大肠癌原发灶及淋巴结转移灶中TS和Topo-Ⅱ表达的对比分析

目的探讨胸苷酸合成酶（TS）和拓扑异构酶II（Topo-Ⅱ）在大肠癌原发灶及淋巴结转移灶的表达及临床意义。方法应用组织芯片及免疫组化技术,检测TS和Topo-II在135例大肠癌和60例淋巴结转移灶的表达情况。结果 TS在135例大肠癌和癌旁正常黏膜组织中的阳性表达率分别为54.81%和42.96%,差异无统计学意义（P〉0.05）,大肠癌TS表达与各临床病理参数无关（P〉0.05）,60例大肠癌淋巴结转移灶中TS阳性表达率为41.67%,与原发灶TS阳性表达率（53.33%）比较差异无统计学意义（P〉0.05）及无相关性（r=0.045,P=0.732）。Topo-II在135例大肠癌和癌旁正常黏膜组织中的阳性表达率分别为53.33%和21.48%,差异有统计学意义（P〈0.05）,Topo-Ⅱ阳性表达率与大肠癌组织学分级有关,中分化腺癌高于高分化腺癌（P〈0.05）,60例大肠癌原发灶中Topo-Ⅱ阳性表达率（56.67%）显著高于淋巴结转移灶（31.67%,P〈0.05）且呈正相关（r=0.261,P=0.044）。结论大肠癌原发灶与淋巴结转移灶TS和Topo-Ⅱ蛋白表达存在一定的差异,淋巴结转移灶耐药蛋白表达的检测对术后体内残存肿瘤细胞耐药性评估可能更为客观。     

Correlations between expression levels of thymidylate synthase,thymidine phosphorylase and dihydropyrimidine dehydrogenase,and efficacy of 5-fluorouracil-based chemotherapy for advanced colorectal cancer

The efficacy of 5-fluorouracil (5-FU)-based chemotherapy for colorectal cancer (CRC) widely varies among patients; therefore, it is difficult to accurately predict chemotherapeutic responses. Some recent studies have found that key enzymes in the various metabolic pathways activated by 5-FU present potential predictors of treatment outcome. Of these enzymes, thymidylate synthase (TS), thymidine phosphorylase (TP), and dihydropyrimidine dehydrogenase (DPD) are known to play important roles in the efficacy of therapeutic agents. Here, we measured expression levels of TS, TP, and DPD in formalin-fixed, paraffin-embedded, CRC specimens and paracancerous tissue with normal mucosa by immunohistochemical and fluorescence real-time quantitative polymerase chain reaction techniques. We found no significant differences in TS, TP, and DPD expression levels between CRC specimens and paracancerous tissues (P > 0.05), although overall survival and the chemotherapeutic effect were relatively poor in CRC patients with relatively high expression levels of TS, TP, and DPD, as compared to those with comparatively low expression levels (P < 0.05). Therefore, TS, TP, and DPD mRNA levels appear to be suitable indicators of the efficacy of 5-FU-based chemotherapy and prognosis of CRC.     

Prognostic predictors in breast cancer patients with postoperative 5-fluorouracil-based chemotherapy

Thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) are enzymes involved in the metabolism of 5-fluorouracil (FU). To investigate the relationship of these activities with clinicopathological factors and survival, we measured TS (88 patients) and DPD (122 patients) activities in resected specimens of breast cancer by enzyme assay. Significant difference was found only in TS activity between tumors > or = 20 mm in diameter and those < 20 mm (p = 0.015). There were no significant differences in TS or DPD activity among any other factors. When patients were grouped based on the cut-off levels of TS and DPD activities, 5-year recurrence-free survival rate was 68.8% in the low TS group and 39.7% in the high TS group (p = 0.0081), and 50.8% in the low DPD group and 66.5% in the high DPD group (p = 0.1627). The Cox proportional hazard model demonstrated that in patients in whom we measured TS activity, significant prognostic factors were nodal status and estrogen receptor (ER) status by univariate analysis, and ER status was also significant by multivariate analysis. In patients in whom DPD activity was measured, the significant prognostic factor was ER status by univariate analysis, and ER and Progesterone receptor (PgR) status by multivariate analysis. These results suggested that TS activity, nodal status and hormone receptors may be possible predictors of clinical outcome in breast cancer, but further investigation on prognostic predictors in 5-FU-based chemotherapy is required.     

