| 孙翊乐,祁钰雯,顾琳萍,牛晓敏,陈智伟.HOXD9促进肺癌发生发展的生物学作用及初步机制[J].现代生物医学进展英文版,2022,(13):2401-2410. |
| HOXD9促进肺癌发生发展的生物学作用及初步机制 |
| Biological Effect and Preliminary Mechanism of HOXD9 on the Development of Lung Cancer |
| Received:January 03, 2022 Revised:January 27, 2022 |
| DOI：10.13241/j.cnki.pmb.2022.13.001 |
| 中文关键词: 同源盒基因D9 肺癌 DNA损伤 增殖细胞核抗原 转录 |
| 英文关键词: HOXD9 Lung cancer DNA damage PCNA Transcription |
| 基金项目:国家自然科学基金面上项目（81572250、81972187）；上海申康医院发展中心项目（16CR3071B）；上海市科学技术委员会项目（19ZR1449800；20Y11913700） |
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| 中文摘要: |
| 摘要 目的：探究同源盒基因HOXD9在H3255、H2170肺癌细胞株中的生物学作用及作用机制。方法：构建HOXD9过表达稳转H3255、H2170肺癌细胞株，通过CCK-8、克隆形成、小鼠皮下瘤实验检测HOXD9对肺癌细胞增殖的影响；利用transwell、基质胶实验检测HOXD9对肺癌细胞迁移及侵袭的影响；采用彗星实验、磷酸化组蛋白H2AX（γH2AX）免疫荧光染色检测HOXD9对肺癌细胞DNA损伤的影响；通过流式细胞术检测HOXD9对肺癌细胞凋亡的影响；通过qRT-PCR、western blot、luciferase、ChIP、rescue实验探究HOXD9在肺癌中促癌的作用机制。结果：CCK-8、克隆形成、小鼠皮下瘤实验显示HOXD9可促进肺癌细胞增殖；transwell实验表明HOXD9可促进肺癌细胞迁移；基质胶实验显示HOXD9可促进肺癌细胞侵袭；彗星实验与γH2AX免疫荧光染色提示HOXD9可抑制肺癌细胞DNA损伤；流式细胞术显示HOXD9可抑制肺癌细胞凋亡。HOXD9可通过转录上调肺癌细胞中增殖细胞核抗原PCNA的表达水平。结论：HOXD9可通过促进PCNA的转录介导肺癌细胞的增殖、迁移、侵袭，抑制肺癌细胞DNA损伤及凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the biological effect and mechanism of homeobox D9 (HOXD9) in H3255 and H2170 lung cancer cell lines. Methods: H3255 and H2170 lung cancer cell lines with HOXD9 stable overexpression were constructed. Effect of HOXD9 on lung cancer cell proliferation was detected by CCK-8, colony formation assay and tumor xenograft experiments. Effect of HOXD9 on lung cancer cell migration and invasion was detected by transwell and matrigel assay. Effect of HOXD9 on lung cancer cell DNA damage was detected by comet assay and γH2AX immunofluorescence staining. Effect of HOXD9 on lung cancer cell apoptosis was detected by flow cytometry. The mechanism of HOXD9 in lung cancer was investigated by qRT-PCR, western blot, luciferase, ChIP and rescue assay. Results: CCK-8, colony formation assay and tumor xenograft experiments showed that HOXD9 promoted the proliferation of lung cancer cells. Transwell assay showed that HOXD9 promoted the migration of lung cancer cells. Matrigel assay showed that HOXD9 promoted the invasion of lung cancer cells. Comet assay and γH2AX immunofluorescence staining showed HOXD9 inhibited DNA damage of lung cancer cells. Flow cytometry showed HOXD9 inhibited the apoptosis of lung cancer cells. HOXD9 up-regulated proliferating cell nuclear antigen(PCNA) expression level in lung cancer cells through transcription regulation. Conclusion: HOXD9 promoted the proliferation, migration and invasion while inhibited DNA damage and apoptosis of lung cancer cells by inducing PCNA transcription. |
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