性激素受体与VEGF、EGFR及COX-2在结直肠癌肝转移中的作用

【作者】 罗吉辉；

【导师】 高枫； 张森；

【作者基本信息】 广西医科大学 ， 普通外科学， 2009， 博士

【摘要】 背景:结直肠癌是我国常见的消化道恶性肿瘤,近年来其发病率呈增高趋势,肝脏是结直肠癌最常见的转移部位,是结直肠癌患者死亡的主要原因。流行病学调查结果显示,各年龄段女性结直肠癌的患病率均低于男性,结直肠癌肝转移(CRC LM)患者男女性别比例为1.9:1,绝经后的妇女患结肠癌的风险较绝经前妇女高,应用雌激素替代疗法可以减少绝经期妇女患结肠癌的风险30%～40%。目前已经发现性激素参与调节肠粘膜细胞的分化和成熟,且雌激素受体(estrogen receptor, ER)和孕激素受体(progestogen receptor, PR)在大肠正常组织和肿瘤组织中均有表达,提示雌激素及孕激素可能影响这些肿瘤的发生与发展提供了客观依据。另外雌激素(单用或联用孕激素)替代疗法在临床上作为治疗结直肠癌的一种手段已取得一定疗效,由此推测CRC LM可能与性激素水平有关。目的:研究雌激素受体α、雌激素受体β、雄激素受体、孕激素受体在肠癌灶、正常肠粘膜及肝转移灶中的表达,探讨其在结直肠癌肝转移中的作用。材料和方法:病例入选标准:术前未经过放疗、化疗,术前一年之内无服用性激素药物史;剔除标准:切除的肝转移灶小,不能同时满足病理检查和实验研究需要量,收集2000年3月至2008年12月间CRC伴肝转移、并且除肝脏外无其他器官转移的病例,共43例,设为实验组,同时随机选取同期住院接受手术治疗的单纯性CRC病例43例,设为对照组,应用免疫组化SP法和SuperPicTureTM Polymer二步法测定———肠癌灶、正常肠粘膜及肝转移灶中雌激素受体(ERα、ERβ)、孕激素受体(PR)、雄激素受体(AR)的表达。另外收集2005年6月至2008年12月间住院手术治疗的结直肠癌肝转移病例和单纯性结直肠癌病例各30例,应用RT-PCR检测结直肠癌肿瘤组织和正常粘膜组织中ERβmRNA的表达情况。统计学方法:所得资料采用两样本率比较、Fisher确切概率法、χ2分割法计算多样本率的两两比较以及两小样本均数比较t检验处理,检验水准取α=0.05,由SPSS13.0 for windows软件包完成统计工作。结果:肠癌灶、正常肠粘膜及肝转移病灶均未检测到ERα蛋白表达。肝转移灶表达ERβ蛋白,其表达率13.2%(5/43),低于肠癌病灶ERβ蛋白表达率36.0%(31/86),差异有统计学意义(P<0.05),与肝转移组正常肠粘膜ERβ蛋白表达率60.5%(26/43)比较,差异有统计学意义(P<0.05)。实验组和对照组肠癌灶ERβ蛋白表达率分别为34.9% ,37.2% ,二者差异无统计学意义,P>0.05,两组正常肠粘膜ERβ蛋白表达率分别为60.5% ,62.8% ,二者差异无统计学意义,P>0.05。实验组和对照组肠癌灶ERβ蛋白表达率均明显低于正常肠粘膜,P<0.05。肝转移灶PR蛋白表达率4.7%(2/43),低于肠癌病灶PR蛋白表达率40.7%(35/86),差异有统计学意义(P<0.05),与肝转移组正常肠粘膜PR蛋白表达率55.8%(19/43)比较,差异有统计学意义(P<0.05)。实验组和对照组肠癌灶PR蛋白表达率分别为39.5%(17/43),41.9%(18/43),二者差异无统计学意义,P>0.05,两组正常肠粘膜PR蛋白表达率均为55.8%(14/43),无差异。实验组和对照组肠癌灶PR蛋白表达率均明显低于正常肠粘膜,P<0.05。肝转移灶中AR蛋白表达率7.0%低于肠癌病灶AR蛋白表达率27.9%(24/86),差异有统计学意义(P<0.05),与同组正常肠粘膜AR蛋白表达率23.3%比较,差异无统计学意义(P>0.05)。实验组和对照组肠癌灶AR蛋白表达率分别为23.3%(10/30),32.6%(14/30),二者差异无统计学意义,P>0.05,两组正常肠粘膜AR蛋白表达率分别为23.3%(10/30)和37.2%(16/30),差异无统计学意义。实验组和对照组肠癌灶和正常肠粘膜AR蛋白表达率差异无统计学意义。ERβmRNA在实验组肠癌组织中的表达率为33.3%(10/30),低于其该组正常肠粘膜中的表达率60%(18/30),差异有统计学意义,P<0.05,ERβmRNA在对照组肠癌组织中的表达率为43.3%(13/30),低于其该组正常肠粘膜中的表达率73.3%(22/30),差异有统计学意义,P<0.05,ERβmRNA在实验组和对照组肠癌灶或正常肠粘膜间表达差异无统计学意义,P>0.05。结论:ERβ蛋白是CRC肝转移灶中的优势ER,CRC肝转移灶和肠癌癌灶均未发现ERα蛋白表达。实验组中,肝转移灶、肠癌灶和正常肠粘膜的ERβ蛋白和PR蛋白表达率呈依次升高的趋势,ERβmRNA在结直肠癌与相应正常肠粘膜中的的表达也有类似趋势。随着肿瘤进展,ERβmRNA表达降低,由此所致ERβ蛋白逐渐减少,推测ERβ对于结直肠癌肝转移是保护因子。PR可能协同作用于ERβ并对结直肠癌肝转移起保护性作用。AR在结直肠癌中有低表达,其在结直肠癌肝转移中的作用尚不明确。背景:结直肠癌是常见消化道恶性肿瘤,发病率在男性恶性肿瘤中排第四位,女性排第三位,近年来,随着我国生活条件和饮食习惯的改变,结直肠癌发病率呈逐年上升趋势,且75%的病例发生于普通人群。虽然随着研究的深入和诊疗技术的提高,结直肠癌的早期诊断率和术后生存率有所提高,但仍未取得突破性进展,总体状况仍难令人满意。转移和复发是影响CRC预后的主要原因,肝脏是CRC最常见的转移部位,初诊时已有20%～40%的病人发生肝转移,此后20%～25%的患者发生异时性肝转移。提高CRC早期诊断率和治愈率是目前面对的一大难题,选择有效的预防药物及初步探讨其作用机制成为当前一项十分迫切的任务。目的:探讨VEGF和EGFR在结直肠癌肝转移中的作用及其与临床病理特征之间的关系。方法:按第一部分选择和剔除标准,随机选择43例结直肠癌肝转移患者作为实验组,43例结直肠癌无他处转移的患者作为对照组,取手术切除原发肠癌病灶和相应癌旁正常肠粘膜组织及肝转移组织,应用免疫组织化学检测VEGF和EGFR蛋白的表达情况,统计分析VEGF和EGFR在两组病例中肝转移病灶、肠癌病灶及正常肠粘膜中的表达差异,及其与肿瘤临床病理特征的关系。统计学方法:所得资料采用两样本率比较、Fisher确切概率法、χ2分割法计算多样本率的两两比较,不同组间阳性率差异比较用χ2检验,相关分析采用Spearman等级相关分析,检验标准P<0.05有统计学意义,用SPSS13.0 for windows软件包完成统计学分析。结果:(1)VEGF蛋白在实验组中肠癌病灶的阳性表达率为76.7%(33/43),高于该组的肝转移病灶的58.1%(25/43)和正常肠粘膜的9.3%(4/43),差异有统计学意义,P<0.05;实验组肠癌病灶VEGF蛋白的阳性表达率高于对照组的53.5%(23/43),差异有统计学意义,P<0.05;实验组和对照组正常肠粘膜VEGF蛋白率都比较低,分别为9.3%和0,差异无统计学意义,P>0.05;(2)EGFR蛋白在实验组中肠癌病灶的阳性表达率为79.1%(34/43),高于该组的肝转移病灶的48.8%(21/43)和正常肠粘膜的11.6%(5/43),差异有统计学意义,P<0.05;实验组肠癌病灶EGFR蛋白的阳性表达率高于对照组的58.1%(25/43),差异有统计学意义,P<0.05;实验组和对照组正常肠粘膜EGFR蛋白率都比较低,分别为11.6%(5/43)和0,差异无统计学意义,P>0.05;(3)将两组病例综合,分析VEGF和EGFR蛋白表达率与结直肠癌临床病理特征的关系,发现VEGF和EGFR蛋白表达与性别、年龄、肿瘤部位、组织类型、浸润深度无相关性,与肿瘤分化程度呈负相关,与肝转移和淋巴结转移呈正相关,且两者有相关性。