Population study of expression of thymidylate synthase and dihydropyrimidine dehydrogenase in patients with solid tumors

Thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) has been suggested to be sensitivity-limiting factors of 5-fluorouracil therapy in cancer patients. We conducted a large-scale population study on the activity of TS and DPD in patients with various solid tumors. A total of 2590 clinically removed tumors, consisting of 1112 colon, 724 gastric, 520 breast, and 236 non-small cell lung cancers, were provided to measure TS and DPD activity. TS activity in the gastric, colon, and non-small cell lung cancers was significantly higher than in matched non-cancerous tissue (P<0.0002), but there was no difference in TS expression between tumor and non-cancerous tissue from breast cancer patients. Gastric, breast, and non-small cell lung cancers showed significantly higher DPD activity than their corresponding non-cancerous tissues, but colon cancers did not. There was no correlation between TS activity and DPD activity, and thus each enzyme was considered to be an independent sensitivity-limiting factor for 5-fluorouracil therapy. The median TS activity and median DPD activity in all specimens including gastric, colorectal, breast, and non-small cell lung cancers tested were 0.041 and 110.1 pmol/mg protein, respectively. We classified each of the type of carcinoma into 4 groups by using the median activity of TS and DPD as the cutoff values: a low TS/low DPD group, high TS/low DPD group, low TS/high DPD group, and high TS/high DPD group. About 50% of the gastric, 47% of the colon, 70% of the breast and 30% of the non-small cell lung cancers had high TS activity, and 60% of the gastric, 40% of the colon, 48% of the breast, and 87% of the lung cancers had high DPD activity. Moreover, breast cancer was characterized by high TS activity and lung cancer by high DPD activity as compared with gastric and colon cancers, and their high activity levels may influence to the effectiveness of 5-fluorouracil against cancers of these organs. The results for expression of TS and DPD in clinically dissected tumors would be useful to estimate the efficacy of 5-fluorouracil in the treatment of cancer patients.     

Importance of hepatic artery node involvement in patients with colorectal liver metastases.

Hepatic artery lymph node (HALN) involvement is an adverse prognostic factor in patients treated for colorectal liver metastases. The prevalence of HALN positivity for mid-gut and hind-gut derived colonic tumours, for differing amounts of liver involvement, and for Dukes' A and B versus Dukes' C primary tumours was compared in 75 patients with colorectal liver metastases. All patients whose primary tumours did not invade lymph nodes (Dukes' A or B) had liver metastases that did not involve local hepatic nodes, regardless of the extent of the disease within the liver. This suggests that factors controlling metastasis are not identical with those which control lymphatic invasion in colorectal cancer. HALN positive patients may benefit less from treatment because they are significantly more likely to have both a greater burden of disease within the liver and a tumour with greater lymph invasive potential than patients with HALN negative liver metastases.     

Evaluation of Clinical Value of Single Nucleotide Polymorphisms of Dihydropyrimidine Dehydrogenase Gene to Predict 5-Fluorouracil Toxicity in 60 Colorectal Cancer Patients in China

Dihydropyrimidine dehydrogenase (DPD) activity could be affected by single nucleotide polymorphisms (SNPs), resulting in either no effect, partial or complete loss of DPD activity. To evaluate if SNPs of DPD can be used to predict 5-FU toxicity, we evaluated five SNPs of DPD (14G1A, G1156T, G2194A, T85C and T464A) by TaqMan real time PCR in 60 colorectal cancer patients. Clinical data demonstrated that there was higher correlation between DPD activity and toxic effects of 5-FU (p<0.05). Six patients were positive for G2194A detection, which were all heterozygous. Two patients had lower DPD activities (< 3) with higher toxic effects (≥stage III) while one patient was also positive for T85C detection. Ten patients were positive for T85C detection. Two patients were homozygous with lower DPD activities and higher toxic effects. Two patients were positive for the T464A detection, which were heterozygous with lower DPD activity and higher toxic effects and also positive for T85C detection. These data clearly indicated that the T464A and homozygous of the T85C are stronger biomarkers to predict the 5-FU toxicity. Our study significantly indicated that the detection for G2194A, T85C and T464A could predict ~13% of 5-FU severe toxic side effects.     

