E1B缺陷腺病毒对鼻咽癌CNE-2细胞杀伤作用及其机理研究

Growth Inhibition and Mechanisms of E1B-Deleted Adenovirus on Nasopharyngeal Carcinoma CNE-2 Cells

目的 :研究E1B缺陷腺病毒dl15 2 0对鼻咽癌CNE 2细胞的杀伤作用及其作用机理。方法 :用MTT法和细胞病变试验 (CPE)测量dl15 2 0在体外对CNE 2细胞的抑制和杀伤作用 ,并通过测定病毒在CNE 2细胞中的复制产量和DAPI染色试验探讨其杀伤机理 ;瘤内注射dl15 2 0 ,观察其对CNE 2裸鼠移植瘤的治疗效果 ,用免疫织化方法检测肿瘤组织中病毒的复制。结果 :体外实验结果显示 :dl15 2 0能在CNE 2细胞中复制 ,导致明显的细胞病变 ,显著抑制CNE 2细胞的生长 ,在病变细胞中可见明显的核膨胀 ;瘤内注射dl15 2 0能明显抑制裸鼠移植瘤的生长 ,在肿瘤组织中可检测到病毒复制。结论 :dl15 2 0能在CNE 2细胞中复制并使之裂解 ,从而在体内外有效地抑制和杀伤CNE 2细胞。

Objective: To explore antitumoral efficiency and mechanism of dl1520, an E1B-deleted adenovirus against nasopharyngeal carcinoma CNE-2 cells. Methods: Growth inhibition of dl1520 on CNE-2 cells was examined with MTT colorimetric assay and cytopathic effect (CPE) assay. The propagation of dl1520 in CNE-2 cells was measured with plaque assay on 293 cells, and changes of nuclei in CNE-2 cells infected with viruses were observed under a fluorescence microscope after DAPI staining. For in vivo study, athymic mice bearing CNE-2 nasopharyngeal carcinoma xenografts were administered intratumorally with dl1520, tumor growth was monitored twice a week, and virus replication in tumor tissues was examined by immunohistrochemistry. Results: in vitro, dl1520 replicated in CNE-2 cells and induced obvous CPE, so inhibited effectively the growth of CNE-2 cells. Distinct nucleus expansion, but not the characteristics of apoptosis, was found in CNE-2 cells infected with dl1520. Compared with control, dl1520 inhibited the growth of tumor xenografts in vivo, and viral replication was found on a large scale in tumors treated with dl1520. Conclusions: The E1B-deleted adenovirus dl1520 replicated effectively in CNE-2 cells and inhibited the growth of CNE-2 cells both in vitro and in vivo.