结论:(1)VEGF和EGFR异常表达是结直肠癌发生肝转移中的一个重要环节。(2)VEGF和EGFR可作为结直肠癌有无转移的生物学指标,有助于预测CRC肝转移的发生和判断CRC肝转移的预后。目的:应用免疫组织化学技术检测COX-在结直肠癌发生肝转移病例中的表达情况,探讨COX-2对结直肠癌肝转移的影响。方法:按照论文第一部分的入选标准和剔除标准,收集43例结直肠癌肝转移病例作为实验组,同时随机选取单纯性结直肠癌病例作为对照组。免疫组化SuperPic Ture TMPolymer二步法检测——肝转移灶、肠癌灶和正常肠粘膜内COX-2蛋白的表达情况。统计学方法:所得资料采用两样本率比较、Fisher确切概率法、χ2分割法计算多样本率的两两比较,不同组间阳性率差异比较用χ2检验,相关分析采用Spearman等级相关分析,检验标准P<0.05有统计学意义,用SPSS13.0 for windows软件包完成统计学分析。结果:肝转移灶中有较高COX-2蛋白表达率,为44.2%(19/43),但低于肠癌灶的79.1%(34/43),差异有统计学意义(P<0.05),实验组肠癌灶的蛋白表达率79.1%(34/43)远高于正常肠粘膜的11.6%(5/43),差异有显著意义,P<0.05,实验组肠癌灶COX-2蛋白表达率与对照组肠癌灶COX-2蛋白表达率65.1%(28/43)比较,无明显差异,P>0.05。将两组病例综合,分析COX-2蛋白表达率与临床病理因素的关系,发现COX-2蛋白过表达与肿瘤分化程度呈负相关,与淋巴结转移和肝转移呈正相关,而与性别、年龄、肿瘤部位、组织类型和浸润深度无明确相关关系。结论:COX-2蛋白在结直肠癌组织中过表达,尤其以肝转移组结直肠癌组织中的COX-2蛋白表达为高,可能在结直肠癌的发生发展中起到重要作用。1. COX-2蛋白表达水平与肿瘤的分化程度和淋巴结转移呈正相关。2. COX-2蛋白表达水平与患者的性别、年龄、肿瘤部位及浸润深度无明确相关性。3.可以根据COX-2的表达情况判断肿瘤的恶性程度,它在结直肠癌肝转移中的表达为其靶向治疗提供了理论依据,调控COX-2蛋白表达以防止肿瘤的应用前景更加普遍。更多还原

【Abstract】 BACKGROUND: Colorectal cancer is a common gastrointestinal cancer and the incidence was increasing in China now. Liver is the most common metastatic site of colorectal cancer and the main reason of death from colorectal cancer. Epidemiological investigation showed that the incidence of colorectal cancer in women are lower than that in men(1.9:1). The risk of sufferring colon cancer in postmenopausal women is higher than in premenopausal women. Application of estrogen replacement therapy in postmenopausal women can reduce the risk of colon cancer by 30% ~ 40%. It has been found that sex steroid hormones regulated cell differentiation in intestinal mucosa and mature, and estrogen receptor and progesterone receptor express in normal colorectal tissue and tumor tissue. All of these indicate estrogen and progesterone affect these tumors’occurrence and development. In addition, estrogen (alone or combination with progesterone) replacement therapy in the clinical treatment of colorectal cancer have achieved a certain effect. It can be Speculated that CRC LM may associated with the level of sex hormones.OBJECTIVE: Study the expression of estrogen receptorα, estrogen receptorβ, androgen receptor, progesterone receptor in colorectal cancer tissues, normal tissues adjacent to cancer and liver metastasis tissues, discuss the roles play in colorectal cancer.Materials and Methods:Selection criteria: No pre-operative radiotherapy, chemotherapy. No taking hormone drugs within 1 year;Excluding criteria: lesions’size were too small to fit pathological examination and experimental studies. 43 cases of CRC with liver metastases and no metastasis in other organs from March 2000 to December 2008 were collorected as the experiment group, 43 cases of CRC with no metastases were colorected randomly at the same time as the control group, immunohistochemical SP method and the determination SuperPicTure TM Polymer two-step were used in the primary lesion of colorectal cancer, adjacent normal mucosa and estrogen receptor of liver metastasis (ERα, ERβ), progesterone receptor (PR ), androgen receptor (AR) expression. Statistical methods: The data were analyzed use a two-sample comparison, the fourfold table Fisher exact test,χ2 diverse segmentation method