目的观察甲基化抑制剂5-氮杂-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5-Aza-CdR)对肺癌SPC-A1细胞增殖、细胞划痕和凋亡的影响,探讨抑癌基因分泌型卷曲相关蛋白1(secreted frizzled related protein 1,SFRP1)和O6-甲基鸟嘌呤-DNA甲基转...目的观察甲基化抑制剂5-氮杂-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5-Aza-CdR)对肺癌SPC-A1细胞增殖、细胞划痕和凋亡的影响,探讨抑癌基因分泌型卷曲相关蛋白1(secreted frizzled related protein 1,SFRP1)和O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methylguanine-DNA-methyltransferase,MGMT)基因启动子区DNA甲基化mRNA和蛋白在其中的表达和意义。方法CCK-8法检测不同浓度的5-Aza-CdR对人肺癌SPC-A1细胞增殖影响,划痕实验测定5-Aza-CdR对SPC-A1细胞迁移能力的影响,Hoechst 33258染色检测(0、3、10、30μmol·L^(-1))5-Aza-CdR处理肺癌SPC-A1细胞24h后细胞凋亡情况,RT-PCR、Western blot法检测SPC-A1细胞中SFRP1和MGMT的mRNA、蛋白表达。结果5-Aza-CdR可以浓度梯度的抑制肺癌SPC-A1细胞增殖,IC 50为21.2μmol·L^(-1);5-Aza-CdR(3、10、30μmol·L^(-1))作用48 h后,肺癌SPC-A1细胞划痕愈合分别为对照组的(92.4±2.6)%、(83.6±4.2)%、(76.7±4.5)%;5-Aza-CdR处理肺癌SPC-A1细胞后,出现典型的细胞凋亡形态学改变;不同浓度5-Aza-CdR(3、10、30μmol·L^(-1))处理SPC-A1细胞24 h后,SFRP1、MGMT mRNA和蛋白表达增加(P<0.05)。结论5-Aza-CdR可抑制肺癌SPC-A1细胞增殖,抑制肺癌细胞划痕修复和促进凋亡,可能与升高MGMT和SFRPl的甲基化表达有关。展开更多
摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-...摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-2012年11月在我院行冠脉造影检查确诊冠心病患者100例，并根据冠脉病变的支数分组，并人选同时期在我院行冠脉造影术明确无冠脉狭窄的非冠心病组（30例）。采用聚合酶链式反应（PCR）测定外周血单个核细胞中TIPE2的基因水平，采用双抗体夹心ELISA法测定血清中MCP-1和MMP-9水平。结果：①外周血单个核细胞TIPE2基因表达水平测定的结果：冠脉病变患者PBMC中TIPE2的基因表达水平明显低于非冠心病组，双支病变组低于单支病变组，多支病变组明显低于单支病变组和双支病变组，差异均具有统计学意义（P〈0．01）；②TIPE2基因表达水平与MCP-1、MMP-9水平呈负相关，相关系数分别为r=-0．560，r=-0．569，均P〈0．05）。结论：冠心病患者中TIPE2表达水平随冠脉病变血管支数的增加而降低，而且与MCP-1和MMP-9呈负相关。展开更多
Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarker...Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarkers for colorectal cancer（CRC） screening. Methods： Methylation-specific PCR（MSP） was performed to analyze methylation status of SFRP1 and SFRP2 in a blinded fashion in tumor tissues and in matched stool samples from 39 patients with primary CRC, 34 patients with adenomas, 17 patients with hyperplastic polyps and 20 endoscopically normal subjects as normal controls. Simultaneously we analyzed the correlation of hypermethylated SFRP1 and SFRP2 with the clinicopathological features of CRC. Results：Hypermethylated SFRP1 was detected in 92.3%, 76.5%, 47.1% of tissue samples and in 89.7%, 64.7%, 35.3% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Hypermethylated SFRP2 was detected in 87.2%, 67.6%, 35.3% of tissue samples and in 82.1%, 55.9%, 29.4% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Of these two genes, at least one hypermethylated was 94.9%, 82.4%, 52.9% in tissue samples and 92.3%,73.5%, 47.1% in matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. In contrast, no hypermethylated SFRP1 and SFRP2 were detected in mucosa tissues of normal controls, only 2 cases of fecal samples was detected with hypermethylated SFRP1 and another 1 case was detected with hypermethylated SFRP2. Moreover, no significant associations were observed between hypermethylated SFRP1,SFRP2 and clinicopathological features of CRC. Conclusion： Hypermethylation of SFRP1 and SFRP2 in feces are novel epigenetic biomarkers of CRC and carded high potential for the remote detection of CRC as non-invasive screening method, and combined analysis of hypermethylated SFRP1 and SFRP2 in fecal could further increase the detection rate of CRC and premalignant lesions.展开更多
