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目的以表面增强激光解吸及电离飞行时间质谱(SELDI-TOF-MS)技术检测针对人类端粒酶RNA的反义寡核苷酸(hTR-ASODN)、针对hTR的正义寡核苷酸(hTR-SODN)、针对人类端粒酶活性催化亚单位的ASODN(hTERT-ASODN)、针对hTERT的SODN(hTERT-SODN)、没食子酰表没食子儿茶素(EGCG)、3′-叠氮-3′-脱氧胸苷(AZT)、全反式维甲酸(ATRA)和盐酸阿酶素(ADM)8种端粒酶抑制剂分别作用人肝癌细胞SMMC-7721后差异蛋白表达的情况。方法运用SELDI-TOF-MS技术结合配套的弱阳离子交换型芯片(WCX-2)检测8种端粒酶抑制剂分别作用于SMMC-7721细胞后分泌蛋白和胞浆蛋白表达的变化。对照组为未加任何处理因素的SMMC-7721细胞。结果WCX-2蛋白芯片检测发现,8种端粒酶抑制剂作用后细胞株的分泌蛋白和胞浆蛋白分别存在不同程度的表达差异及共同低表达或高表达的差异蛋白。所有差异蛋白分子量均小于10000Da。结论8种端粒酶抑制剂作用后SMMC-7721细胞存在特异性差异蛋白和共性变化的蛋白,这些蛋白可能与端粒酶活性的调控密切相关。
Objective To detect antisense oligonucleotides (hTR-ASODN) targeting human telomerase RNA, surface-enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF- SODN), ASODN (hTERT-ASODN) against human telomerase activity catalytic subunit, SODN (hTERT-SODN) against hTERT, gallic acid epigallocatechin (EGCG) AZT, ATRA and ADM were used to detect the differential proteins expression in human hepatoma SMMC-7721 cells. Methods The expression of secreted proteins and cytoplasmic proteins of 8 kinds of telomerase inhibitors were detected by SELDI-TOF-MS combined with weak cation exchange chip (WCX-2). The control group was SMMC-7721 cells without any treatment factors. Results The results of WCX-2 protein chip assay showed that there were different expression levels of the secreted proteins and cytoplasmic proteins of the cell lines with different levels of telomerase inhibitor and the differential proteins with low expression or high expression. All differential proteins have molecular weights less than 10000 Da. CONCLUSIONS: Eight kinds of telomerase inhibitors can differentiate SMMC-7721 cells with different proteins and their common proteins. These proteins may be closely related to the regulation of telomerase activity.