Comparative genomic hybridization analysis of primary colorectal carcinomas and their synchronous metastases

We have analyzed 26 tumors from 12 patients with metastatic colorectal adenocarcinoma by comparative genomic hybridization (CGH). Primary tumors and their lymph node metastases from five Dukes' C patients as well as primary tumors and their liver metastases from seven Dukes' D patients were used to assess the extent of genetic differences between primary and secondary colorectal carcinomas from the same patients, to calculate the degree of clonal divergence and genetic heterogeneity in metastatic colorectal cancer, and to determine the differences in genetic imbalances between Dukes' C and D stage tumors. We show that the same genetic aberrations were frequently found in the primary tumors and their metastases. However, metastases often contained genetic aberrations not found in the corresponding primary tumors. The comparison of Dukes' stages C and D revealed genetic aberrations common to both. However, reduced copy number of chromosome arm 17p (5/5 vs. 0/7; P = 0.001) was significantly associated with Dukes' stage C and lymph node metastases, while increased copy number of chromosome arms 6p (6/7 vs. 0/5; P = 0.007) and 17q (5/7 vs. 0/5; P = 0.027) was associated more with Dukes' stage D and liver metastases. Our results established a repertoire of chromosomal alterations associated with metastatic colorectal cancer and suggest that Dukes' C (lymph node metastasis) tumors are not always simply an earlier stage of Dukes' D (liver metastasis) tumors and, thus, in some instances at least, they are distinct forms of the disease.     

Dihydropyrimidine deshydrogenase (DPD): rhythm and consequences

Dihydropyrimidine deshydrogenase (DPD) is the rate limiting enzyme of 5-fluorouracil (5-FU) catabolism and its activity is generally determined in peripheral blood mononuclear cells. Several studies have highlighted interactions between toxicities to 5-FU and a DPD activity deficiency. Circadian variations in 5-FU anabolism enzymes are suggested. Circadian variations in 5-FU catabolism enzymes, and especially for DPD in healthy subjects or patients, have shown in some cases circadian variations in DPD activity but with different peak times. Based on this knowledge, chronomodulated therapy for the association 5-FU-folinic acid with maximal delivery rate in the first half of the night was shown clearly to be 5 times less toxic than control flat therapy. Nevertheless, in the most active chronotherapy pattern, 30% of the patients have also toxicities. However the timing of the individual peak of DPD activity remains controversial.     

Immunohistochemical expression of thymidylate synthase as predictor of response to capecitabine in patients with advanced colorectal adenocarcinoma

Capecitabine is an oral prodrug to 5-fluorouracil (5-FU). The primary target of 5-FU is thymidylate synthase (TS). A mainstay of colorectal adenocarcinoma chemotherapy is inhibition of TS, which may be one of many determinant factors when predicting the outcome of chemotherapies based on fluoropyrimidine treatment. This retrospective study included 39 patients with advanced colorectal adenocarcinoma treated with capecitabine. Response was assessed by measuring the amount of tumour in the course of treatment. TS expression was evaluated by scoring the immunohistochemical (IHC) reaction and assessing the predominant IHC reaction pattern. This study showed significant correlation between the predominant IHC reaction pattern and response, but no correlation between IHC score and response. The predominant IHC reaction pattern may be a useful parameter in prediction of clinical outcome in patients treated with capecitabine.     

