Demethylation of the γ-synuclein gene CpG island in colorectal cancer and its clinical significance

Objective To explore the relationship between γ-synuclein gene expression and CpG island demethylation in colorectal cancer (CRC), and the relationship between the demethylation and clinicopathological factors of CRC. Methods The expression of γ-synuclein mRNA was examined in 30 pairs of tumor tissues and tumor-matched non-neoplastic adjacent tissues (NNAT) by RT-PCR.CRC cell lines including COLO205, LoVo, and SW480 were used and treated with a demethylating agent, 5-aza-2'-deoxycytidine (5-aza-C). Before and after the treatment, the expression of γ-synuclein mRNA in the cells was determined by RT-PCR, and bisulfite sequencing PCR was also used to analyze methylation status of CpG island. The methylation status of γ-synuclein was then examined in 67 CRC samples and 30 NNAT samples by nested methylation-specific PCR (NMSP) and real time methylationspecific PCR (real-time MSP). The relationship between the demethylation of γ-synuclein in CRC and clinicopathological factors was analyzed. Results The mean γ-synuclein mRNA expression was 0.66±0.34 in CRC samples, which was much higher than 0.45±0.26 in NNAT samples (P=0.011). 5-aza-C could induce expression and demethylation of γ-synuclein in COLO205, LoVo and SW480 cells. Γ-Synuclein gene was demethylated in 80.0%(24/30) of the CRC samples and 50.0%(15/30) of the NNAT samples.The demethylated status of γ-synuclein was much higher in CRC samples than that in NNAT samples (P=0.030), and was significantly correlated with clinical stage, lymph node involvement, and distant metastasis of CRC (P＜0.05). Conclusion The upregulation of γ-synuclein expression in CRC is primarily attributed to the demethylation of CpG island, which may be used as a marker for prognosis.     

A differentially methylated region of the DAZ1 gene in spermatic and somatic cells

Aim:To investigate the methylation status of the deleted in azoospermia 1(DAZ1)gene promoter region in differentcell types.Methods:Using CpG island Searcher software,a CpG island was found in the promoter region of theDAZ1 gene.The methylation status of this region was analyzed in sperm and leukocytes by bisulfited sequencing.Results:The methylation status of the CpG island in the DAZ1 gene promoter region differed in leukocytes andsperm:it was methylated in leukocytes,but unmethylated in sperm.Conclusion:A differentially methylated region ofthe DAZ1 gene exists in spermatic and somatic cells,suggesting that methylation of this region may regulate DAZ1gene expression in different tissues.(Asian J Androl 2006 Jan;8:61-67)     

Aberrant methylation of the TDMR of the GTF2AIL promoter does not affect fertilisation rates via TESE in patients with hypospermatogenesis

Increasing evidence shows a relationship between epigenetic regulation and male infertility. The GTF2A1L gene promoter contains the DNA methylation site of a tissue-specific differentially methylated region （TDMR）. Eighty-six patients with non-obstructive azoospermia were assessed for the DNA methylation state of CpG islands in the GTF2AIL promoter using testicular genomic DNA. Based on histological criteria, 26 of the 86 patients had normal spermatogenesis （controls）, 17 had hypospermatogenesis and 26 had a Sertoli cell-only phenotype or tubular sclerosis. GTF2AIL TDMR methylation was significantly lower in testes DNA from control samples than from hypospermatogenic samples （P=0.029）. Patients with hypospermatogenesis were divided into two subgroups： high DNA methylation （HM, n=5） and low DNA methylation （LM, n= 12）. The GTF2AIL TDMR methylation rate differed significantly between the HM and LM groups （P=0.0019）, and GTF2A 1L expression was significantly higher among the LM than in the HM patients （P=0.023）. High TDMR methylation was correlated with low GTF2AIL gene expression levels. Both groups demonstrated relatively good outcomes with respect to sperm retrieval, fertilisation, pregnancy and childbirth rates. We observed that aberrant GTF2AIL gene expression was not correlated with fertilisation rates. The testicular sperm extraction （TESE） technique may be used to overcome male infertility due to aberrant TDMR methvlation.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

