结肠癌组织中p15、MMP-2、MMP-9的表达及意义

目的 探讨p15、基质金属蛋白酶(MMP)-2、MMP-9在结肠癌中的表达及意义.方法 采用免疫组化SP法检测50例结肠癌组织、癌旁组织和正常切缘组织中的p15、MMP-2、MMP-9.结果 结肠癌组织、癌旁3 cm和正常切缘中,p15的阳性表达率分别为84%、94%、100%,MMP-2分别为76%、20%、0,MMP-9分别为82%、44%、0,三种组织中三项指标相比,P均<0.05.癌组织中三项指标的表达与肿瘤的分化程度、Dukes分期及有无淋巴结转移密切相关. 结论 p15的低表达和MMP-2、MMP-9的过表达在结肠癌的发生、发展及其浸润和转移中具有重要作用.     

Germline mutation analysis of hPMS2 gene in Chinese families with hereditary nonpolyposis colorectal cancer

AIM: To study the germline mutation of hPMS2 gene in 26 unrelated Chinese hereditary nonpolyposis colorectal cancer (HNPCC) probands and to fulfill the screening strategy for HNPCC in Chinese. METHODS: Genomic DNA was extracted from the peripheral blood. To avoid the interference of pseudogene in detection of the remaining 11 exons (exon 1-5, 9, 11-15), long-range polymerase chain reaction (PCR) was conducted to amplify the complete coding region of hPMS2 gene firstly. Then 1/8 of the PCR productswere used as template to amplify the individual exon respectively and DNA sequencing was done. Direct DNA sequencing of the conventional PCR products of exon 6, 7, 8 and 10 of hPMS2 gene was performed. The same analysis was made in 130 healthy persons without family histories of HNPCC to further investigate the pathological effects of the detected missense mutation. RESULTS: One HNPCC proband fulf illed Bethesda guidelines and was found to carry the germline mutation of hPMS2 gene, which has not been reported in Chinese HNPCC families. It was a missense mutation at c.1532C>T of exon 11. It was detected in three controls as well with an occurrence rate of 2.3% (3/130). Since it could not be found in the PMS2-single nucleotide polymorphism (SNP) database, this missense mutation is a new SNP unreported up to date. Meanwhile, 260 reported SNPs of hPMS2 gene were detected in the 26 HNPCC probands. The 2nd and 5th exons were probably the hot SNP regions of hPMS2 gene in Chinese HNPCC families involving 53.1% of all reported SNP. CONCLUSION: The germline mutation of hPMS2 gene may be rare in Chinese HNPCC families. The 2nd and 5th exons are hot SNP regions of hPMS2 gene.     

Clinical significance of human kallikrein 10 gene expression in colorectal cancer and gastric cancer

BACKGROUND AND AIM: Recent evidence suggests that the human kallikrein 10 (KLK10) gene is differentially regulated in endocrine-related tumors and has potential as diagnostic and/or prognostic marker; however, KLK10 expression has never been investigated in gastrointestinal cancers. The aims of this study were to demonstrate expression and single nucleotide polymorphisms of KLK10 in colorectal cancer (CRC) and gastric cancer (GC), and to correlate the relative KLK10 expression level with clinicopathological factors of CRC and GC. METHODS: Between March 2004 and January 2005, 63 patients with histologically confirmed CRC and 36 with GC were recruited into the study. Using quantitative real-time (qRT) RT-PCR and Western blot, KLK10 expression in tumor and non-tumor colorectal and gastric tissues was determined at the mRNA and protein levels. KLK10 protein was localized by immunohistochemistry. The KLK10 genomic DNA from 16 cases of paired normal/cancerous colorectal tissues was PCR-amplified and examined for single nucleotide polymorphisms by direct sequencing. RESULTS: KLK10 mRNA expression was detected by qRT in 61 of 63 (96.8%) CRC specimens and in all GC specimens. KLK10 expression was much higher in tumor tissue than in the corresponding normal mucosal tissue at the mRNA and protein levels (P<0.01). The KLK10 mRNA expression level significantly correlated with lymphatic invasion (P=0.034) and clinical stage of CRC (P=0.025). The KLK10 mRNA expression level significantly correlated with the depth of GC invasion (P=0.018), clinical stage (P=0.045), patient sex (P=0.027) and Lauren type of gastric cancer (P=0.028). No mutations or polymorphisms were detected in exon 1, 2 and 5 of KLK10 gene in CRC. Single nucleotide polymorphisms were identified in codon 50 of exon 3, GCC (alanine) to TCC (serine). The genetic changes of exon 4 were located at codon 106 [GGC (glysine) to GGA (glysine)], codon 112 [ACG (threonine) to ACC (threonine)], codon 141 [CTA (leucine) to CTG (leucine)], and codon 149 [CCG (proline) to CTG (leucine)]. All were identical in tumor and corresponding normal tissue DNA from the same individuals. CONCLUSION: KLK10 expression is up-regulated in CRC and GC and higher expression of KLK10 closely correlates with advanced disease stage, which predicts a poorer prognosis; however, further follow-up study is needed.     

