宫颈癌组织中NEK2 和circPITX1 的表达水平与放疗敏感度及预后的相关性研究

目的 探讨宫颈癌组织中中心体相关激酶2(never in mitosis geneA related kinase-2，NEK2) 和环状RNA 配对同源结构域基因1(circRNA paired homology domain gene 1，circPITX1) 表达与放疗敏感度及预后的关系。方法 选取2017 年1月～ 2018 年1 月四川大学华西三亚医院接受放疗治疗的82 例宫颈癌患者为研究对象。依据放疗后病灶退缩情况评价放疗疗效,分为放疗抵抗组（n=51）和放疗敏感组（n=31）。应用免疫组织化学法（immunohistochemistry，IHCS）检测宫颈癌癌组织和癌旁组织中NEK2 蛋白表达。采用荧光实时定量PCR（real-time quantitative PCR，RT-qPCR）检测circPITX1mRNA 和NEK2mRNA 表达。统计学分析癌组织中circPITX1mRNA 和NEK2 mRNA 与放疗敏感度、临床病理特征的关系。采用 Kaplan-Meier 生存曲线分析宫颈癌患者circPITX1mRNA 和NEK2 mRNA 表达水平与患者生存预后的关系。COX 回归分析宫颈癌患者预后不良的危险因素。结果 宫颈癌癌组织中NEK2 蛋白表达阳性率85.37%（70/82）] 高于癌旁组织12.20%（10/82）]，差异有统计学意义（χ2=87.857，P ＜ 0.05）。癌组织中circPITX1mRNA（1.92±0.24），NEK2 mRNA 表达（2.04±0.31）高于癌旁组织（0.81±0.20，0.93±0.27）表达，差异均有统计学意义（t=32.174，24.450，均P ＜ 0.05）。宫颈癌组织中NEK2 mRNA 与circPITX1mRNA 表达呈显著正相关(r=0.618, P ＜ 0.05)。FIGO 分期Ⅲ A～Ⅲ B 期宫颈癌患者NEK2 mRNA（2.54±0.23），circPITX1mRNA（2.33±0.19）表达明显高于IIB 期患者（1.17±0.40，1.21±0.30）表达，差异均有统计学意义（t=19.730, 20.710,均P ＜ 0.05）。宫颈癌放疗抵抗组NEK2 mRNA（2.39±0.38），circPITX1mRNA（2.50±0.21）均显著高于放疗敏感组（1.83±0.28，0.97±0.28），差异有统计学意义（t=7.109, 28.149, 均P ＜ 0.05）。NEK2 mRNA（48.78%（20/41）] 和circPITX1mRNA54.76%（23/42）] 高表达组的三年总生存率显著低于NEK2 mRNA90.24%（37/41）]，circPITX1 mRNA 低表达组 85.00%（34/40）]，差异均有统计学意义（χ2=5.118, 4.890, 均P ＜ 0.05）；NEK2 mRNA（26.10±3.62）个月] 和circPITX1mRNA（25.87±3.81）个月]高表达组的平均生存时间显著短于NEK2 mRNA（ 33.25±4.09）个月]和circPITX1mRNA低表达组（ 32.02±4.02）个月]，差异均有统计学意义（t=8.382, 7.140, 均P ＜ 0.05）。FIGO 分期Ⅲ A～Ⅲ B（HR=3.754，95%CI:1.621～ 8.694），NEK2 mRNA高表达（HR=2.874，95%CI:2.504 ～ 3.497），circPITX1mRNA 高表达（HR=2.679，95%CI:2.485 ～ 3.184）是宫颈癌患者预后不良的独立危险因素（均P 均＜ 0.05）。结论 NEK2 和circPITX1 在宫颈癌组织中表达升高, 可成为放疗敏感度及预后预测的标志物。

Correlation between the Expression Levels of NEK2 and CircPITX1 and Radiotherapy Sensitivity and Prognosis in Cervical Cancer