was used to calculate each two rates of varied rates, and t test comparing was used for two relatively small samples, the standard of admission testα= 0.05, by SPSS13.0 for statistical work has been completed windows package. The dates were analyzed using statistical software SPSS version13.0.RESULTS:No expression of ERαprotein were detected in colorectal lesions, normal mucosa and liver lesions. ERβprotein expression in liver lesions were 13.2% (5 / 43), less than in colorectal lesions 36.0% (31 / 86), the difference was statistically significant (P <0.05), and the rate of ERβprotein expression in normal mucosa was 60.5% (26/43, liver metastasis group), the difference was statistically significant (P <0.05). ERβprotein expression in colorectal cancer lesions between experimental and control group were 34.9%(15/43), 39.5%(17/43), the difference was not significant (P> 0.05), and in normal lesion were 60.5%(16/43), 62.8 %(27/43), the difference was not significant (P> 0.05). However, it was significant differencer between colorectal cancer and normal mucosa (P <0.05). PR protein expression in liver lesions were 4.7% (2 / 43), less than in colorectal lesions 40.7% (35/86), the difference was statistically significant (P <0.05), and the rate of PR protein expression in normal mucosa was 55.8% (24/43, liver metastasis group), the difference was statistically significant (P<0.05). PR protein expression in colorectal cancer lesions between experimental and control group were 39.5% (17/43), 41.9% (18/43), the difference was not significant (P> 0.05), and in normal lesion both group were 55.8% (24/43), there was no difference. However, it was significant differencer between colorectal cancer and normal mucosa (P <0.05). AR protein expression in liver lesions were 7.0% (3/43), less than in colorectal lesions 27.9% (24/86), the difference was statistically significant (P <0.05), and the rate of AR protein expression in normal mucosa was 23.3% (10/43, liver metastasis group), the difference was not significant (P> 0.05). AR protein expression in colorectal cancer lesions between experimental and control group were 23.3% (10/43), 32.6% (14/43), the difference was not significant (P> 0.05), and in normal lesion were 23.3% (10/43), 37.2 % (16/43), the difference was not significant (P> 0.05). H0were, there was no significant difference between colorectal cancer and nomarl mucosa. In experiment group, ERβmRNA expression in colorectal cancer lesions were 33.3% (10/30), less than in normal mucosa 60%(18/30), the difference was statistically significant (P <0.05). In control group, ERβmRNA expression in colorectal cancer lesions were 43.3%(13/30), less than in normal mucosa 73.3%(22/30), the difference was statistically significant (P <0.05). However, for ERβmRNA expression in colorectal cancer lesions or in normal mucosa, there were no difference between experiment group and control group (P>0.05).CONCLUSION:ERβprotein is the mainly ER in CRC liver metastasis. ERβprotein expressions were not found in CRC liver metastases and primary lesions. In experimental group, ERβprotein and PR protein expression rates was ascending in liver lesions, cancer lesions and normal mucosa orderly. Which indicated that ERβprotein is more invasive, ERβis a protective factor