Early postnatal dexamethasone influences matrix metalloproteinase-2 and -9, and their tissue inhibitors in the developing rat lung

In order to test the hypothesis that early postnatal exposure to dexamethasone (Dex) influences matrix metalloproteinases (MMP)-2 and -9, as well as their tissue inhibitors (TIMP-1 and -2) in the developing rat lung, newborn rats (3 litters/group) were treated with low Dex (0.1 mg/kg/day, IM), high Dex (0.5 mg/kg/day), or equivalent volumes of saline at 5 days postnatal age (P5), P6, and P7. Lung weight and lung MMP and TIMP levels were determined at sacrifice (7 days postinjection, P14; at weaning, P21; and at adolescence, P45, n = 10/group and time). Dex did not adversely affect lung weight or lung MMP-2 levels, which peaked in all groups at P21 and then fell by P45. In contrast, Dex decreased TIMP-2 at all time intervals, but achieved statistical significance only at P45. An imbalance in MMP-2/TIMP-2 ratio was noted at P21, with elevations occurring in the low and high Dex-treated groups. Lung MMP-9 levels remained comparable with controls during low Dex treatment. However, high Dex exposure resulted in elevated lung MMP-9 levels at P21 and P45. Lung TIMP-1 levels increased only with high Dex exposure at P14 and P21, whereas the lung MMP-9/TIMP-1 ratio was elevated at P21 in the high Dex group, and at P45 in both Dex-treated groups. These data provide evidence that early postnatal dexamethasone results in an imbalance between gelatinase-A and -B, and their tissue inhibitors in the developing rat lung. These changes may be responsible, in part, for some of the known maturational effects of steroids on lung structure in the newborn.     

A variant in microRNA-196a2 is not associated with susceptibility to and progression of colorectal cancer in Chinese

Background: MicroRNAs (miRNAs) are small non‐coding RNAs with regulatory functions as tumour suppressors and oncogenes. Although single nucleotide polymorphism (SNP) in miRNA regions have been reported to be rare and unlikely to be functionally important, recent evidence suggested that rs11614913 SNP in miR‐196a2 was associated with the susceptibility of lung cancer, breast cancer, congenital heart disease and shortened survival time of non‐small‐cell lung cancer. Aims: The aim of this study was to investigate the association between this genetic variant and the risk and/or progression of colorectal cancer (CRC). Methods: A total of 126 CRC patients and 407 healthy controls was periodically enrolled. DNA was extracted from blood specimens, and miR‐196a2 polymorphism was genotyped by polymerase chain reaction–ligation detection reaction (PCR–LDR). Results: Although the frequency of CC homozygotes or miR‐196a2C allele‐containing genotypes (CT and CC) was lower in CRC patients than in the healthy controls, no significant association between miR‐196a2 polymorphism and the risk of CRC was found. The frequency of the ‘C’ allele in CRC patients was also not significantly lower than in healthy controls. In a subsequent analysis of the association between this polymorphism and the progression of CRC, there was still no significant difference in both genotype and allelic frequency. Conclusions: Our results suggest that miR‐196a2 polymorphism is not associated with both an increased risk and progression of CRC in Chinese.     

Influence of cancer-related gene polymorphisms on clinicopathological features in colorectal cancer

Background and Aim:  Single nucleotide polymorphisms (SNP) are shown to be related with cancer incidence. It has been reported that CCND1 , p21 ^cip1 DCC , MTHFR, and EXO1 are related with the risk of malignant neoplasm, but few studies have mentioned the prognosis of the patients. We investigated the SNP of patients and related this to clinicopathological features, including survival rate.
Method:  DNA from the tissues of primary colorectal cancer was obtained from surgical resections of 114 patients (68 males and 46 females, 29–83 years). The CCND1 polymorphism was analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and those of other genes were investigated by the TaqMan method. The polymorphisms obtained were statistically analyzed for the relationship with clinicopathological features.
Results:  The CG  +  GG allele was more invasive than the CC allele in histological tumor depth in the DCC codon 201 ( P  = 0.0086). The 677TT allele in MTHFR had a larger tumor size than the 677CC allele ( P  = 0.028). In EXO1 P757L polymorphism, patients with the TT allele had a statistically reduced survival rate compared with the other alleles. In CCND1 polymorphisms, we found no statistical significance in clinicopathological features.
Conclusions:  From these preliminary data, these polymorphisms would be candidates predicting the clinicopathological features of colorectal cancer, but further more systematic gene analyses are warranted.     

