RNA干扰沉默α1，3岩藻糖转移酶-Ⅶ基因对人肝癌细胞增殖的影响
Effect of α1,3-fucosyltransferase-Ⅶ Gene Silence by RNA Interference on Cell Proliferation of Human Hepatocarcinoma Cells
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摘要: 目的：研究RNA干扰沉默α1,3 岩藻糖转移酶-Ⅶ （α1,3FucT-Ⅶ） 基因对人肝癌细胞增殖的影响。方法：设计合成α1,3FucT-Ⅶ特异性RNA干扰质粒，并用脂质体转染导入人肝癌H7721细胞。RT-PCR检测细胞中α1,3FucT-Ⅶ mRNA的表达。流式细胞术测定细胞表面唾液酸化的Lewis X抗原（SLex）的表达及细胞周期。细胞生长曲线通过MTT测定。p27Kip1的表达检测采用Western blot。结果：成功构建了α1,3FucT-Ⅶ特异的shRNA真核表达质粒载体。RNA干扰下调α1,3FucT-Ⅶ表达能抑制H7721细胞生长，表现为细胞生长速度明显较对照组细胞慢，S期细胞百分数明显降低，p27Kip1的表达增加。结论：α1,3FucT-Ⅶ可能通过调节p27Kip1的降解影响H7721细胞增殖。Abstract: Objective：To study the effect of α1,3 fucosyltransferase-Ⅶ （FucT-Ⅶ） gene silence on the hepatocarcinoma cell proliferation. Methods：α1,3FucT-Ⅶ specific RNA plasmid vector was constructed and then transfected into human H7721 hepatocarcinoma cell line with lipofectamine 2000. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the level of α1,3FucT-Ⅶ mRNA. Cell surface SLex and cell cycles were analyzed by flow-cytometry. Cell growth was detected by MTT method. The expression of p27 Kip1 was examined using western blot. Results：The specific shRNA targeted to α1，3FucT-Ⅶ gene could down regulate the expression of α1,3FucT-Ⅶ and inhibit the proliferation of H7721 cells. It was evidenced by decreasing the rate of cell growth and blocking the cells going from G1 to S stage, as well as increasing the expression of p27Kip1. Conclusion：α1，3FucT-Ⅶ may affect cell proliferation of H7721 cells by regulating the degradation of p27Kip1.
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