dsRNA介导的PTEN基因表达上调及其与启动子区甲基化的关系

目的 观察以启动子区非CpG岛序列为靶点的dsRNA促进PTEN基因的表达,并探讨RNAa现象对PTEN基因启动子区DNA甲基化的影响.方法 将与PTEN基因启动子区非CpG岛DNA序列互补的双链RNA分子(dsPTEN) 转染入A549和H292肺癌细胞中,采用real-time聚合酶链反应(PCR)检测PTEN的表达,筛选能够促进PTEN基因表达的有效dsRNA序列.通过甲基化特异性PCR(MSP)产物测序,检测dsRNA转染前后启动子区甲基化程度的变化.结果 经多条dsRNA分别转染肺癌细胞,证实针对PTEN基因-57到-38的序列(dsPTEN2-2)转染H292肺癌细胞后出现PTEN基因表达的明显上调,上调最高达5.1倍(与对照dsRNA比较,P<0.05),证实了RNAa现象的存在;使用5-Aza处理后的A549和H292细胞与空白组比较CpG点甲基化明显减少(5±3比10±4、6±3比9±3,P均<0.05),而转染PTEN2-2 dsRNA后的两个细胞株CpG点甲基化情况与未转染组比较无明显变化(9±2比10±4、9±3比9±3,P均>0.05).结论 dsRNA可以促进PTEN基因表达的上调,RNAa现象并不改变PTEN基因启动子区甲基化程度.

Abstract：

Objective To evaluate the phosphatase and tensin homolog deleted on chromosome ten (PTEN) gene activation induced by double-standed RNA (dsRNA) in lung cancer cell line and its correlation with CpG island methylation in the promoter region. Methods Specific dsRNA complementary to the non-CpG island sequence in the promoter of PTEN gene was designed and transfected into H292 and A549 cell lines, and the expression of PTEN mRNA was determined by real-time quantitative polymerase chain reaction (PCR). The methylation status of the CpG island in the promoter region was tested by DNA sequencing on the bisulfate modified DNA. Results By testing several dsRNA sequences targeting promoter region of PTEN, a dsRNA named PTEN2-2 (target on -57 to -38 of the PTEN) was identified, which could upregulate the PTEN expression by 5.1 times at most (P<0.05, controlled with the dsControl). Epigenetic analysis of PTEN promoter region confirmed DNA hypermethylation. PTEN2-2 dsRNA transfection turned on the expression of PTEN without affecting the methylation status of promoter DNA (9±2 vs 10±4, 9±3 vs 9±3, both P>0.05), as the DNA methyltransferase inhibitor did (5±3 vs 10±4, 6±3 vs 9±3, both P<0.05). Conclusion The expression of PTEN mRNA could be enhanced by inducing the dsRNA into the cells, but no evidence was found that dsRNA could affect the methylation status of the CpG island in the promoter region of this gene.     

Expressions and significances of Nek2 and β-catenin in breast invasive ductal carcinoma

Objective To investigate the expression and significance of NIMA related kinases 2 ( Nek2) and β-catenin (β-cat) in breast invasive ductal carcinoma (IDC) and concomitant ductal carcinoma in situ (DCIS). The roles of Nek2 and β-acat in the development and progression of breast cancer were explored. Methods The protein expression of Nek2 and β-cat in the tissues from 186 patients with IDC, 75 concomitant DCIS, 80 normal tissue adjacent to carcinoma and 40 fibroadenoma of breast was detected by using immunohistochemistry. Their relationship between clinicopathologic parameters was analyzed. Results There were correlations between the Nek2 expression in cytoplasm and grade(x2=8. 756;P<0.05) as well as tumor size (x2=6. 518;P<0.05) in IDC. The positive expression of Nek2 in nuclei was 32/186 and 15/75 in IDC and DCIS, respectively. There were correlations between the β-cat expression on the cytomembrane and grade (x2=45.493;P<0. 01) , TNM stage (x2=9. 510;P<0. 01), node status (Z=-2.035;P<0. 05) and estrogen receptor (ER) status (Z=-3. 004;P<0. 01). The β-cat expression on the cytoplasm was related with TNM stage (x2=8. 194;P<0. 05). The expression of Nek2 and β-cat had no correlation with clinicopathologic parameters in concomitant DCIS (P>0. 05), the Nek2 expression in cytoplasm had a correlation with the β-cat expression in cytoplasm (r=0. 226,P<0.05) in concomitant DCIS, and the same relationship could also be seen in IDC (r=0. 368,P<0.01). There was significant difference in the β-cat expression on the cytomembrane between IDC and concomitant DCIS (Z=-3. 804,P<0. 01). In the normal tissue adjacent to carcinoma and fibroadenoma, the Nek2 expression in cytoplasm and nuclei Was low or negative;the β-cat expression on the cytomembrane was high but that Was low in cytoplasm.Conclusion Centrosome regulatory factor Nek2 and β-cat can support a new way to explore the mechanisms of breast tumorigrenesis.And they may become a new antitumor drug target in the future.     