Contents of tissue CEA and CA19-9 in colonic polyp and colorectal cancer,and their clinical significance

In precancerous states or early cancer, the serum levels of tumor markers are almost not detectable. Therefore, the tissue contents of CEA and CA19-9 were measured in 48 colonic polyps, 8 colorectal cancers and 5 normal colonie mucosa. These tissue specimens were obtained by endoscopie polypectomy, surgery or autopsy, and homogenated in normal saline (10 ml/wet g of tissue). After centrifugation, the supernatant was assayed by enzyme or radioimmunoassay. There was no correlation between serum levels and tissue contents of CEA or CA19-9 in colonie adenomas and colorectal cancers. The mean contents of tissue CEA and CA19-9 in colonie polyp and colorectal cancer were significantly higher than normal colonie mucosa, and the highest contents of CEA and CA19-9 were found in colorectal cancer. The contents of tissue CEA and CA19-9 in cancerous regions were markedly increased as compared with noncancerous regions. In adenomas, there was a relationship between the degree of histological dysplasia and the tissue content of CEA. Relationships were also found between macroscopic findings and tissue tumor markers in adenomas. These results suggest the possibility that the measurement of tissue tumor markers may be useful for borderline colonie lesions.     

结肠癌组织中MMP-7、MMP-9的表达及意义

目的观察结肠癌组织中基质金属蛋白酶（MMP）-7、MMP-9的表达,探讨其在结肠癌浸润、转移中的作用。方法采用免疫组化SP法检测50例结肠癌及其切缘组织中MMP-7、MMP-9的表达。结果肿瘤组织中MMP-7、MMP-9阳性表达率分别为68．00％和82．00％,标本切缘组织中分别为0和24．00％,两者相比,P均〈0．05,MMP-7、MMP-9在有淋巴结转移者中的阳性表达率为86．96％和91．30％,显著高于无淋巴结转移者的51．85％和74．07％,P均〈0．05;MMP-7、MMP-9在结肠癌组织中的表达呈正相关,r=0．84,P〈0．05。结论MMP-7、MMP-9高表达可能与结肠癌的浸润、转移有关。     

检测病变组织中CD146、MMP-9及TIMP-2对完全性葡萄胎恶变的预测价值

目的探讨黏附分子CD146、基质金属蛋白酶9（MMP-9）及基质金属蛋白酶抑制剂2（TIMP-2）的表达对完全性葡萄胎恶变的预测价值。方法采用免疫组化二步法检测80例完全性葡萄胎（恶变组36例,未恶变组44例）组织中CD146、MMP-9及TIMP-2的表达,分析其对完全性葡萄胎恶变的预测价值。结果完全性葡萄胎恶变组中CD146、MMP-9的表达明显高于未恶变组,P均〈0.01;TIMP-2在两组中的表达相近（P〉0.05）。联合检测CD146、MMP-9的表达对完全性葡萄胎恶变的阳性预测值和阴性预测值分别为84.1%和72.2%。结论CD146和MMP-9对完全性葡萄胎恶变的预测有一定价值,TIMP-2无预测价值。     

基质金属蛋白酶/基质金属蛋白酶组织抑制剂与心房颤动

基质金属蛋白酶是一个细胞外基质降解酶家族,基质金属蛋白酶组织抑制剂是其内源性抑制物.二者平衡在细胞外基质合成及降解过程中起重要作用,与心房颤动过程中心房结构重塑,即心房纤维化及心房扩大相关.目前研究显示基质金属蛋白酶/基质金属蛋白酶组织抑制剂失衡与心房颤动的发生、发展及复发相关,抑制基质金属蛋白酶可改善心房重塑并阻止心房颤动进展.恢复基质金属蛋白酶/基质金属蛋白酶组织抑制剂平衡可能成为治疗心房颤动的新型治疗途径.     

辛伐他汀对动脉粥样硬化斑块中MMPs及TIMPs表达的影响

目的 :观察辛伐他汀对动脉粥样硬化斑块中基质金属蛋白酶 （MMPs）及其组织抑制因子 （TIMPs）表达的影响。方法 :新西兰大白兔 48只随机分为试药组和对照组 ,用 PTCA球囊导管拉伤腹主动脉 ,高脂饮食喂养 4周后 ,试药组 （2 4只 ）给予辛伐他汀 5m g· kg- 1· d- 1,对照组 （2 4只 ）以淀粉为对照 ;每组按实验终点 （给药后 2、4和 8周 ）随机分为 3个亚组 ,继续高脂饮食喂养至实验结束。动物高脂喂养前、每周和处死前取静脉血 2 ml检测血脂变化 ,取拉伤段动脉用于总 RNA提取和组织病理检测。结果 :高脂饮食喂养 1周后 ,2组动物血脂水平在正常的 5～10倍 ,组间无显著差异 （P>0 .0 5） ;用药后 2周 ,试药组血脂明显低于对照组 ;用药后 4周 ,试药组血管组织MMP1,2低于对照组 ,TIMP1/ MMP1明显大于对照组 （P <0 .0 5,P<0 .0 1） ,TIMPs表达两组无显著差异 （P >0 .0 5）。结论 :辛伐他汀通过选择性地改变局部 MMPs和 TIMPs表达 ,减少基质分解破坏 ,增加局部斑块的稳定性     

