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目的：观察肿瘤选择性增殖腺病毒CNHK300对HER-2高表达乳腺癌细胞株的杀伤作用。方法：用TRAP- ELISA方法检测乳腺癌细胞株(MDA-MB-453、SK-BR-3)和正常成纤维细胞株MRC-5的端粒酶活性；进行病毒增殖实验和细胞生长抑制实验,并与野生型腺病毒wtAd5进行对比,验证CNHK300选择性复制和选择性杀伤能力；用细胞生长抑制实验进一步考察CNHK300与化疗药物紫杉醇联合应用对乳腺癌细胞的杀伤作用；用Western blot检测腺病毒E1A在细胞中的表达。结果：乳腺癌细胞株端粒酶均为阳性,而MRC-5正常成纤维细胞株端粒酶为阴性。CNHK300在乳腺癌细胞中的病毒增殖能力与野生型腺病毒相似,两者也具有相似的肿瘤细胞杀伤能力。在MRC-5正常成纤维细胞中,CNHK300病毒增殖能力较wtAd5明显减弱,同时对正常成纤维细胞的杀伤力明显减弱。CNHK300病毒和紫杉醇联合应用,较同样剂量的两者单独应用,细胞存活率明显下降。CNHK300病毒E1A可以选择性地在端粒酶阳性的乳腺癌细胞中表达,在端粒酶阴性的正常成纤维细胞中不表达。结论：肿瘤选择性增殖腺病毒CNHK300可选择性地在某些端粒酶阳性的乳腺癌细胞中复制,并产生溶瘤作用。与化疗药物紫杉醇联合应用可产生协同作用。
Objective: To observe the killing effect of tumor-selective adenovirus CNHK300 on HER-2 overexpressing breast cancer cell lines. Methods: The telomerase activity of breast cancer cell line (MDA-MB-453, SK-BR-3) and normal fibroblast cell line MRC-5 was detected by TRAP- ELISA. The virus proliferation test and cell growth inhibition test were performed. And compared with wild-type adenovirus wtAd5 to verify the selective replication and selective killing ability of CNHK300. The cytotoxicity of CNHK300 combined with paclitaxel on breast cancer cells was further investigated by cell growth inhibition assay. Western blot was used to detect the expression of E1A Expression in cells. Results: The telomerase activity of breast cancer cell lines was positive, while the telomerase of MRC-5 normal fibroblasts was negative. The proliferation of CNHK300 in breast cancer cells is similar to that of wild type adenovirus, and both of them have similar tumor cell killing ability. In MRC-5 normal fibroblasts, CNHK300 virus proliferative capacity was significantly weaker than that of wtAd5, at the same time, the destruction of normal fibroblasts was significantly weakened. CNHK300 virus and paclitaxel combined application, compared with the same dose of both alone, cell viability decreased significantly. The CNHK300 virus E1A can be selectively expressed in telomerase positive breast cancer cells and not in telomerase negative normal fibroblasts. CONCLUSION: CNHK300, a tumor-selective proliferative adenovirus, selectively replicates in some telomerase-positive breast cancer cells and produces oncolytic effects. Synergistic effect with paclitaxel chemotherapeutic agent.