目的:探讨lgr5甲基化与人类卵巢癌临床病理之间的联系,以及其作为判定总的生存率的价值。方法:采用甲基化特异性聚合酶链式反应(methylation specific polymerase chain reaction,MSP)方法分析卵巢癌患者石蜡组织中lgr5启动子DNA甲基化...目的:探讨lgr5甲基化与人类卵巢癌临床病理之间的联系,以及其作为判定总的生存率的价值。方法:采用甲基化特异性聚合酶链式反应(methylation specific polymerase chain reaction,MSP)方法分析卵巢癌患者石蜡组织中lgr5启动子DNA甲基化情况。结果:lgr5在正常卵巢组织中几乎无甲基化改变,而在良性卵巢肿瘤中频率升高,上皮性卵巢癌组织中甲基化频率明显升高(P<0.01);lgr5甲基化与低分化上皮性卵巢癌(P<0.01)、肿瘤分期Ⅲ～Ⅳ(P<0.05)以及淋巴结转移(P<0.01)相关,但与组织类型关系不明显(P>0.05)。在70例上皮性卵巢癌患者组织中,lgr5甲基化频率为62.9%(44/70),并且5年总的生存率明显低于未甲基化的患者(P<0.05)。结论:lgr5甲基化与上皮性卵巢癌分化程度、肿瘤分期Ⅲ～Ⅳ以及淋巴结转移密切相关,同时也可以作为上皮性卵巢癌患者总的生存率的判定指标之一。展开更多
AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domain...AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domains), p16, RUNX3 (runt-related transcription factor 3), E-cadherin, hMLH1 (mutL homolog 1), ABCG2 (ATP-binding cassette, sub-family G, member 2) and BNIP3 (BCL2/adenovirus E1B 19 kDa interacting protein 3) were analyzed in 80 specimens of PF by quantitative methylation-specific polymerase chain reaction (PCR). Eighty patients were divided into 3 groups; Group A (n=35):the depth of cancer invasion was less than the muscularis propria; Group B (n=31): the depth of cancer invasion was beyond the muscularis propria. Both group A and B were diagnosed as no cancer cells in peritoneal cytology and histology; Group C (n=14): disseminated nodule was histologically diagnosed or cancer cells were cytologically defi ned in the peritoneal cavity.RESULTS: The positive rates of methylation in CHFR, E-cadherin and BNIP3 were significantly different among the 3 groups and increased in order of group A, B and C (0%,0% and 21% in CHFR, P<0.05; 20%, 45% and 50% in E-cadherin, P<0.05;26%,35% and 71% in BNIP3, P<0.05). In addition, the multigene methylation rate among CHFR, E-cadherin and BNIP3 was correlated with group A, B and C (9%,19% and 57%, P<0.001). Moreover, the prognosis was analyzed in group B, excluding 3 patients who underwent a non-curative resection. Two of the 5 patients with multigene methylation showed peritoneal recurrence after surgery, while those without or with a single gene methylation did not experience recurrence (P<0.05).CONCLUSION: This study suggested that gene methylation in the PF could detect occult neoplastic cells in the peritoneum and might be a risk factor for peritoneal metastasis.展开更多
AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected ...AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction （MSP） in four esophageal cancer cell lines （KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines）, nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine （5-aza-dc）, a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS： MSP results were as follows： loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation （com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells）; all nine cases of normal esophageal mucosa were unmethylated; and 54/72 （75.00%） samples from patients with esophageal can- cer were methylated, and 25/72 （34.70%） matched adjacent tissues were methylated （75.00% vs 34,70%, z2 = 23.5840, P = 0.000）. mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue （Z2= 9.143, P = 0.002）. XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation （Fisher＇s exact test, P = 0.004）. Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION： XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.展开更多
AIM: To explore the association of methylation of the CpG island in the promotor of the p16 tumor suppressor gene with the clinicopathological characteristics of the colorectal cancers. METHODS: Methylation-specific P...AIM: To explore the association of methylation of the CpG island in the promotor of the p16 tumor suppressor gene with the clinicopathological characteristics of the colorectal cancers. METHODS: Methylation-specific PCR (MSP) was used to detect p16 methylation of 62 sporadic colorectal cancer specimens. RESULTS: p16 methylation was detected in 42% of the tumors.Dukes'staging was associated with p16 methylation status.p16 methylation occurred more frequently in Dukes'C and D patients (75.9%) than in Dukes'A and B patients (12.1%). CONCLUSION: p16 methylation plays a role in the carcinogenesis of a subset of colorectal cancer, and it might be linked to poor prognosis.展开更多
FHIT基因是一种与凋亡有关的肿瘤抑制基因,广泛存在于各种细胞凋亡系统中,在多种肿瘤及细胞系中失活,其启动子部位CpG岛甲基化是FHIT基因失活的主要机制之一。本研究应用甲基化特异性PCR(Methylation specific PCR,MSP)对不同宫颈鳞癌...FHIT基因是一种与凋亡有关的肿瘤抑制基因,广泛存在于各种细胞凋亡系统中,在多种肿瘤及细胞系中失活,其启动子部位CpG岛甲基化是FHIT基因失活的主要机制之一。本研究应用甲基化特异性PCR(Methylation specific PCR,MSP)对不同宫颈鳞癌细胞系FHIT基因启动子CpG岛甲基化状态进行检测,在分子水平上了解FHIT异常甲基化与宫颈癌的相关性,为宫颈癌去甲基化治疗提供依据。展开更多