Impact of MTHFR gene C677T polymorphism on Bcl-2 gene methylation and protein expression in colorectal cancer

Abstract

Objective. To investigate the impact of MTHFR C677T polymorphism on Bcl-2 gene promoter CpG island (CGI) methylation and Bcl-2 protein expression. Material and methods. MTHFR polymorphisms of 86 sporadic colorectal cancer (CRC) patients and 100 healthy volunteers were analyzed by PCR-based restriction fragment length polymorphism, and Bcl-2 promoter CGI methylation in 86 CRC tissues and 86 paired nonneoplastic adjacent tissues was determined by methylation-specific PCR. Bcl-2 oncoprotein expression in 70 CRC tissues and paired nonneoplastic adjacent tissues was detected by immunohistochemistry. Results. The frequency of MTHFR 677 T allele and combined variant genotypes (677CT + TT) in CRC patients was significantly higher than that in healthy controls (p = 0.023 and p = 0.035, respectively), and there is a significant association between 677TT or 677(CT + TT) genotypes and CRC (OR = 2.534, p = 0.045 and OR = 1.888, p = 0.035, respectively). The frequency of methylated Bcl-2 promoter CGI in tumor tissues was significantly lower than that in nonneoplastic adjacent tissues (p = 0.014). The frequency of methylated Bcl-2 promoter CGI in CRC tissues of the individuals with CC genotype was significantly higher than that of those with CT/TT genotypes (p = 0.018), there was significant distribution difference of C and T alleles between individuals with methylated and unmethylated Bcl-2 promoter CGI in colorectal cancer tissues (p = 0.023). Bcl-2 promoter hypomethylation was significantly correlated with Bcl-2 oncoprotein expression in colorectal cancer tissues (r = 0.558, p < 0.001). Conclusion. Bcl-2 promoter is hypomethylated in colorectal cancer tissue, and there is a significant correlation between MTHFR 677 TT or CT/TT genotypes and CRC or Bcl-2 promoter CGI methylation/oncoprotein expression in CRC.     

Differences in plasma gastrin,CEA, and CA 19-9 concentration in patients with proximal and distal colorectal cancer

Summary Aim. We investigated whether there are differences in plasma gastrin, as compared with carcinoembryonic antigen (CEA) and cancer antigen (CA) 19-9 between patients with proximal and distal colorectal cancer. Gastrin concentration has also been analyzed, dependent on the tumor stage, in order to evaluate the possible prognostic role of this measurement. Methods. In 50 patients with colon cancer-fasting gastrin, CA 19-9 and CEA levels were evaluated. Results. Mean plasma-gastrin level in patients with distal tumor yielded 105.31 ± 12.5 μU/L and was significantly higher than in patients with the proximal tumor site (42.2 ± 3.1 μU/L) as well as in controls (p<0.001). No significant difference was observed between mean plasma gastrin in patients with proximal tumors and the control group. The mean CEA plasma level was significantly higher (p<0.01) in patients with distal tumors (9.1 ± 1.1 ng/mL) than in those with proximal tumors (1.48 ± 0.1 ng/mL). Similarly, the mean CA 19-9 plasma level was significantly higher (p<0.01) in patients with distal tumor (19.9 ± 2.1 U/mL) than in those with proximal tumor: 1.8 ± 0.2 U/mL. The mean gastrin plasma, CA 19-9, and CEA level was significantly higher in group of Duke’s stage C and D as compared to A and B. Conclusion. We speculate that observed differences in gastrin concentration in patients with distal and proximal tumors may contribute to the distinct pathogenesis and biological properties of those cancers. The significance of gastrin as a marker for diagnostic or prognostic purposes in colorectal cancer requires further study.     

Twist1,MMP-2和MMP-9在结直肠癌组织中的表达及意义

目的:研究Twist1、MMP-2和MMP-9蛋白在结直肠癌组织中的表达及其相互关系.方法:建立组织微阵列平台,应用免疫组织化学方法检测92例结直肠癌组织Twist1、MMP-2和MMP-9蛋白的表达情况.结果:结直肠癌中Twist1的表达率为64.1%,MMP-2和MMP-9阳性率分别为66.3%和67.4%;Twi...