Objective To explore the expression never in mitosis gene A-related kinase 2 (NEK2) and circRNA paired homology domain gene 1 (circPITX1) in cervical cancer tissues and their correlation with radiotherapy sensitivity and prognosis. Methods 82 patients with cervical cancer who received radiotherapy in West China Sanya Hospital of Sichuan University from January 2017 to January 2018 were selected as research object. The efficacy of radiotherapy was evaluated according to the lesion shrinkage after radiotherapy, and the patients were divided into radiotherapy resistant group (n=51) and radiotherapy sensitive group (n=31). The expression of NEK2 protein in cervical cancer tissues and adjacent tissues was detected by immunohistochemistry（iIHCS）. The expression of circPITX1mRNA and NEK2 mRNA was detected by fluorescence realtime quantitative PCR（RT-qPCR）. The relationship between circPITX1 mRNA and NEK2 mRNA in cancer tissue with radiosensitivity and clinicopathological characteristics was statistically analyzed. The Kaplan-Meier survival curve was used to analyze the relationship between the expression levels of circPITX1mRNA and NEK2 mRNA and the 3-year prognosis of cervical cancer patients.COX regression analysis was used to analyze the risk factors of poor prognosis in patients with cervical cancer. Results The positive rate of NEK2 protein expression in cervical cancer tissues 85.37% (70/82)] was higher than that in adjacent tissues 12.20% (10/82)], and the difference was statistically significant (χ2=87.857, P ＜ 0.05). The expressions of circPITX1mRNA (1.92±0.24) and NEK2 mRNA (2.04±0.31) in cancer tissues were higher than those of circPITX1 mRNA (0.81±0.20) and NEK2 mRNA (0.93±0.27) paracancerous tissues, and the differences were statistically significant (t=32.174, 24.450, all P ＜ 0.05). There was a significant positive correlation between NEK2 mRNA and CircPITX1mRNA expression in cervical cancer tissue (r=0.618, P ＜ 0.05). The expressions of NEK2 mRNA (2.54±0.23) and circPITX1mRNA (2.33±0.19) in patients with FIGO stage IIIA ～ IIIB cervical cancer were significantly higher than those in IIB stage patients (1.17±0.40) and CircPITX1 mRNA(1.21±0.30)，and the differences were statistical significance (t=19.730, 20.710, all P ＜ 0.05). The NEK2 mRNA (2.39±0.38) and circPITX1mRNA (2.50±0.21) in the radiotherapy-resistant cervical cancer group were significantly higher than those in the radiosensitive group (1.83±0.28, 0.97±0.28), and the differences were statistically significant (t =7.109, 28.149，all P ＜ 0.05). The 3-year overall survival rate of NEK2 mRNA (48.78% (20/41)], circPITX1mRNA 54.76% (23/42)] high expression group was significantly lower than that of NEK2 mRNA 90.24% (37/41)], and CircPITX1mRNA low expression group 85.00% (34/40)], and the differences were statistically significant (χ2=5.118, 4.890, all P ＜ 0.05). The mean survival time of NEK2 mRNA (26.10±3.62) months] and circPITX1mRNA (25.87±3.81) months] high expression group was significantly shorter than that of NEK2 mRNA (33.25±4.09) months] and the circPITX1mRNA low expression group (32.02±4.02) months], and the differences were statistically significant (t=8.382, 7.140，all P ＜ 0.05). FIGO stage IIIA ～ IIIB (HR=3.754, 95%CI: 1.621 ～ 8.694), high expression of NEK2 mRNA (HR=2.874, 95%CI: 2.504 ～ 3.497), high expression of circPITX1mRNA (HR=2.679, 95%CI: 2.485 ～ 3.184) were independent risk factors for poor prognosis of cervical cancer patients (all P ＜ 0.05). Conclusion NEK2 and circPITX1were up-regulated in cervical cancer tissues and could be used as markers for radiosensitivity and prognosis prediction.