金喜素对原代培养的人食管癌细胞的抗癌作用.docVIP

金喜素对原代培养的人食管癌细胞的抗癌作用 作者：姜涛，程庆书， 张涛，李小飞，朱以芳 【关键词】 托泊替坎 　　Antitumor effects of Topotecan on primary cultured human esophagus cancer cells 　　【Abstract】 AIM: To study the effect of Topotecan on the proliferation and apoptosis of primary cultured esophagus cancer cells and its possible mechanism. METHODS: Primary cultured esophagus cells were treated with topotecan of different concentrations for 48 h. MTT assay was used to detect the viability of cells. Morphological changes of apoptotic cells were studied by acridine orange staining and the apoptotic rates were detected by flow cytometer. Caspase3 activity was measured by colorimetric assay. RESULTS: After treatment with topotecan for 48 h, the cell proliferation was significantly inhibited in a dosedependent manner. Typical morphological changes of apoptotic cells could be detected by acridine orange staining in cells treated with topotecan with an apoptotic rate of (334?8)% but few apoptotic cells could be detected in cells without topotecan incubation. Topotecan markedly increased the caspase3 activity of esophagus cancer cells after treatment for 48 h. CONCLUSION: Topotecan can inhibit the proliferation of primary cultured esophagus cancer cells and induce apoptosis through the activation of caspase3. 　　【Keywords】 topotecan; esophageal neoplasms;cells,cultured;cell division; apoptosis; caspase3 　　【摘要】目的： 研究金喜素（Topotecan）对体外原代培养的人食管癌细胞增殖与凋亡作用的影响及可能的机制. 方法： 原代培养人食管癌细胞，MTT法检测金喜素对食管癌细胞增殖作用的影响；吖啶橙染色观察凋亡细胞形态学变化；流式细胞仪定量检测金喜素处理细胞后细胞凋亡情况；底物裂解法检测金喜素对食管癌细胞半胱天冬酶3（caspase3）活性的影响. 结果： 金喜素处理细胞48 h后，可明显抑制癌细胞增殖，并呈浓度依赖性；金喜素处理的食管癌细胞经吖啶橙染色可检测到大量凋亡细胞，流式细胞仪检测其凋亡率为(334±68)%，而未经金喜素处理组几乎未检测到凋亡细胞；金喜素处理食管癌细胞后其Caspase3活性较未处理组显著增高. 结论： 金喜素可抑制体外原代培养的食管癌细胞增殖，并通过促进Csapase3活性而诱导细胞凋亡. 　　【关键词】 托泊替坎；食管肿瘤；细胞，培养的；细胞分裂；脱噬作用；caspase3 　　0引言 　　金喜素即9二甲基氨基10羟基喜树碱，是近年来临床上用于治疗晚期肺癌的新型化疗药物，它能抑制拓扑异构酶Ⅰ（TOPOⅠ）的活性，使DNA断裂而发挥独特的抗癌作用［1］ . 已有研究表明金喜素对肺癌、乳腺癌、卵巢癌等均有治疗作用，但目前关于其对食管癌的治疗方面报道较少，其抗食管癌的作用及机制尚未明确. 由于肿瘤细胞原代培养基本保留了原有