目的:研究结直肠癌(Colorectal cancer, CRC)患者血浆和癌组织中microRNA-181和张力蛋白同源物基因(PTEN)的表达及其与临床分期,并探讨CRC在结直肠癌中的诊断价值。方法:选择120例结直肠癌患者及100例健康体检者为对照。采用实时荧光定量PCR技术检测结直肠癌患者血浆中miRNA-181和PTEN基因、以及组织中miRNA-181的表达,采用免疫组织化学技术检测结直肠癌患者血浆和组织PTEN蛋白的表达,并分析miRNA-181和PTEN蛋白表达与结直肠癌病理分期的关系。采用四甲基偶氮唑盐法和细胞集落形成实验研究miRNA-181和PTEN在CRC细胞株SW 1116生长和增殖能力。结果:与正常对照组比较,结直肠癌患者血浆中miRNA-181明显升高(0.58 ± 0.23 vs 3.28 ± 0.46 (P < 0.01),血浆中PTEN基因明显下降(5.37 ± 1.34 vs 0.26 ± 0.21) (P = 0.01)。与癌旁正常结直肠粘膜组织比较,结直肠癌组织PTEN蛋白阳性表达率明显降低(100% vs 36.67%) (P < 0.001);随着分期恶性度的升高,miRNA-181相对表达量增加,PTEN基因和蛋白的表达下降,各组间比较差异均有统计学意义(P < 0.01, P < 0.001)。四甲基偶氮唑盐法显示miRNA-181组在不同时间的CRC细胞株SW 1116生长速度加快,PTEN组细胞生长落后(P < 0.01, P < 0.001, P < 0.001)。平板克隆形成实验显示miRNA-181组在CRC细胞株SW 1116增殖能力增强,PTEN组细胞增殖能力减弱(P < 0.001)。结论:miRNA-181和PTEN在CRC血浆和组织中的异常表达与结直肠癌病理分期密切相关,miRNA-181对结直肠癌的辅助诊断和预后监测具有重要的临床意义。
Objective: To investigate the relationship between plasma microRNA (miRNA)-181 and tension protein homolog gene (PTEN) and pathological staging of colorectal cancer, and to explore the di-agnosis significance for colorectal cancer. Methods: A total of 120 cases of colorectal cancer and 100 cases of healthy subjects were included in the study. Real-time fluorescence quantitative PCR was used to detect the expression of miRNA-181 and PTEN genes in the plasma and tissues of colorectal cancer patients, and immunohistochemistry was used to detect the expression of PTEN protein in the tissues of colorectal cancer patients, and to analyze the relationship between the expression of miRNA-181and PTEN protein and the pathological stage of colorectal cancer. The growth and proliferation capacity of miRNA-181 and PTEN in CRC cell line SW 1116 were studied by 4-Methylcyclohexanone assay (MTT) and cell colony formation assay. Results: Compared with the normal control group, the plasma level of miRNA-181 in colorectal cancer patients was significantly increased (0.58 ± 0.23 vs 3.28 ± 0.46) vs (P < 0.01), and the plasma level of PTEN gene in colorectal cancer patients was significantly decreased (5.37 ± 1.34 vs 0.26 ± 0.21) (P < 0.01). The positive expression rate of PTEN protein in colorectal cancer tissues was significantly decreased (100% vs 36.67%) (P < 0.001). With the increase of malignant degree of staging, the relative expression level of miRNA-181 increased, and the expression level of PTEN gene and protein decreased, with statistically significant differences between groups (P < 0.01, P < 0.001). The MTT showed that CRC cell line SW1116 in the miRNA-181 group at different times increased the Growth rate, while the cell growth in the PTEN group lagged behind (P < 0.01, P < 0.001, P < 0.001). The cell colony formation experiment showed that the proliferation ability of CRC cell line SW1116 in the miRNA-181 group was enhanced, while the proliferation capacity of cells in the PTEN group was weakened (P < 0.001). Conclusion: The abnormal expression of miRNA-181 and PTEN in CRC plasma and tissues is closely related to the pathological stage of colorectal cancer, And miRNA-181 has important clinical significance in the auxiliary diagnosis and prognosis monitoring of colorectal cancer.