人结直肠癌趋化因子配体生物轴及干细胞标志物基因表达与临床病理

背景：趋化因子配体12/趋化因子受体4生物学轴在肿瘤的特异性转移中有重要作用,而干细胞标志物糖蛋白激素受体5基因的表达对于肿瘤的增殖和侵袭转移发挥重要作用。 目的：观察趋化因子配体12/趋化因子受体4生物轴以及干细胞标志物糖蛋白激素受体5基因在人结直肠癌组织中的表达变化及其与临床中的病理特征的关系。 方法：收集2013年1至6月辽宁省肿瘤医院收治100名结直肠癌患者为实验组,100名健康体检者为对照组,采用免疫组织化学SP法检测两组组织中趋化因子配体12、趋化因子受体4及干细胞标志物糖蛋白激素受体5 mRNA表达情况,并分析趋化因子配体12、趋化因子受体4及干细胞标志物糖蛋白激素受体5 mRNA表达与结直肠癌患者年龄、性别、肿瘤大小及部位、淋巴转移以及预后等临床病理特征的相关性。 结果与结论：趋化因子受体4、糖蛋白激素受体5 mRNA在结直肠癌组织中均有较高的表达率,但是趋化因子配体12 mRNA表达率降低。趋化因子受体4、干细胞标志物糖蛋白激素受体5 mRNA、趋化因子配体12 mRNA三者与结直肠癌患者的年龄、性别等患者临床特征无相关性,与结直肠癌的发病位置及其大小也无相关性,与结直肠癌组织是否淋巴转移具有相关性关,伴有淋巴转移的结直肠癌组织中干细胞标志物糖蛋白激素受体5 mRNA和趋化因子受体4的表达率更高,而趋化因子配体12 mRNA表达无显著变化;趋化因子受体4表达随肿瘤的恶性程度增高而增高;糖蛋白激素受体5表达于胃肠道肿瘤和脑肿瘤干细胞等表面,其表达随肿瘤的恶性程度增高而增高。提示结直肠癌组织中趋化因子受体4的表达增高,糖蛋白激素受体5基因表达增高,二者增高促进了结直肠癌组织生长及转移,糖蛋白激素受体5以及趋化因子配体12/趋化因子受体4轴的表达的调控,使其或将成为肿瘤诊断及治疗的重要新靶点。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

A novel antimetabolite, TAS-102 retains its effect on FU-related resistant cancer cells

TAS-102 is a new oral anti-tumor drug preparation, composed of a 1:0.5 mixture (on a molar basis) of alpha,alpha,alpha-tri-fluorothymidine (FTD) and thymidine phosphorylase inhibitor (TPI). TAS-102 is currently undergoing clinical trials, and has been demonstrated to have at least 2 mechanisms; inhibition of thymidylate synthase (TS) and incorporation into DNA. 5-FU is widely used in the treatment of solid tumor, but the inherent or acquired resistance of certain tumors to 5-FU therapy is a major clinical problem. In the present study, we investigated FTD in vitro and in vivo comparing with 5-FU and using FU-resistant cells. There was no relationship between FTD and 5-FU growth inhibition effect in vitro. A different sensitivity pattern was observed by the log-mean graph. We next investigated the anti-tumor activity of TAS-102 in a FU-resistant xenograft model. Comparative efficacy was observed between FU-resistant cell and its parent cell. We also studied the influence of TAS-102 on liver metastasis in a mouse model of human colorectal cancer, because liver metastasis of colorectal cancer is associated with patient survival. Human cancer DNA was detected by PCR, and TAS-102 markedly inhibited the number of liver metastasis. A novel angiogenic factor, platelet-derived endothelial cell growth factor (PD-ECGF), was shown to be identical to a previously characterized intracellular enzyme, thymidine phosphorylase, TAS-102 can be expected to have not only anti-tumor cytocidal effects but also antiangiogenesis activity and may inhibit liver metastasis. Our findings suggested that TAS-102 is a promising candidate for clinical use and can be expected to decrease minimal residual disease.     

Prognostic significance of the molecular markers in human gastrointestinal cancer

Matrix metalloproteinases have been implicated in tumor progression. Matrilysin is one of the matrix metalloproteinases and is frequently overexpressed in gastrointestinal cancers. The aim of this study was to assess the validity of matrilysin as a prognostic marker of colorectal cancers. Matrilysin expression was immunohistochemically analyzed using formalin-fixed, paraffin-embedded specimens from 113 colorectal cancer patients who had undergone curative surgery. The lumenal surface of neoplastic glands in the superficial layer was apically stained, while the cytoplasm of cancer cells at the invasive front was diffusely stained for matrilysin. Sections with immunostaining signals in more than 30% of carcinoma cells at the invasive front, which were observed in 47(42%) cases, were judged as being positive for matrilysin. Matrilysin positivity was significantly correlated with the depth of invasion, lymph node metastasis, lymphatic invasion, advanced Dukes' stage, and poor outcome. Patients with matrilysin-positive cancer had a significantly shorter overall survival time than those with matrilysin-negative cancer. For patients with intermediate invasive tumor(T2 or T3), only matrilysin was a significant prognostic variable for predicting overall survival in multivariate analysis. Matrilysin expression at the invasive front could be an important marker, predicting an unfavorable prognosis after surgical treatment in patients with colorectal cancer.     