水通道蛋白1和3在结直肠癌组织中的表达及其意义

Objective To investigate the significance of aquaporin-1 (AQP-1) and aquaporin-3 (AQP -3) in the development of colorectal carcinoma. Methods The expression of AQP-1 and AQP-3 was investigated using immunohistochemical staining with Streptavidin Perdcidase in tissues from colorectal adenoma (CRA, n=25), colorectal cancer (CRC, n=50), and adjacent mucosa (CRT, n=50).Results The positive rate of AQP-1 was 64%(32/50) in CRC, significantly higher than that in CRT (38%, 19/50) and CRA(32%,8/25)(P＜0.05). The expression of AQP-1 was associated with depth of invasion and lymph node metastasis in CRC patients(P＜0.05). The positive rate of AQP-3 was 56% in CRT, 44% in CRA, and 52% in CRC. There were no significant differences(P＞0.05). The expression of AQP-3 was associated with age, tumor diameter, and depth of invasion(P＜0.05). No significant correlation between the expression of AQP-1 and AQP-3 in CRC was shown by Spearman correlation analysis(P＞0.05). Conclusions AQP-1 expression is increased in CRC while the expression of AQP-3 is not. There is no correlation between the expression of AQP-1 and AQP-3 in CRC.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

上皮钙黏素1基因启动子甲基化对结肠癌上皮钙黏素和β-连接素表达的影响

目的 探讨上皮钙黏素基因(CDH1)启动子甲基化与结肠癌上皮钙黏素(E-cadherin)及β-连接素(β-catenin)的表达及临床病理特征的关系.方法 采用甲基化特异性PCR技术检测68例结肠腺癌组织、癌旁组织及正常黏膜组织中CDH1基因启动子甲基化的状况.采用免疫组织化学法检测E-cadherin及β-catenin蛋白的表达.结果 癌旁组织及癌组织中CDH1启动子甲基化的阳性表达分别为32.4%(22/68)、57.4%(39/68),正常组织均为阴性表达(P<0.05).E-cadherin在正常组织、癌旁组织及腺癌组织中阳性表达率分别为92.6%、66.2%和44.1%.正常组织中β-catenin均表达于细胞膜上,无胞质和(或)胞核表达,而β-catenin在癌旁组织及癌组织中胞质和(或)胞核表达分别为29.4%和50.0%.CDH1基因启动子甲基化阳性率与E-cadherin表达则呈负相关(r=-0.312,P=0.01),与β-catenin胞质和(或)胞核表达呈正相关(r=0.309,P=0.018).CDH1基因启动子甲基化及E-cadherin、β-catenin的异常表达均与结肠癌分化程度及转移密切相关(P<0.05).结论 CDH1基因启动子甲基化可能是导致结肠癌E-cadherin与β-catenin异常表达及肿瘤侵袭性增强的重要原因.

Abstract：

Objective To investigate the relationship between methylation of the CDH1 gene promoter on the expression of E-cadherin and β-catenin, and to evaluate the correlation with clinicopathological characteristics of the colonic carcinoma. Methods Methylation specific PCR (MSP) was used to detect CDH1 gene promoter methylation in the cancer tissue, adjacent tissues and normal tissues in 68 patients. The expression of E-cadherin and β-catenin was determined by immunohistochemistry staining. Results The positive rate of CDH1 gene promoter methylation was 32.4% in adjacent tissues and 57.4% in cancer tissue, while no detectable methylation was found in all the normal tissues. The difference was statistically significant. The positive rate of E-cadherin was 92.6% in the normal tissues, 66.2% in the adjacent tissues and 44.1% in the cancer tissues. In all normal tissues, β-catenin was expressed only at the cellular membrane but not in the cytosol or nucleus, while the expression of β-catenin was present in the cytosol or nucleus in 29.4% of the adjacent tissues and 50.0% of the cancer tissues. The positive rate of CDH1 gene promoter methylation was negatively correlated with E-cadherin expression (r =-0.312,P =0.01) and positively correlated with β-catenin cytosolic/nucleus expression(r=0.309,P=0.018). The differentiation and metastasis of colonic carcinoma were associated with the aberrant expression of E-cadherin, β-catenin, and methylation of CDH1 promoter (P<0.05). Conclusion CDH1 gene promoter methylation may lead to aberrant expression of E-cadherin and β-catenin in colonic carcinoma, and may play an important role in promoting the invasion of tumor.     

