摘要
目的通过体外细胞实验,从分子水平探讨脑和肌肉芳香烃受体核转运样蛋白1(Bmal1)基因对结直肠癌细胞放疗敏感性的影响及其机制。方法体外培养结直肠癌细胞,运用RNA干扰技术转染Bmal1 siRNA,构建Bmal1基因敲除的结直肠癌细胞株。将体外培养的细胞分为对照组、Bmal1基因敲除组(敲除组)、放疗组、Bmal1基因敲除联合放疗组(联合组)。CCK-8检测各组结直肠癌细胞的增殖能力。RT-qPCR检测各组细胞Bmal1、缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的mRNA表达水平。Western blot检测各组细胞Bmal1、HIF-1α和VEGF的表达水平。结果CCK-8检测结果显示,细胞增殖能力由强到弱依次为:对照组、敲除组、放疗组、联合组,各组比较差异均有统计学意义(P<0.05)。RT-qPCR检测结果显示,联合组细胞HIF-1α和VEGF mRNA的表达水平在4组细胞中均为最低,分别为对照组的0.40倍和0.38倍,其次为放疗组、敲除组,对照组HIF-1α和VEGF mRNA表达水平最高,各组比较差异均有统计学意义(P<0.05)。Western blot检测结果显示,HIF-1α和VEGF在联合组细胞中表达水平最低,分别为对照组的0.45倍和0.35倍,其次为放疗组、敲除组,对照组HIF-1α和VEGF蛋白的表达水平最高,各组比较差异均有统计学意义(P<0.05)。结论敲除节律基因Bmal1可以增加结直肠癌细胞对放疗的敏感性,其机制可能与调节HIF-1α/VEGF信号通路有关。
Objective To investigate the effect of rhythm gene Bmal1 on the radiotherapy sensitivity of colorectal cancer cells and its mechanism at the molecular level through in vitro cell experiment.Methods Colorectal cancer cells were cultured in vitro and transfected with Bmal1 siRNA by RNA interference technique to construct colorectal cancer cell lines with Bmal1 gene knockout.The cells cultured in vitro were divided into the control group,Bmal1 gene knockout group(knockout group),radiotherapy group,Bmal1 gene knockout combined with radiotherapy group(combined group).CCK-8 was used to detect the proliferation ability of colorectal cancer cells in each group.The mRNA relative expression levels of Bmal1,HIF-1αand VEGF were detected by RT-qPCR.The expression levels of Bmal1,HIF-1αand VEGF were detected by Western Blot.Results The CCK-8 experiment results showed that the cell proliferation ability from strong to weak was in turn the control group,Bmal1 gene knockout group,radiotherapy group,combined group,and the differences were statistically significant(P<0.05).The RT-qPCR detection results showed that the expression levels of HIF-1αand VEGF mRNA in the combined group were the lowest in the four groups,which were 0.40 and 0.38 times of that in the control group,respectively,followed by the radiotherapy group and gene knockout group.The expression levels of HIF-1αand VEGF mRNA in the control group were the highest,and the difference was statistically significant(P<0.05).The Western Blot experiment results showed that the expression levels of HIF-1αand VEGF protein in the combined group were the lowest,which were 0.45 and 0.35 times of that in the control group,respectively,followed by the radiotherapy group and knockout group.The expression levels of HIF-1αand VEGF protein in the control group were the highest,and the difference was statistically significant(P<0.05).Conclusion Knockout of rhythm gene Bmal1 can increase the sensitivity of colorectal cancer cells to radiotherapy,and its mechanism may be related to th
作者
张博汝
魏柏
石晓雅
谢梦丽
ZHANG Boru;WEI Bai;SHI Xiaoya;XIE Mengli(Department of Oncology,Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430077,China)
出处
《重庆医学》
CAS
2022年第15期2544-2548,共5页
Chongqing medicine
基金
湖北省自然科学基金项目(20190101041)。