高尔基体磷蛋白3在结直肠癌组织中表达及其对结直肠癌细胞增殖、侵袭和迁移的影响

目的 分析高尔基体磷蛋白3（GOLPH3）在结直肠癌组织中的表达情况,及其对结直肠癌细胞增殖、侵袭和迁移的影响。方法 收集术前未经放、化疗治疗,经手术切除患者的结直肠癌组织及其相应的癌旁组织（40例）;采用免疫组织化学法检测组织中GOLPH3的表达,分析GOLPH3与结直肠癌临床病理特征的关系;构建稳定沉默GOLPH3的结直肠癌细胞株,并设立阴性对照组和空白对照组,Western blotting法检测3组细胞中GOLPH3蛋白的表达,MTT法检测3组细胞的增殖活性,Transwell实验分别检测3组细胞的侵袭和迁移能力。结果 GOLPH3在癌组织中主要为阳性表达,且阳性表达率（65.0%）显著高于癌旁组织（35.0%）（P＜0.05）;GOLPH3在TNM分期为Ⅲ～Ⅳ期、浸润深度≥2 cm、中低分化及有淋巴结转移的癌组织中的阳性表达率明显高于TNM分期Ⅰ～Ⅱ期、浸润深度＜2 cm、高分化及无淋巴结转移的癌组织（P＜0.05）。与阴性对照组和空白对照组比,基因沉默组GOLPH3蛋白表达量、细胞增殖活性、侵袭能力及迁移能力均明显降低（P＜0.05）。结论 GOLPH3在结直肠癌组织中高表达,其表达与肿瘤分期、侵袭深度、分化程度及淋巴结转移有关,抑制GOLPH3基因表达可下调GOLPH3蛋白表达,抑制结直肠癌细胞增殖、侵袭和迁移。     

TS、DPD表达水平对5-FU疗效的预测价值

目的:探讨胸苷酸合成酶(TS)及二氢嘧啶脱氢酶(DPD)转录水平及其对5-氟脲嘧啶疗效的预测价值.方法:利用逆转录聚合酶链反应(RT-PCR)方法从胃肠癌细胞株中扩增胸苷酸合成酶及二氢嘧啶脱氢酶的DNA片段,并与内参照基因GAPDH进行比较.结果:对5-FU敏感的细胞株MKN45 TS和DPD的表达水平较低,而其他对5-FU不敏感的细胞株则无此现象.结论:TS及DPD mRNA水平对应用5-氟脲嘧啶治疗胃肠癌的疗效具有一定预测价值.     

FAT10基因表达与结直肠癌患者临床病理因素及预后的关系

目的:探讨结直肠癌(CCT)组织中FAT10(Diubiquitin,双泛素)的基因表达与其病理因素及预后的关系。方法:收集我院2000～2003年73例有5年以上完整随访资料的结直肠癌术后患者,采用RT-PCR方法检测FAT10 mRNA在CCT、癌旁组织(TCT)和正常结直肠组织(NCT)中的表达情况,并分析临床病理因素对FAT10阳性表达率和相对表达量的影响,以及FAT10表达与预后的关系。结果:FAT10 mRNA在CCT阳性表达率为52.24%(35/67),且在CCT的相对表达量(0.735±0.034)明显高于TCT(0.561±0.044)和NCT(0.397±0.028)(P<0.05);FAT10 mRNA的阳性表达率及相对表达量受临床进展、淋巴结转移和TNM分级的影响较明显(P<0.05),而患者年龄、肿瘤大小、肿瘤分化程度等对其无明显影响(P>0.05);Log-Rank检验显示CCT患者中,FAT10表达阳性者生存率明显低于FAT10表达阴性者(P<0.01)。结论:FAT10在CCT的表达状况与CCT转移及预后关系密切,可以作为反映生物学行为和判断预后的有效指标。     