结直肠癌,miRNA-181,张力蛋白同源物基因(PTEN), Colorectal Cancer microRNA-181 Tension Protein Homolog Gene (PTEN)结直肠癌血浆中microRNA-181和PTEN基因与病理分期的关系
郭伟强,何凤屏*,张涛,李定云,黄亮,马占忠,陈红,刘玉兰,陈伟娟,陈卓然
广东省汕头大学医学院附属粤北人民医院,广东 汕头
收稿日期:2020年5月11日;录用日期:2020年5月27日;发布日期:2020年6月2日
摘 要
目的:研究结直肠癌(Colorectal cancer, CRC)患者血浆和癌组织中microRNA-181和张力蛋白同源物基因(PTEN)的表达及其与临床分期,并探讨CRC在结直肠癌中的诊断价值。方法:选择120例结直肠癌患者及100例健康体检者为对照。采用实时荧光定量PCR技术检测结直肠癌患者血浆中miRNA-181和PTEN基因、以及组织中miRNA-181的表达,采用免疫组织化学技术检测结直肠癌患者血浆和组织PTEN蛋白的表达,并分析miRNA-181和PTEN蛋白表达与结直肠癌病理分期的关系。采用四甲基偶氮唑盐法和细胞集落形成实验研究miRNA-181和PTEN在CRC细胞株SW 1116生长和增殖能力。结果:与正常对照组比较,结直肠癌患者血浆中miRNA-181明显升高(0.58 ± 0.23 vs 3.28 ± 0.46 (P < 0.01),血浆中PTEN基因明显下降(5.37 ± 1.34 vs 0.26 ± 0.21) (P = 0.01)。与癌旁正常结直肠粘膜组织比较,结直肠癌组织PTEN蛋白阳性表达率明显降低(100% vs 36.67%) (P < 0.001);随着分期恶性度的升高,miRNA-181相对表达量增加,PTEN基因和蛋白的表达下降,各组间比较差异均有统计学意义(P < 0.01, P < 0.001)。四甲基偶氮唑盐法显示miRNA-181组在不同时间的CRC细胞株SW 1116生长速度加快,PTEN组细胞生长落后(P < 0.01, P < 0.001, P < 0.001)。平板克隆形成实验显示miRNA-181组在CRC细胞株SW 1116增殖能力增强,PTEN组细胞增殖能力减弱(P < 0.001)。结论:miRNA-181和PTEN在CRC血浆和组织中的异常表达与结直肠癌病理分期密切相关,miRNA-181对结直肠癌的辅助诊断和预后监测具有重要的临床意义。
选取2017年1月至2019年6月在粤北人民医院胃肠外科住院并病理诊断证实为结直肠癌的患者120例,其中男68例,女52 例,年龄47~70岁,平均年龄55.6岁。所有的患者术前均未接受放射治疗、化学药物、免疫治疗。纳入标准:① 病理确诊为结直肠癌的患者;② 未经任何放疗、化疗等治疗的患者。排除标准:① 合并有其他恶性肿瘤病史;② 家族遗传性息肉病患者。根据美国癌症联合委员会(American Joint Committee on Cancer, AJCC)/国际抗癌联盟(the International Union Against Cancer, UICC)的结直肠癌TNM分期标准 [11]:I期10例,II期18例,III期48例,IV期44例;另取同期健康体检者100例作为对照,年龄45岁~65岁,平均年龄52.28岁,男50例,女50例。两组人群年龄差异无统计学意义。本研究经汕头大学医学院附属粤北人民医院伦理委员会批准,所有研究对象均签约知情同意书。
郭伟强,何凤屏,张 涛,李定云,黄 亮,马占忠,陈 红,刘玉兰,陈伟娟,陈卓然. 结直肠癌血浆中microRNA-181和PTEN基因与病理分期的关系Relationship between Plasma miRNA-181 and PTEN and Pathological Staging in Colorectal Cancer[J]. 医学诊断, 2020, 10(02): 56-65. https://doi.org/10.12677/MD.2020.102009
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