文摘目的:探讨lgr5甲基化与人类卵巢癌临床病理之间的联系,以及其作为判定总的生存率的价值。方法:采用甲基化特异性聚合酶链式反应(methylation specific polymerase chain reaction,MSP)方法分析卵巢癌患者石蜡组织中lgr5启动子DNA甲基化情况。结果:lgr5在正常卵巢组织中几乎无甲基化改变,而在良性卵巢肿瘤中频率升高,上皮性卵巢癌组织中甲基化频率明显升高(P<0.01);lgr5甲基化与低分化上皮性卵巢癌(P<0.01)、肿瘤分期Ⅲ～Ⅳ(P<0.05)以及淋巴结转移(P<0.01)相关,但与组织类型关系不明显(P>0.05)。在70例上皮性卵巢癌患者组织中,lgr5甲基化频率为62.9%(44/70),并且5年总的生存率明显低于未甲基化的患者(P<0.05)。结论:lgr5甲基化与上皮性卵巢癌分化程度、肿瘤分期Ⅲ～Ⅳ以及淋巴结转移密切相关,同时也可以作为上皮性卵巢癌患者总的生存率的判定指标之一。
基金Supported by Fund from the Department of Surgery,Saga University Faculty of Medicine
文摘AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domains), p16, RUNX3 (runt-related transcription factor 3), E-cadherin, hMLH1 (mutL homolog 1), ABCG2 (ATP-binding cassette, sub-family G, member 2) and BNIP3 (BCL2/adenovirus E1B 19 kDa interacting protein 3) were analyzed in 80 specimens of PF by quantitative methylation-specific polymerase chain reaction (PCR). Eighty patients were divided into 3 groups; Group A (n=35):the depth of cancer invasion was less than the muscularis propria; Group B (n=31): the depth of cancer invasion was beyond the muscularis propria. Both group A and B were diagnosed as no cancer cells in peritoneal cytology and histology; Group C (n=14): disseminated nodule was histologically diagnosed or cancer cells were cytologically defi ned in the peritoneal cavity.RESULTS: The positive rates of methylation in CHFR, E-cadherin and BNIP3 were significantly different among the 3 groups and increased in order of group A, B and C (0%,0% and 21% in CHFR, P<0.05; 20%, 45% and 50% in E-cadherin, P<0.05;26%,35% and 71% in BNIP3, P<0.05). In addition, the multigene methylation rate among CHFR, E-cadherin and BNIP3 was correlated with group A, B and C (9%,19% and 57%, P<0.001). Moreover, the prognosis was analyzed in group B, excluding 3 patients who underwent a non-curative resection. Two of the 5 patients with multigene methylation showed peritoneal recurrence after surgery, while those without or with a single gene methylation did not experience recurrence (P<0.05).CONCLUSION: This study suggested that gene methylation in the PF could detect occult neoplastic cells in the peritoneum and might be a risk factor for peritoneal metastasis.
基金Supported by Grants from the National Basic Research Program(973 Program), No. 2012CB934002, 2010CB912802National Key Scientific Instrument Special Programme of China, No.2011YQ03013405National Science Foundation of China,No. 81121004, 81071953 and 81161120432
文摘AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction （MSP） in four esophageal cancer cell lines （KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines）, nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine （5-aza-dc）, a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS： MSP results were as follows： loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation （com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells）; all nine cases of normal esophageal mucosa were unmethylated; and 54/72 （75.00%） samples from patients with esophageal can- cer were methylated, and 25/72 （34.70%） matched adjacent tissues were methylated （75.00% vs 34,70%, z2 = 23.5840, P = 0.000）. mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue （Z2= 9.143, P = 0.002）. XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation （Fisher＇s exact test, P = 0.004）. Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION： XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.
基金Supported by the grants from Mjnistry of Public Health of China,No.98-1-303The Educational Committee of Shanghai,No.2000B02.
文摘AIM: To explore the association of methylation of the CpG island in the promotor of the p16 tumor suppressor gene with the clinicopathological characteristics of the colorectal cancers. METHODS: Methylation-specific PCR (MSP) was used to detect p16 methylation of 62 sporadic colorectal cancer specimens. RESULTS: p16 methylation was detected in 42% of the tumors.Dukes'staging was associated with p16 methylation status.p16 methylation occurred more frequently in Dukes'C and D patients (75.9%) than in Dukes'A and B patients (12.1%). CONCLUSION: p16 methylation plays a role in the carcinogenesis of a subset of colorectal cancer, and it might be linked to poor prognosis.
文摘FHIT基因是一种与凋亡有关的肿瘤抑制基因,广泛存在于各种细胞凋亡系统中,在多种肿瘤及细胞系中失活,其启动子部位CpG岛甲基化是FHIT基因失活的主要机制之一。本研究应用甲基化特异性PCR(Methylation specific PCR,MSP)对不同宫颈鳞癌细胞系FHIT基因启动子CpG岛甲基化状态进行检测,在分子水平上了解FHIT异常甲基化与宫颈癌的相关性,为宫颈癌去甲基化治疗提供依据。