结直肠腺癌中TKT的表达及临床意义

目的 探讨结直肠腺癌中转酮醇酶(transketolase,TKT)的表达及与临床病理特征的关系.方法 通过Oncomine和GEO数据库分析TKT基因在结直肠腺癌和癌旁组织中的表达.收集配对结直肠腺癌及癌旁正常肠黏膜组织,分别采用qRT-PCR、Western blot法检测TKT mRNA和蛋白表达,运用免疫组化法...     

Development of liver metastasis in colorectal carcinoma. With special reference to venous invasion and basement membrane laminin.

The development of hepatic metastases in 344 patients with colorectal carcinoma was examined for correlation with the presence of both venous invasion by the primary tumor and basement membranes in the tumor tissue. The former was detected by Victoria blue and hematoxylin-eosin staining and the latter by antilaminin antibody. A significant difference in the incidence of venous invasion was noted between patients with and those without liver metastasis at surgery. Basement membrane deposition was found in half of all cases of well differentiated adenocarcinoma, which was significantly high compared with other tumor types. This was more distinct in metastatic foci in the liver and lymph nodes than in the primary lesion, but less marked in intravascular tumor tissue. Basement membrane deposition was seen more frequently in Dukes' A tumors than in B tumors, although this was not statistically significant. No relationship was found between basement membrane laminin positivity and five-year survival, nor was there any correlation between the incidence of liver metastasis and tumor histologic type. Venous invasion was considered to be intimately related to the development of liver metastasis. Deposition of laminin-positive basement membrane was dependent on the grade of tumor differentiation, whereas it had no direct relation to the development of liver metastasis.     

p53和nm23蛋白表达与大肠癌浸润转移的关系

目的：探讨大肠癌组织中ｐ５３和ｎｍ２３蛋白表达与癌组织分化浸润转移的关系,以及它们之间的相关性。方法：应用免疫组织化学（ＳＡＢＣ）方法对４１例大肠癌组织中ｐ５３和ｎｍ２３蛋白表达情况进行检测。结果：大肠癌组织中ｐ５３和ｎｍ２３蛋白表达的阳性率分别为５８．５％和５３．７％,ｐ５３过表达、ｎｍ５３低表达与大肠癌浸润深度和淋巴转移具有相关性,ｐ５３过表达在低分化腺癌中明显高于高分化腺癌（Ｐ〈０．０１）;而ｎｍ２３高表达与大肠癌的分化程度无关（Ｐ〉０．０５）。ｎｍ２３低表达与ｐ５３高表达有相关性。结论：ｐ５３高表达、ｎｍ２３低表达在大肠癌的浸润转移中起重要作用,ｐ５３高表达与大肠癌组织分化程度有关,大肠癌组织浸润与淋巴转移可能与多基因异常改变有关。     

High extracellular matrix metalloproteinase inducer/CD147 expression is strongly and independently associated with poor prognosis in colorectal cancer

As in most solid tumors, colorectal cancer prognosis strongly depends on the extent of local invasion and lymph node and distant metastases. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a transmembrane glycoprotein that activates matrix metalloproteinases, a group of enzymes that play an important role in tumor invasion and metastasis formation. This study investigates the EMMPRIN expression in a large cohort of patients with colorectal cancer. Immunohistochemical analysis of tissue microarrays from 285 patients shows that increased EMMPRIN protein expression does not correlate with clinicopathologic parameters and is an independent prognostic factor of poor survival, with mean survival times of 103 months in EMMPRIN negative/low versus 57 months in EMMPRIN intermediate/high patients (P < .001). This pronounced association of increased EMMPRIN levels and--on average--a 45% reduction in overall survival could help improve the risk stratification in patients with colorectal cancer; moreover, the lack of correlations with classical measures of cancer invasion/spreading may suggest the relevance of alternative EMMPRIN pathways beyond matrix metalloproteinase activation.