| 倪猛,殷涛,王旸,等.抑制FOXD1基因表达增强结直肠癌细胞HCT116的放射敏感性[J].中华放射医学与防护杂志,2018,38(12):886-893.Ni Meng,Yin Tao,Wang Yang,et al.Inhibition of FOXD1 gene expression increases radiosensitivity of colorectal cancer HCT116 cells[J].Chin J Radiol Med Prot,2018,38(12):886-893 |
| 抑制FOXD1基因表达增强结直肠癌细胞HCT116的放射敏感性 |
| Inhibition of FOXD1 gene expression increases radiosensitivity of colorectal cancer HCT116 cells |
| 投稿时间：2018-06-29 |
| DOI：10.3760/cma.j.issn.0254-5098.2018.12.002 |
| 中文关键词: FOXD1基因 结直肠癌 放射敏感性 |
| 英文关键词:FOXD1 gene Colorectal cancer Radiosensitiviy |
| 基金项目:国家自然科学基金（81372665） |
|
| 摘要点击次数: 3419 |
| 全文下载次数: 1728 |
| 中文摘要: |
| 目的 研究抑制FOXD1基因的表达对结直肠癌细胞放射敏感性的影响。方法 采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blot检测人结直肠癌组织和细胞中FOXD1 mRNA和蛋白的表达。对结直肠癌HCT116细胞行梯度剂量（0、2、4、6 Gy）X射线照射，qRT-PCR和Western blot检测各组细胞中FOXD1的表达。将siRNA阴性对照和FOXD1 siRNA转染至结直肠癌细胞中，分别记为si-NC组和si-FOXD1组，经4 Gy的X射线照射处理后记为si-NC+4 Gy组和si-FOXD1+4 Gy组，Western blot检测各组细胞中FOXD1的表达，四甲基偶氮唑盐（MTT）法检测各组细胞的增殖活性，克隆形成实验检测各组细胞存活率，采用TECT DNA-PK试剂盒检测各组细胞中DNA-PK活性，将转染的结直肠癌细胞接种于BALB/c裸鼠建立移植瘤模型，进行射线照射后，检测各组肿瘤的体积和质量变化。结果 与癌旁正常组织相比，结直肠癌组织中FOXD1 mRNA和蛋白的表达均显著增加（t=5.579、4.816，P<0.05），与结肠黏膜上皮细胞NCM460相比，结直肠癌细胞株中FOXD1 mRNA（t=5.85~17.62，P<0.05）和蛋白（t=9.04~11.42，P<0.05）表达均显著升高。结直肠癌细胞HCT116中FOXD1的表达量随着放射剂量增加而升高，呈现剂量依赖性，差异有统计学意义（t=9.13~44.15，P<0.05）。转染si-FOXD1能够有效地抑制结直肠癌细胞中FOXD1的表达（t=10.51，P<0.05），FOXD1敲低后能够抑制结直肠癌细胞的增殖活性（t=10.41，P<0.05），提高结直肠癌细胞的放射敏感性，放射增敏比为1.797，降低放射诱导的DNA-PK的活性（t=6.20，P<0.05）。抑制FOXD1的表达经射线照射后，裸鼠种植瘤体积和重量明显减小（t=11.29、3.69，P<0.05）。结论 抑制FOXD1基因的表达能够提高结直肠癌细胞的放射敏感性，抑制结直肠癌裸鼠移植瘤的生长，可为改善放射治疗对结直肠癌患者的治疗效果提供潜在的靶向基因。 |
| 英文摘要: |
| Objective To study the effect of inhibiting the expression of FOXD1 gene on the radiosensitivity of colorectal cancer cells.Methods The expressions of FOXD1 mRNA and protein in human colorectal cancer tissues and cells were detected by Real-time PCR (qRT-PCR) and Western blot. The colorectal cancer cell line HCT116 was irradiated with 0, 2, 4 and 6 Gy of X-rays. The expression of FOXD1 in each groups were detected by qRT-PCR and Western blot. HCT116 cells were transfected with FOXD1 siRNA and its negative control and termed as si-FOXD1 group and si-NC group. When these cells were irradiated with 4 Gy X-rays, they were termed as si-FOXD1+4 Gy group and si-NC+4 Gy group. Cell proliferation was detected with MTT method, cell survival fraction was measured with colony formation assay, and DNA-PK activity was detected by TECT DNA-PK kit. The siRNA-transfected colorectal cancer cells were inoculated into BALB/c nude mice to establish the xenograft model. After irradiation, the volume and quality of the subcutaneous transplanted tumors were measured every 5 days.Results The expression of FOXD1 mRNA and protein in colorectal cancer tissues was higher than that in adjacent normal tissues (t=5.579, 4.816, P<0.05). The mRNA(t=5.85-17.62, P<0.05) and protein(t=9.04-11.42, P<0.05) expression of FOXD1 in different colorectal cancer cell lines was higher than that in colonic mucosa epithelial cell line NCM460. The expression of FOXD1 in colorectal cancer cells HCT116 was increased after radiation in a dose dependent manner(t=9.13-44.15, P<0.05). Transfection of si-FOXD1 effectively inhibited the expression of FOXD1 (t=10.51, P<0.05), decreased proliferation (t=10.41, P<0.05), increased radiosensitivity with a radiosensitization ratio of 1.797, and reduced the radiation-induced DNA-PK activity (t=6.20, P<0.05) in colorectal cancer cells. After localized irradiation, the tumor volume and weight in nude mice transplanted with si-FOXD1 HCT116 cells were significantly smaller than those in HCT116 (t=11.29, 3.69, P<0.05).Conclusions Knock-down of FOXD1 gene increases the radiosensitivity of colorectal cancer cells and inhibits the growth of colorectal cancer xenograft in nude mice, which provides a potential target gene in improving the effect of radiotherapy on colorectal cancer. |
| HTML 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|