| 结直肠癌中癌干细胞的细胞周期及病理学意义 |
| Alternative Title | The Cell Cycle of Cancer Stem Cells and Its Pathological Significance in Colorectal Carcinoma
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| 王芙蓉 |
| Subtype | 硕士
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| Thesis Advisor | 刘斌
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| 2011-05-26
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| Degree Grantor | 兰州大学
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| Place of Conferral | 兰州
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| Degree Name | 硕士
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| Keyword | 结直肠癌，肿瘤干细胞，癌干细胞 ，CD44 ，PCNA， P27， hMSH2， SW480 ， 细胞周期 ，干细胞 ，免疫组织化学染色 ，免疫组织化学双重染色， 组织芯片。
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| Abstract | 目的
探讨原发性结直肠癌组织中CD44蛋白、PCNA（S期标志物）、P27蛋白表达的关系及其临床病理学意义，了解CD44+结直肠癌干细胞的增殖状况及在细胞周期中所处的状态，并观察CD44+/PCNA﹣、CD44+/P27high结直肠癌干细胞的数量、分布、形态特征，为研究结直肠癌干细胞标记及其理论提供更多的实验依据。检测人高分化结直肠癌细胞系SW480中CD44、P27、PCNA的表达,分析其细胞周期及形态特点。最后比较干细胞因子hMSH2与癌干细胞CD44表达的差异，初步探讨CD44+结直肠癌干细胞的起源。
方法
取61例结直肠癌、10例正常肠粘膜组织及18例伴不典型增生的结直肠腺瘤制成42点的共三块组织芯片，应用免疫组化S-P法检测结直肠组织芯片中CD44、P27、PCNA及MSH2蛋白的表达。应用免疫组织化学双重染色方法，标记CD44+/PCNA﹣、CD44+/P27high、CD44+/ hMSH2+细胞，并观察其数量、形态及分布方式。应用免疫组化S-P法检测结直肠癌细胞系SW480细胞中CD44、P27、PCNA蛋白的表达，分析其所处的细胞周期，并观察阳性细胞的形态特点。
结果
1．在结直肠癌组织中，CD44+细胞数约占癌细胞数的0.1～55%（30.9±19.29），CD44+癌细胞形态有二种，一种核较大，染色质边集，呈空泡状，核膜核仁清晰，另一种核较小，染色质致密，呈小圆形，χ2检验显示结直肠癌组织中CD44的表达明显高于其在正常肠粘膜组织和腺瘤中的表达。在结直肠癌中，CD44的表达与淋巴结转移、分化程度及Dukes分期无关（P>0.05）。
2．在结直肠癌组织中，PCNA +细胞数约占癌细胞数85～98%（93±4.21），χ2检验显示结直肠癌组织中PCNA的表达明显高于其在正常肠粘膜组织和腺瘤中的表达，在结直肠癌组织中，PCNA的表达与淋巴结转移，分化程度及Dukes分期无关（P>0.05）。
3．结直肠癌组织中，PCNA+的细胞数远远多于CD44+的细胞数，CD44+细胞中90%以上的瘤细胞同时为PCNA+，只有0.1-6%（3.38±2.08）细胞PCNA﹣，极少数CD44+/PCNA－癌细胞胞浆较少，核致密深染，CD44+/PCNA－位于腺管基底侧、癌细胞之间或单个散在，呈点状、灶状分布。
4．在结直肠癌组织中，P27+细胞数约占癌细胞数0.1～35%（20.9±12.21），χ2检验显示结直肠癌组织中P27的表达低于其在正常肠粘膜组织和腺瘤中的表达，P27的表达与浸润深度、分化程度及Dukes分期无明显关系（P >0.05），P27蛋白在癌组织中表达强弱不一，强阳性细胞（P27high）呈灶性或单个散在分布，约占癌细胞总数的0.1～15%（9.79±4.68），CD44+/P27high占CD44+细胞数的0.1～5%（3.12±1.18）。
5．结直肠癌组织中，CD44+/P27high占CD44+细胞数的0.1～5%（3.12±1.18），位于癌细胞之间，主要以点状、灶状分布在腺癌组织中。
6. 人高分化结直肠癌细胞系SW480细胞有两种形态，一种为大多边形、胞浆丰富、大的不规则核、染色质粗糙；另一种为小圆形，胞浆较少，核致密深染；SW480细胞CD44、P27、PCNA阳性表达率分别为65.80±5.26%、31.28±5.14%、55±6.57%。SW480中可见少量处于G0/G1期的CD44+/PCNA－、CD44+/P27high的肿瘤细胞，CD44+/PCNA－细胞占CD44阳性细胞的比率为11±5.36%，CD44+/P27high细胞占CD44+细... |
| Other Abstract | To investigate the relationship between the expressions of CD44, PCNA (S-phase-marker), P27 and their clinic-pathologic significance in primary colorectal carcinoma, and to explore the state of proliferation and cell cycle of CD44+ colorectal cancer stem cells, furtherly to examine the quantity, distribution, morphological characteristics of CD44+/PCNA﹣, CD44+/P27high colorectal cancer stem cells (Co-CSCs), and to provide more evidence to colorectal cancer stem cells; To investigate the expressions of CD44, PCNA and P27 in human well-differentiated colorectal carcinoma cell line (SW480), and to analyse the cell cycle and the morphologic features of SW480; To compare the difference between the expression of CD44 and hMSH2 and to explore the genesis of CD44+ colorectal cancer stem cells.
Methods
61 cases of human colorectal carcinoma, 10 cases of normal mucosa and 18 cases of adenoma with atypical hyperplasia were collected and made into three tissue microarrays each containing 42 dots. Expressions of CD44, PCNA, P27 proteins were detected by SP (streptavidin HRP) immunohistochemical staining to these tissue microarrays. The location, quantity and distribution of CD44+/PCNA﹣,CD44+/P27high and CD44+/ hMSH2+ cells were detected by double immunohistochemical staining. The expressions of CD44, PCNA and P27 proteins of human well-differentiated colorectal carcinoma cell line (SW480) were detected by SP immunohistochemical staining to analyze the cell cycle and its morphologic features of SW480.
Results
1. In colorectal carcinoma, the rate of CD44+ tumor cells was about 0.1～55%（30.9±19.29）. There were two kinds of morphology of CD44+ tumor cells: one was bigger vacuolus nucleus, chromatin margination, clear nuclear membrane and nucleolus. Another one was smaller nucleus, chromatin dense. The expressions of CD44 in colorectal carcinoma were significantly higher than in normal mucosa and adenoma with atypical hyperplasia. Significant relationship was not found among the expression of CD44 and lymphatica metastasis, pathological grade and Dukes stage (P >0.05).
2. In colorectal carcinoma, the rate of PCNA+ tumor cells was about 85～98% (93±4.21). The expressions of PCNA in colorectal carcinoma were significantly higher than in normal mucosa and adenoma with atypical hyperplasia. There was no significant relationship among the expression of CD44 and lymphatical metastasis, pathological grade and Dukes stage (P >0.05).
3. In colorectal carcinoma, the amou... |
| URL | 查看原文
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| Language | 中文
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| Document Type | 学位论文
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| Identifier | https://ir.lzu.edu.cn/handle/262010/215926
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| Collection | 基础医学院
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Recommended Citation
GB/T 7714 |
王芙蓉. 结直肠癌中癌干细胞的细胞周期及病理学意义[D]. 兰州. 兰州大学,2011.
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