ABCG2蛋白在肝癌中表达的临床意义

Objective To investigate the expression and clinical significance of ABCG2 protein in hepato-cellular carcinoma (HCC). Methods Specimens of HCC were collected at The First Aifiliated Hospital of Sun Yat-sen University from January 2005 to December 2006. The expression of ABCG2 protein in 165 samples of HCC tissue, 25 samples of normal liver tissue and 40 samples of cirrhotic liver tissue was detected using immunohisto-chemistry. The correlation between the expression of ABCG2 protein and clinicopathological characters was then analyzed. Enumeration data, survival rate and the difference between groups were analyzed with a chi-square test, the Kaplan-Meier method and Log-rank test, respectively. Results ABCG2 protein expression was weakly posi-tive in all normal and cirrhotic liver tissues. In HCC tissues, the expression of ABCG2 protein was strongly positive in 66 cases and weakly positive in 99 cases. The expression of ABCG2 protein was related to tumor diameter, tumor number, adjacent organ invasion and TNM stages (χ2 =8. 130, 14. 279, 4. 820, 21. 179, P <0. 05). Kaplan-Meier survival analysis revealed that patients with strongly positive ABCG2 protein had a significantly lower 3-year overall survival (24. 1%) compared with those with weakly positive ABCG2 protein (39. 4%) (χ2 = 15.716, P<0.05). Conclusions The expression level of ABCG2 protein is related to tumor invasiveness, TNM stage and prognosis. ABCG2 has the potential to become a new target for HCC treatment.     

胰腺癌组织中程序性细胞死亡因子4与细胞色素C的关系

Objective To study the relationship between the expression of cytochrome c ( Cyt c) and programmed cell death 4 (PDCD4) in pancreatic cancer, and investigate the pathway of PDCD4 inducing the apoptosis of pancreatic cancer cells. Methods Pancreatic cancer specimens from 69 patients who received pancreatic resection from 1990 to 2002 in First Affiliated Hospital of China Medical University were collected. The expression of Cyt c in the 69 paraffin specimens of pancreatic cancer was detected by immunohistochemistry, and the expression of Cyt c in 8 samples of cold-preserved fresh pancreatic cancer and normal pancreatic tissues were detected by Western blot. The expression of PDCD4 and Cyt c in pancreatic cancer was analyzed by paired t test and chi-square test. Results Compared with normal pancreatic tissues, the expression of Cyt c in pancreatic cancer was significantly decreased. The positive expression rate of Cyt c in 69 samples of pancreatic cancer was 41% (28/69). The expression of Cyt c was positive in most patients with positive expression of PDCD4, and the expression of PDCD4 was negative in most patients with negative expression of Cyt c. The expression of PDCD4 and Cyt c was closely correlated with each other (χ2= 10.52, P < 0.05). Conclusions There is a close relationship between the expression of PDCD4 and Cyt c in pancreatic cancer. PDCD4 may induce the apoptosis of pancreatic cancer cells through mitochondrial pathway.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.     

Hvin和Ki-67蛋白在胆管癌组织中表达的意义

Objective To investigate the relationship between Livin and Ki-67 proteins, and the expres-sion and clinical significance of Livin and Ki-67 proteins in cholangiocarcinoma. Methods Fifty-five samples of cholangiocarcinoma tissue were collected in Shengjing Hospital from January 2002 to December 2003. The expres-sion of Livin and Ki-67 proteins in the 55 samples of cholangiocarcinoma tissue and 12 samples of chronic cholan-gitis tissue were detected by immunohistochemical assay. The relationship between the expression of Livin and Ki-67 proteins and the clinicopathological parameters of cholangiocarcinoma was analyzed. The results were analyzed by Spearman rank correlation coefficient, chi-square test and t test. Results The positive expression rate of Livin protein in cholangiocarcinoma was 71% (39/55), which was significantly higher than 0 (0/12)in chronic cholan-girls tissue (χ2=20.361, P<0.01). The expression of Livin protein was influenced by the differentiation of cholangiocarcinoma and the lymph node metastasis (χ2=4.193, 4.245, P <0.05). The positive expression rate of Ki-67 protein was 96% (53/55) in cholangiocarcinoma. The label index of Ki-67 in patients in clinical stage Ⅰ,Ⅱ,Ⅲ,Ⅳ,were 22%±16%, 33%±12%, 43%±15%, and 49%±10%, respectively. There was signifi-cant difference in the label index of Ki-67 between patients in clinical stage Ⅰ and those in clinical stages Ⅱ, Ⅲ,Ⅳ(t=2.307, 2.871, 3.957, P<0.05). The label index of Ki-67 was 43%±13 % in patients with local lymph node metastasis, and 34%±16% in patients without local lymph node metastasis, with statistical difference between the 2 groups (t=2.334, P<0.05). The expression of Livin protein in cholangiocarcinoma was positively correlated with the label index of Ki-67 (r=0.502, P<0.01). Conclusions Livin protein plays an important role in the pathogenesis and development of eholangiocarcinoma, and it also has correlation with the proliferating activity of cholangiocarcinoma cells. Combined detection of the expression of Livin and Ki-67 proteins may be helpful in judging the malignancy of cholangiocarcinoma and determining the prognosis of patients.