bcl-2高表达细胞株的构建及其抗凋亡特性的观察

Construction of bcl-2 overexpression cell line and observation on its apoptosis resistance ability

目的：建立ｂｃｌ－２高表达的肿瘤细胞株，观察其抗凋亡特性。方法：利用基因转染技术，将人的ｂｃｌ－２基因ｃＤＮＡ克隆于逆转录病毒载体ｐＬＸＳＮ，通过ＰＡ３１７细胞包装成病毒后感染宫颈癌ＨｅＬａ细胞。结果和结论：经ＰＣＲ及Ｗｅｓｔｅｒｎ印迹证明转染成功，得到ｂｃｌ－２稳定高表达株Ｈｂｃ１７。用顺铂诱导凋亡，Ｈｂｃ１７比对照细胞株Ｈ１具有更明显的抗凋亡能力。

Objective: In order to study the apoptosis resistance mechanism of bcl-2, the bcl-2 overexpression cell line Hbc17 was constructed, and the characteristicg of apoptosis resistance ability of Hbc17 were observed. Methods: With the gene transfer technology, human bcl-2 cDNA was cloned to retroviral vector pLXSN, then transferred to viruspackaging cell line PA317. Virus supernatant was used to infect HeLa cells. Results and Conclusion: Success of transfection was demonstrated by PCR and Western blot, and bcl-2 stable and high expression cell line Hbc17 was obtained. It could be observed that Hbc17 held more apoptosis resistance ability than control cell line H1 did in cisplatininduced apoptosis. All these provided a basis for study of apoptosis. 