Antisense oligodeoxynucleotide of glyceraldehyde-3-phosphate dehydrogenase gene inhibits cell proliferation and induces apoptosis in human cervical carcinoma cell lines
Antisense oligodeoxynucleotide of glyceraldehyde-3-phosphate dehydrogenase gene inhibits cell proliferation and induces apoptosis in human cervical carcinoma cell lines
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DOI：
10.1089/oli.1.1999.9.507
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发表时间：
1999-12-01
期刊：
影响因子：
--
通讯作者：
Namkoong, SE
中科院分区：
文献类型：
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作者：
Kim, JW;Kim, TE;Namkoong, SE
肿瘤细胞特征性地表现出糖酵解速率增加。在人类宫颈癌中发现了较高水平的 3-磷酸甘油醛脱氢酶 (GAPDH) mRNA。本研究旨在使用 GAPDH 反义寡脱氧核苷酸 (ODN) 硫代磷酸酯 (PS) 来评估人宫颈癌中 GAPDH 表达的改变如何影响生长抑制和诱导细胞凋亡。 Northern blot分析显示，与正常宫颈组织相比，三种宫颈癌细胞系（HeLa、CUMC-3和CUMC-6）的GAPDH基因表达水平显着升高，逆转录聚合酶链反应（RT-PCR）显示，10μM GAPDH反义ODN均抑制三种细胞系中GAPDH基因的表达，Western blot分析显示，GAPDH反义ODN处理后，GAPDH蛋白水平降低或缺失。在培养的宫颈癌细胞系中持续 12 天。暴露于 GAPDH 反义 ODN 的宫颈癌细胞系显示细胞增殖减少，并伴随着集落形成效率降低。 GAPDH 反义 ODN 对培养癌细胞的这种作用与细胞凋亡过程相关，包括 DNA 碎片增加。这些结果表明，未来使用针对宫颈癌特异性 GAPDH mRNA 的反义 ODN 进行基因治疗可能是另一种针对人类宫颈癌的治疗工具。
Tumor cells characteristically exhibit an increased rate of glycolysis. A higher level of glyeraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was found in human uterine cervical cancers. This study was designed using GAPDH antisense oligodeoxynucleotide (ODN) phosphorothioate (PS) to evaluate how alterations of GAPDH expression in human cervical carcinoma could influence growth inhibition and induction of apoptosis. Northern blot analyses revealed that the levels of GAPDH gene expression were strongly elevated in three cervical carcinoma cell lines (HeLa, CUMC-3, and CUMC-6) compared with normal cervical tissue, Reverse transcription-polymerase chain reaction (RT-PCR) showed that expression of the GAPDH gene was inhibited by 10 mu M of GAPDH antisense ODN in all three cell lines, Western blot analysis showed that the levels of GAPDH protein were decreased or absent after GAPDH antisense ODN treatment for 12 days in cultured cervical carcinoma cell lines. Cervical carcinoma cell lines exposed to GAPDH antisense ODN showed reduced cellular proliferation, which was accompanied by reduced colony-forming efficiency. This effect of GAPDH antisense ODN on cultured carcinoma cells was associated with the apoptotic process, including increased DNA fragmentation. These results suggest that future gene therapy using antisense ODN directed against cervical cancer-specific GAPDH mRNA might be another therapeutic tool against human uterine cervical carcinomas.