway preferentially induce mammary cancers from progenitor cells. Proc Natl Acad Sci USA, 2003, 100(26): 15853-15858.
10. Reya T, Morrison SI, Clarke MF, et al. Stem cells cancer and cancer stem cells. Nature, 2001, 414(25): 105-111.
11. Tepera SB, Mccrea PD, Rosen JM. A beta-catenin survival signal is required for nomal lobular development in the mammary gland. Cell Sci, 2003, 116(Pt6): 1137-1149.
12. Brennan KR, Brown AM. Wnt proteins in mammary development and cancer. Mammary Gland Biol Neoplasis, 2004, 9(2): 119-131.
13. Howe LR, Brown AM. Wnt signaling and breast cancer. Cancer Biol Ther, 2004, 3(1): 36-41.
14. Klopocki E, Kristiansen G, Wild PJ, et al. Loss of SFRP1 is associatied with breast cancer progression and poor prognosis in early stage tumors. Int J Oncol, 2004, 25(3): 641-649.
15. Jain M, Arvanitis C, Chu K, et al. Sustained loss of a neoplastic phenotype by brief inactivation of MYC. Science, 2002, 297(5578): 102-104.