文摘摘要目的：观察和分析冠心病患者中肿瘤坏死因子-α诱导蛋白-8样分子．2（tumornecrosisfactor．dinduced protein 8 like2，TIPE2）表达水平及其与单核细胞趋化因子-1（MCP-1）和基质金属蛋白酶-9（MMP-9）的相关性。方法：2010年12月-2012年11月在我院行冠脉造影检查确诊冠心病患者100例，并根据冠脉病变的支数分组，并人选同时期在我院行冠脉造影术明确无冠脉狭窄的非冠心病组（30例）。采用聚合酶链式反应（PCR）测定外周血单个核细胞中TIPE2的基因水平，采用双抗体夹心ELISA法测定血清中MCP-1和MMP-9水平。结果：①外周血单个核细胞TIPE2基因表达水平测定的结果：冠脉病变患者PBMC中TIPE2的基因表达水平明显低于非冠心病组，双支病变组低于单支病变组，多支病变组明显低于单支病变组和双支病变组，差异均具有统计学意义（P〈0．01）；②TIPE2基因表达水平与MCP-1、MMP-9水平呈负相关，相关系数分别为r=-0．560，r=-0．569，均P〈0．05）。结论：冠心病患者中TIPE2表达水平随冠脉病变血管支数的增加而降低，而且与MCP-1和MMP-9呈负相关。
基金supported by the grant from Programs of Science and Technology Commission Foundation of Jiangsu Province(NO.BS2005036)
文摘Objective：To investigate the feasibility of the combination of detecting hypermethylated secreted frizzled-related protein 1 （SFRP1） and secreted frizzled-related protein 2（SFRP2） in feces as a panel of biomarkers for colorectal cancer（CRC） screening. Methods： Methylation-specific PCR（MSP） was performed to analyze methylation status of SFRP1 and SFRP2 in a blinded fashion in tumor tissues and in matched stool samples from 39 patients with primary CRC, 34 patients with adenomas, 17 patients with hyperplastic polyps and 20 endoscopically normal subjects as normal controls. Simultaneously we analyzed the correlation of hypermethylated SFRP1 and SFRP2 with the clinicopathological features of CRC. Results：Hypermethylated SFRP1 was detected in 92.3%, 76.5%, 47.1% of tissue samples and in 89.7%, 64.7%, 35.3% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Hypermethylated SFRP2 was detected in 87.2%, 67.6%, 35.3% of tissue samples and in 82.1%, 55.9%, 29.4% of matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. Of these two genes, at least one hypermethylated was 94.9%, 82.4%, 52.9% in tissue samples and 92.3%,73.5%, 47.1% in matched fecal samples from CRC, adenoma and hyperplastic polyp, respectively. In contrast, no hypermethylated SFRP1 and SFRP2 were detected in mucosa tissues of normal controls, only 2 cases of fecal samples was detected with hypermethylated SFRP1 and another 1 case was detected with hypermethylated SFRP2. Moreover, no significant associations were observed between hypermethylated SFRP1,SFRP2 and clinicopathological features of CRC. Conclusion： Hypermethylation of SFRP1 and SFRP2 in feces are novel epigenetic biomarkers of CRC and carded high potential for the remote detection of CRC as non-invasive screening method, and combined analysis of hypermethylated SFRP1 and SFRP2 in fecal could further increase the detection rate of CRC and premalignant lesions.
