目的

探讨双肾上腺皮质激素样激酶（DCLK）1在子宫内膜样腺癌（EA）患者中的表达，并评估其对EA 患者临床特征的潜在影响。

方法

选择2019年1月至6月在四川大学华西第二医院妇科就诊的95例EA 患者，25例子宫内膜非典型增生（AH）患者及30例因子宫良性疾病进行子宫切除术的子宫内膜组织正常患者为研究对象，分别纳入EA 组（n=95）、AH 组（n=25）及对照组（n=30）。采用免疫组化方法，检测EA 组患者EA 组织、AH 组患者AH 组织和对照组正常子宫内膜组织中DCLK1蛋白表达水平。在EA 组EA 细胞系HEC-1-A 和Ishikawa中，转染过表达DCLK1质粒、空载体质粒，分别纳入过表达DCLK1 亚组、空载体亚组，将未转染质粒的HEC-1-A 和Ishikawa细胞系，纳入空白对照亚组。检测上述3 个亚组的HEC-1-A 和Ishikawa细胞DCLK1 与AKT/m TOR 信号通路中关键蛋白表达水平。本研究遵循的程序符合四川大学华西第二医院伦理委员会所制定的伦理学标准（批准文号：2022-038号），3组患者对本研究均知情同意，并均签署临床研究知情同意书。

结果

①EA 组、AH 组、对照组患者异常子宫出血占比比较，差异有统计学意义（χ²=45.90，P＜0.001）；这3 组患者子宫内膜腺体DCLK1 阳性表达率分别为83.2%（79/95）、76.0%（19/25）、40.0%（12/30），并且差异亦有统计学意义（χ²=21.83，P＜0.001）。②EA 组患者中，EA 组织不同病理学分级患者的DCLK1阳性表达率比较，差异有统计学意义（χ²=14.70，P=0.003），并且DCLK1表达水平与EA 组织病理学分级呈正相关关系（r_s=0.503，P＜0.001）。③过表达DCLK1亚组HEC-1-A、Ishikawa细胞中，DCLK1蛋白表达水平均较空载体亚组与空白对照亚组显著增高，蛋白激酶B（AKT）、磷酸化蛋白激酶B（p-AKT）和哺乳动物雷帕霉素靶蛋白（m TOR）蛋白表达水平，均较空载体亚组显著增高，并且差异均有统计学意义（P＜0.05）。

结论

DCLK1表达水平与EA 恶性表型密切相关，并且可能与AKT/m TOR 信号通路相关。DCLK1可能作为EA 患者临床诊断的潜在生物标志物和治疗靶点。

Objective

To investigate the expression of doublecortin-like kinase （DCLK）1 in endometrioid adenocarcinoma（EA）patients and evaluate its influence on the clinical characteristics of EA patients.

Methods

A total of 95 EA patients，25 patients with atypical hyperplasia（AH），and 30 with normal endometrial tissue who underwent hysterectomy for benign uterine diseases，recruited from the Department of Gynecology，West China Second University Hospital，Sichuan University，from January to June 2019 as study subjects.According to types of endometrial diseases，they were divided into EA group （n = 95），AH group （n= 25），and control group （n = 30）.Immunohistochemistry was used to measure DCLK1 protein expression in tissues of above three groups.To further assess the role of DCLK1，EA cell lines （HEC-1-A and Ishikawa）were transfected with DCLK1 overexpression plasmids or control vectors.These cells were grouped as the DCLK1 overexpression subgroup，and vector subgroup.And HEC-1-A and Ishikawa cell lines that were not transfected with plasmids were included into blank subgroup.The expression levels of DCLK1 and key proteins in AKT/m TOR signaling pathways of HEC-1-A and Ishikawa cells in the above three subgroups were detected.Ethical approval was granted by the Ethics Committee of West China Second University Hospital，Sichuan University （Approval No.2022-038），and all participants provided clinical research informed consent.

Results

①The proportion of abnormal uterine bleeding in patients of EA group，AH group，and control group were compared，and the difference was statistically significant （χ² =45.90，P ＜0.001）.The positive expression rates of DCLK1 in endometrial glands of these three groups of patients were 83.2%（79/95），76.0%（19/25），and 40.0%（12/30），respectively，and the difference was also statistically significant （χ²=21.83，P＜0.001）.②In the EA group，there was a significant difference in the positive expression rate of DCLK1 in patients with different pathological grades of EA tissue （χ²=14.70，P=0.003），and the expression level of DCLK1 was positively correlated with EA pathological grade （r_s=0.503，P ＜0.001）.③In HEC-1-A and Ishikawa cells of DCLK1 overexpression subgroup，DCLK1 protein expression levels were significantly higher than those in vector subgroup and blank subgroup，and protein kinase B （AKT），phosphorylated protein kinase B （p-AKT）and mammalian target of rapamycin （m TOR）protein expression levels were significantly higher than those in vector subgroup，and all the differences were statistically significant （P＜0.05）.

Conclusions

The expression level of DCLK1 is closely related to the malignant phenotype of EA，and it may also be related to the AKT/m TOR signaling pathway.DCLK1 may serve as a potential biomarker for the clinical diagnosis of EA patients and a therapeutic target.