Department of Oncology, Tangdu Hospital, Fourth Military Medical University, 1 Xinsi Road, Xi'an 710038, China.
J Cancer Res Clin Oncol. 2012 Aug;138(8):1329-38. doi: 10.1007/s00432-012-1206-2. Epub 2012 Apr 6.
To explore the relationship between the expression of ZEB1 gene and the proliferation ability of lung adenocarcinoma cells.
Immunohistochemistry, Western blot and Real-time PCR were used to detect the expression of ZEB1 gene in lung adenocarcinoma tissue and cell lines compared with adjacent noncancerous region and the human lung fibroblast cell HLF cells. The lentivirus RNA interference technique was used to knock down the expression of ZEB1 in lung adenocarcinoma A549 and H1299 cell lines. Cell cycle and cell apoptosis were measured by FCM assay. In vivo, four groups of 4-week-old nude mice were subcutaneously injected with the stably transfected (ZEB-si, scr-si) cells at a single site to investigate the effect of ZEB1-siRNA in the nude mice tumor growth. In situ apoptosis was detection by TUNEL assay.
ZEB-1 was highly expressed in lung adenocarcinoma tissue and cell lines compared with adjacent noncancerous region and the human lung fibroblast cell HLF cells. ZEB1-siRNA could decrease lung adenocarcinoma cell proliferation by delaying S-phase entry and induce cell apoptosis, which led to the inhibition of the tumorigenicity of A549 and H1299 cell lines. Further investigation showed that injecting the ZEB1-siRNA cells into the nude mice could significantly decrease the tumor growth.
Knockdown of ZEB-1 expression by lentivirus-delivered siRNA may provide a novel therapeutic target for the treatment of lung cancer.
探讨 ZEB1 基因表达与肺腺癌细胞增殖能力的关系。
采用免疫组化、Western blot 和实时 PCR 检测 ZEB1 基因在肺腺癌组织及细胞系中与相邻非癌区及人肺成纤维细胞 HLf 细胞的表达差异。采用慢病毒 RNA 干扰技术敲低肺腺癌 A549 和 H1299 细胞系中 ZEB1 的表达。FCM 检测细胞周期和细胞凋亡。体内实验中，将稳定转染（ZEB-si、scr-si）细胞的 4 周龄裸鼠每组 4 只，在单个部位皮下注射，观察 ZEB1-siRNA 对裸鼠肿瘤生长的影响，采用 TUNEL 法检测原位细胞凋亡。
ZEB-1 在肺腺癌组织及细胞系中高表达，明显高于相邻非癌区及人肺成纤维细胞 HLf 细胞。ZEB1-siRNA 可通过延迟 S 期进入来减少肺腺癌细胞增殖并诱导细胞凋亡，从而抑制 A549 和 H1299 细胞系的致瘤性。进一步研究表明，向裸鼠注射 ZEB1-siRNA 细胞可显著抑制肿瘤生长。
慢病毒介导的 siRNA 敲低 ZEB-1 表达可能为肺癌的治疗提供新的治疗靶点。
