促癌基因SNORA72对结直肠癌细胞放射敏感性的作用研究  

Oncogene SNORA72 enhances radioresistance in colorectal cancer cells

作　　者：张文成 邓佳荣 刘鑫 张宏 王治东 沈丽萍 Zhang Wencheng;Deng Jiarong;Liu Xin;Zhang Hong;Wang Zhidong;Shen Liping(Beijing Key Laboratory of Radiobiology,Institute of Radiation Medicine,Institute of Military Medical Research,Academy of Military Science,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院辐射医学研究所,北京市放射生物学重点实验室,北京100850

出　　处：《国际放射医学核医学杂志》2024年第2期99-113,共15页International Journal of Radiation Medicine and Nuclear Medicine

基　　金：国家自然科学基金面上项目(82373526)。

摘　　要：目的探索核仁小RNA(snoRNA)SNORA72基因在不同癌症特别是结直肠癌(CRC)中的表达模式及其对CRC细胞的生长及放射敏感性的影响。方法应用开放的癌症数据库分析SNORA72在不同癌症组织和CRC组织中的表达水平。构建过表达或敲低SNORA72的CRC细胞株HT29,将HT29细胞株分为过表达SNORA72组(LV-SNORA72)及其阴性对照组(LV-NC)、敲低SNORA72表达组(ASO-SNORA72)及其阴性对照组(ASO-NC)。采用实时荧光定量聚合酶链反应(qRT-PCR)检测HT29细胞中SNORA72表达情况。分别检测在体外过表达或敲低SNORA72后,对细胞增殖、细胞克隆形成、细胞凋亡、细胞周期的影响。对LV-SNORA72组及LV-NC组的HT29细胞进行不同剂量60Coγ射线照射,检测各组细胞的存活分数(SF)和凋亡率。采用转录组学分析法探讨SNORA72影响HT29细胞生长可能的作用机制。两组间的比较采用独立样本t检验。结果癌症数据库分析发现SNORA72在包括CRC在内的多种癌症组织中高表达,且差异均有统计学意义(均P<0.05)。qRT-PCR检测结果显示,与LV-NC组相比,LV-SNORA72组SNORA72的相对表达水平明显升高[(2.68±0.06)对(1.00±0.17)],且差异有统计学意义(t=16.570,P<0.001)。另外,与ASO-NC组相比,ASO-SNORA72组SNORA72的相对表达水平明显降低[(0.61±0.08)对(1.00±0.13)],且差异有统计学意义(t=4.355,P<0.05)。细胞增殖检测结果显示,在实验的第3、4、5天,LV-SNORA72组的吸光度值明显高于LV-NC组[(0.79±0.05)对(0.51±0.09)、(1.78±0.04)对(1.22±0.05)、(3.30±0.05)对(2.19±0.06)],且差异均有统计学意义(t=8.582、16.400、31.200,均P<0.001)。相反,ASO-SNORA72组的吸光度值明显低于ASO-NC组[(0.42±0.07)对(0.55±0.05)、(1.04±0.08)对(1.25±0.05)、(1.46±0.09)对(1.74±0.08)],且差异均有统计学意义(t=3.957、6.147、8.471,均P<0.01)。细胞克隆形成实验结果显示,LV-SNORA72组的克隆形成率明显高于LV-NC组[(40.87±1.70)%对(26.60±0.40)%],且差异有统计学意义(tObjective To explore the expression patterns of small nucleolar RNA(snoRNA)SNORA72 gene in various cancers,particularly in colorectal cancer(CRC),and its effect on the growth and radiosensitivity of CRC cells.Methods The expression of SNORA72 in different cancer and CRC tissues was analyzed using open cancer databases.The CRC cell line HT29,overexpressing or knocking down SNORA72,was constructed,dividing HT29 cells into the overexpressing SNORA72 group(LV-SNORA72)and its negative control group(LV-NC),as well as the SNORA72 knockdown group(ASO-SNORA72)and its negative control group(ASO-NC).The expression of SNORA72 in HT29 cells was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The effects of SNORA72 overexpression or knockdown on cell proliferation,colony formation,apoptosis,and cell cycle were evaluated.The HT29 cells from the LV-SNORA72 and LV-NC groups were irradiated with different doses of 60Coγ-rays,and the survival fraction(SF)and apoptosis rate of cells in each group were assessed.Transcriptomic analysis was employed to explore the potential mechanisms by which SNORA72 affects HT29 cell growth.An independent sample t-test was used for comparisons between two groups.Results Analysis of cancer databases revealed that SNORA72 is overexpressed in various cancer tissues,including CRC,the difference were statistically significant(all P<0.05).qRT-PCR results indicated that the relative expression of SNORA72 in the LV-SNORA72 group was significantly higher than that in the LV-NC group((2.68±0.06)vs.(1.00±0.17)),and the difference was statistically significant(t=16.570,P<0.001).Conversely,the relative expression of SNORA72 in the ASO-SNORA72 group was significantly lower than that in the ASO-NC group((0.61±0.08)vs.(1.00±0.13)),and the difference was statistically significant(t=4.355,P<0.05).Cell proliferation assay results showed that the absorbance values of the LV-SNORA72 group were significantly higher than those of the LV-NC group((0.79±0.05)vs.(0.51±0.09),(1.78±0.0

关 键 词：RNA 小核仁 结直肠肿瘤 辐射耐受性 基因表达调控 SNORA72基因 

分 类 号：R735.34[医药卫生—肿瘤]