for colorectal liver metastases. PR synergized with ERβand plays a protective role in liver metastasis. AR had low expression in colorectal cancer. Background: Colorectal cancer is a common digestive tract cancer, the incidence of malignant tumors in men ranked fourth and women ranked the third in recent years, with the living conditions and dietary habits change, the incidence of colorectal cancer showed an upward trend year after year, and 75% of the cases occurred in the general population. Although the study with the depth and the improvement of medical technology, the early diagnosis of colorectal cancer and post-operative survival rate has increased, but has yet to achieve a breakthrough, the overall sentiment satisfactory situation. Metastasis and recurrence is the main reason for the impact of CRC prognosis, liver is the most common metastatic sites from CRC, more than 20%~40% of patients with synchronous liver metastases, and 20%~25% of patients with metachronous liver metastases . To improve the early diagnosis of CRC and the cure rate is currently a major problem, select an effective drug prevention and to explore its role in the mechanism has become a very urgent task. Objective: To investigate the VEGF and EGFR in liver metastases from colorectal cancer and its relationship with clinicopathological characteristics.Materials and Methods: According to the first part of the selection and exclusion criteria, 43 cases of randomly selected patients with liver metastases of colorectal cancer as the experimental group, 43 cases of colorectal cancer with no metastases as a control group, immunohistochemical detection of VEGF and EGFR protein expression were presented in primary cancer lesions and the corresponding adjacent normal mucosa and liver organization, statistical analysis of VEGF and EGFR in the two groups its relationship with tumor clinicopathological characteristics were performed. Statistical methods: The data were analyzed use a two-sample comparison, Fisher’s exact test,χ2 diverse segmentation method was used to calculate each two rates of varied rates,χ2 test was used for the comparison of the positive rate of different groups,correlation analysis Spearman correlated test , statistical significance was accepted at P<0.05, SPSS13.0 for statistical work has been completed windows package. The dates were analyzed using statistical software SPSS version13.0.Results: (1) VEGF protein in colorectal cancer lesions in the experimental group were 76.7 % (33/43), higher than the liver lesions and normal mucosa which were 58.1% (25/43) of 9.3% (4/43) ( P <0.05); VEGF protein expression in experimental group were higher than in control group at lesions of colorectal cancer (P <0.05); however, there were no significan difference at the normal mucosa between experimental group and control group (P> 0.05); (2) EGFR protein in colorectal cancer lesions in the experimental group were 79.1 %(34/43), higher than the liver lesions and normal mucosa which were 48.8% (21/43) of 11.6% (5/43) ( P <0.05); EGFR protein expression in experimental group were higher than in control group(58.1%, 25/43) at lesions of colorectal cancer(P <0.05); however, there were no significan difference at the normal mucosa between experimental group and