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目的:观察选择性增殖腺病毒CNHK500对乳腺癌的特异性杀伤作用。方法:行病毒增殖实验和细胞生长抑制实验,验证CNHK500选择性复制和杀伤能力;Westernblot检测腺病毒E1A和E1B在细胞中的表达。结果:CNHK500在乳腺癌细胞中复制能力与野生型腺病毒wtAd5相似,较ONYX-015增殖能力强。在正常成纤维细胞中CNHK500病毒增殖能力明显减弱,较wtAd5增殖能力弱1000倍左右。CNHK500可有效杀伤乳腺癌细胞株;而CNHK500对正常成纤维细胞的杀伤力较wtAd5减弱约100倍。CNHK500病毒的E1A可以选择性在端粒酶阳性的乳腺癌细胞株中表达,在端粒酶阴性的正常成纤维细胞株BJ中不表达,CNHK500可以选择性地在缺氧条件下表达E1B。动物实验结果显示,静脉注射CNHK500可以显著抑制MCF-7乳腺癌细胞裸鼠移植瘤的生长,治疗效果与给药剂量相关。结论:肿瘤选择性增殖腺病毒CNHK500可选择性在端粒酶阳性的乳腺癌细胞中复制,并产生体内外杀伤作用。
Objective: To observe the specific killing effect of selective adenovirus CNHK500 on breast cancer. Methods: The virus proliferation test and cell growth inhibition test were performed to verify the selective replication and cytotoxicity of CNHK500. Western blot was used to detect the expression of E1A and E1B in the cells. Results: The replication ability of CNHK500 in breast cancer cells was similar to wtAd5 of wild-type adenovirus and stronger than that of ONYX-015. In normal fibroblasts CNHK500 proliferation significantly weakened virus proliferation than wtAd5 weak about 1000 times. CNHK500 can effectively kill breast cancer cell lines; and CNHK500 on normal fibroblasts lethal than wtAd5 attenuated about 100 times. E1A of CNHK500 virus can be selectively expressed in telomerase positive breast cancer cell lines, but not in telomerase negative normal fibroblast strain BJ. CNHK500 can selectively express E1B under hypoxic conditions. Animal experiments show that intravenous injection of CNHK500 can significantly inhibit the growth of MCF-7 breast cancer xenografts in nude mice, and the therapeutic effect is related to the dosage. Conclusion: The tumor-selective adenovirus CNHK500 selectively replicates in telomerase-positive breast cancer cells and produces in vitro and in vivo cytotoxicity.