子宫内膜癌患者NLRC5与自噬蛋白表达的相关性及其临床意义

Correlation of the expression of NLRC5 with levels of autophagy-associated proteins and its clinical significance in endometrial cancer

摘要:

目的 分析子宫内膜癌患者NLR家族含CARD结构域5（NLRC5）表达与自噬相关蛋白Beclin1、微管相关蛋白1轻链3（LC3）表达的相关性，并探讨NLRC5、Beclin1、LC3与子宫内膜癌预后的相关性。方法 采集2020年1月—2022年1月安徽医科大学第二附属医院90例子宫内膜癌患者癌组织作为子宫内膜癌组，另取90例正常子宫内膜作为子宫内膜组。采用实时荧光定量聚合酶链反应（qRT-PCR）测定两组NLRC5、Beclin1、LC3 mRNA的表达。子宫内膜癌组患者术后接受12个月的随访，按照实体瘤疗效标准分为预后良好组（72例）与预后不良组（18例）；比较两组NLRC5、Beclin1、LC3 mRNA的表达，采用多因素一般Logistic回归模型分析影响子宫内膜癌预后的因素，绘制受试者工作特征（ROC）曲线分析NLRC5、Beclin1、LC3对子宫内膜癌患者患者预后的预测价值。结果 子宫内膜癌组与正常子宫内膜组NLRC5、Beclin1和LC3 mRNA相对表达量的比较，差异均有统计学意义（P <0.05）；子宫内膜癌组NLRC5 mRNA相对表达量较正常子宫内膜组低，Beclin1和LC3 mRNA相对表达量较正常子宫内膜组高。NLRC5与Beclin1（r =-0.565，P <0.05）、LC3（r =-0.421，P <0.05）呈负相关；Beclin1与LC3呈正相关（r =0.462，P <0.05）。预后良好、预后不良组的年龄、体质量指数、脉管浸润、肿瘤直径比较，差异均无统计学意义（P >0.05）；预后不良组FIGO分期、宫体肌层深肌层占比、淋巴结转移、Beclin1、LC3 mRNA相对表达量高于预后良好组，NLRC5 mRNA相对表达量低于预后良好组（P <0.05）；FIGO分期Ⅱ、Ⅲ期［O^R =3.760（95% CI：1.260，11.223）］、宫体肌层深肌层［O^R =26.800（95% CI：7.043，101.984）］、有淋巴结转移［O^R =11.324（95% CI：3.286，39.019）］及Beclin1 ［O^R =68.163（95% CI：2.537，1831.270）］、LC3高表达［O^R =26.468（95% CI：1.444，485.127）］均是子宫内膜癌预后的危险因素（P <0.05）；NLRC5高表达［O^R =0.001（95% CI：0.000，0.079）］为保护因素（P <0.05）；ROC曲线分析结果显示，NLRC5敏感性为82.8%（95% CI：0.784，0.923）、特异性为73.9%（95% CI：0.670，0.938）；Beclin1敏感性为56.8%（95% CI：0.638，0.853）、特异性为77.8%（95% CI：0.644，0.877）；LC3敏感性为57.3（95% CI：0.680，0.815）、特异性为72.2%（95% CI：0.693，0.824）；联合敏感性为94.4%（95% CI：0.824，0.971）、特异性为62.5%（95% CI：0.593，0.778）。结论 子宫内膜癌患者NLRC5呈低水平表达，NLRC5与自噬相关蛋白Beclin1、LC3呈负相关，3者联合检测可提高子宫内膜癌患者的预后预测价值。

Abstract:

Objective To analyze the correlation between the expression of NLR family CARD domain containing 5 (NLRC5) and the levels of autophagy-associated proteins Beclin1 and microtubule-associated protein 1 light chain 3 (LC3) in patients with endometrial cancer, and to explore the correlations of NLRC5, Beclin1 and LC3 with the prognosis of endometrial cancer.Methods From January 2020 to January 2022, cancer tissues of 90 patients with endometrial cancer in the Second Affiliated Hospital of Anhui Medical University were collected and set as the endometrial cancer group, and normal endometrium was collected and set as the normal endometrial group. The expressions of NLRC5, Beclin1 and LC3 in the two groups were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Patients in the endometrial cancer group were followed up for 12 months after surgery, and were divided into good prognosis group (n = 72) and poor prognosis group (n = 18) according to the World Health Organization and Response Evaluation Criteria in Solid Tumors. The expressions of NLRC5, Beclin1 and LC3 in the good prognosis group and the poor prognosis group were compared. The multivariable Logistic regression model was established to analyze the factors affecting the prognosis of endometrial cancer, and the receiver operating characteristic (ROC) curve was plotted to analyze the predictive values of NLRC5, Beclin1 and LC3 for the prognosis of patients with endometrial cancer.Results There were significant differences in the expressions of NLRC5, Beclin1 and LC3 between the endometrial cancer group and the normal endometrial group (P < 0.05). Specifically, the expression of NLRC5 in the endometrial carcer group was lower than that in the normal endometrial group, while the expressions of Beclin1 and LC3 were higher in the endometrial carcer group than those in the normal endometrial group (P < 0.05). The expression of NLRC5 was negatively correlated with that of Beclin1 (r = -0.565, P < 0.05) and that of LC3 (r = -0.421, P < 0.05), and the expression of Beclin1 was positively correlated with that of LC3 (r = 0.462, P < 0.05). There was no significant difference in age, body mass index (BMI), vascular invasion and tumor diameter between patients in the good prognosis group and the poor prognosis group (P > 0.05). The FIGO stage, the proportion of deep muscular layer invasion in the uterine body, the frequency of lymph node metastasis, and the expressions of Beclin1 and LC3 in the poor prognosis group were higher than those in the good prognosis group, while the expression of NLRC5 in the poor prognosis group was lower than that in the good prognosis group (P < 0.05). FIGO stages II and III [O^R = 3.760 (95% CI: 1.260, 11.223) ], deep muscular layer invasion in the uterine body [O^R = 26.800 (95% CI: 7.043, 101.984) ], lymph node metastasis [O^R = 11.324 (95% CI: 3.286, 39.019) ], and high expressions of Beclin1 [O^R = 68.163 (95% CI: 2.537, 1831.270) ] and LC3 [O^R = 26.468 (95% CI: 1.444, 485.127)] were risk factors for the poor prognosis of endometrial cancer (P < 0.05), while the high expression of NLRC5 [O^R = 0.001 (95% CI: 0.000, 0.079) ] was a protective factor for the poor prognosis of endometrial cancer (P < 0.05). The ROC curve analysis showed that the sensitivity of NLRC5 for predicting the prognosis of endometrial cancer was 82.8% (95% CI: 0.784, 0.923), with the specificity being 73.9% (95% CI: 0.670, 0.938). The sensitivity and specificity of Beclin1 for predicting the prognosis of endometrial cancer were 56.8% (95% CI: 0.638, 0.853) and 77.8% (95% CI: 0.644, 0.877), while those of LC3 were 57.3% (95% CI: 0.680, 0.815) and 72.2% (95% CI: 0.693, 0.824). The sensitivity of the combined detection for predicting the prognosis of endometrial cancer was 94.4% (95% CI: 0.824, 0.971), with the specificity being 62.5% (95% CI: 0.593, 0.778).Conclusions NLRC5 is expressed at a low level in patients with endometrial cancer, and the expression of NLRC5 is inversely correlated with the levels of autophagy-associated proteins Beclin1 and LC3. The combination of the three yields a higher prognostic value for patients with endometrial cancer.