control group(P> 0.05) ; (3) The relationship between VEGF and EGFR protein expression and clinicopathological characteristics found that VEGF and EGFR protein expression with gender, age, tumor site, histological type, depth of invasion associated with tumor differentiation was negatively correlated with liver metastasis and lymph node metastasis was positively correlated, and both are related to each other.Conclusions: (1) VEGF and abnormal expression of EGFR is the occurrence of liver metastasis of colorectal cancer in an important part. (2) VEGF and EGFR in colorectal cancer can be used as biological indicators, and can help to predict the occurrence of liver metastasis of CRC or CRC liver metastases to determine the prognosis for the future treatment of a new idea. Objective: to detect the affect of COX-2 in liver metastasis from colorectal cancer.Materials and Methods:43 cases with liver metastasis from colorectal cancer were selected as experimental group, and no-metastases cases of colorectal cancer were randomly selected as control group. Immunohistochemistry were used to discover the COX-2 protein expression in liver lesions, primary cancer and adjacent normal mucosa. Statistical methods: The data were analyzed use a two-sample comparison, Fisher’s exact test,χ2 diverse segmentation method was used to calculate each two rates of varied rates,χ2 test was used for the comparison of the positive rate of different groups,correlation analysis Spearman correlated test , statistical significance was accepted at P<0.05, SPSS13.0 for statistical work has been completed windows package. The dates were analyzed using statistical software SPSS version13.0.Results: The liver lesions in the experimental group was in a higher COX-2 protein expression rate (19/43,44.2%), but lower than colorectal cancer lesions (79.1%, 34/43,), the difference was statistically significant (P <0.05), and expression of COX-2 protein at colorectal cancer site were higher than at normal mucosa (11.6%, 5/43), the difference was significant( P <0.05). there were no significant difference between experimental group and control group at colorectal cancer site(65.1%, 28/43,control group ) (P> 0.05). Two cases will be a comprehensive analysis of COX-2 protein expression rate and the relationship between clinicopathological factors and found that COX-2 protein expression and tumor differentiation was negatively correlated with lymph node metastasis and liver metastasis was positively correlated with gender, age, tumor site, histological type and depth of invasion is no clear correlation.Conclusion: 1.COX-2 protein in colorectal cancer over-expressed, in particular colorectal cancer liver metastases group of COX-2 protein expression was higher in colorectal cancer may be the occurrence and development play an important role.2. COX-2 protein expression level and the degree of tumor differentiation and lymph node metastasis was positively correlated.3. COX-2 protein expression level and patient gender, age, tumor location and depth of invasion is no clear correlation.4. COX-2 expression can be the basis to determine liver metastasis from colorectal cancer and provides a theoretical basis for regulation and control COX-2 protein expression to prevent tumor metastases.更多还原