细胞核增殖抗原在子宫腺肌病子宫内膜-肌层交界区中的表达

The expression of PCNA in endometrial-myometrial interface of patients with adenomyosis

目的通过检测不同月经周期子宫内膜-肌层交界区(endometrial-myometrial interface,EMI)组织中细胞核增殖抗原(proliferating cell nuclear antigen,PCNA)的表达水平,探讨子宫腺肌病(adenomyosis, ADS)和非ADS子宫EMI区的增殖特点。方法选择2015年6月至2016年11月首都医科大学附属北京妇产医院因ADS行全子宫切除术患者的子宫标本30例为ADS组,同期因宫颈上皮内瘤变Ⅲ切除的子宫标本26例为对照组。取EMI区组织,采用免疫组化、qRT-PCR(quantificational real-time PCR)及western blot技术检测两组子宫EMI区组织中PCNA的表达情况。结果①PCNA表达的部位:内膜腺上皮细胞、间质细胞、血管内皮细胞及EMI区平滑肌细胞、异位内膜细胞中均可见PCNA的表达。②PCNA在ADS组中的表达:在ADS-EMI区组织中,PCNA mRNA持续性高表达,增殖期与分泌期比较,差异无统计学意义(P>0.05);PCNA蛋白表达亦失去周期性变化,相对灰度值:(1.06±0.29)、(0.98±0.19);在位内膜和异位内膜中,PCNA蛋白增殖期与分泌期的表达差异无统计学意义(P>0.05)。③PCNA在对照组中的表达:在EMI区中,PCNA mRNA表达随月经周期变化,增殖期是分泌期的2.06倍(P<0.05);PCNA蛋白在增殖期的表达亦显著高于分泌期(P<0.05),相对灰度值:(0.78±0.18)、(0.61±0.22)。在内膜组织中,增殖期PCNA的表达水平高于分泌期(P<0.05)。④两组间PCNA表达对比:与对照组EMI区相比,ADS-PCNA mRNA呈持续性高表达,失去周期性变化,增殖期mRNA表达水平是对照组的2.965倍,分泌期是对照组的5.794倍;同样,ADS-PCNA蛋白亦呈持续性高表达,失去周期性变化。但两组中,EMI区PCNA的表达水平均低于同组的内膜组织(P<0.05)。结论ADS子宫EMI区PCNA持续性高表达,失去周期性变化,造成交界区局部增生紊乱,这可能与ADS子宫不规则增厚密切相关,并且可能参与ADS的发生、发展。

Objective To detect the proliferating cell nuclear antigen(PCNA) expression of the endometrial-myometrial interface(EMI)in different menstrual periods in both adenomyosis(ADS) and non-ADS specimens to reveal the proliferation characteristics of EMI in both physiological and pathological state. Methods From June 2015 to November 2016, 30 cases of uterus specimens from patients undergoing hysterectomy for ADS in Beijing Obstetrics and Gynecology Hospital of Capital Medical University were selected as ADS group, and 26 cases of uterus specimens removed due to cervical intraepithelial neoplasia Ⅲ were set as non-ADS group. Immunohistochemistry, quantificational real-time PCR and western blot were used to detect the expression of PCNA in EMI.Results ① The location of PCNA: The expression of PCNA appeared in the endometrial glandular epithelial cells, the stromal cells, vascular endothelial cells, smooth muscle cells of the EMI region and the ectopic endometrium cells.② The expression of PCNA in the ADS group: The mRNA expression of PCNA in ADS EMI region was persistent higher and lost the periodic changes. The protein expression of PCNA had no significant difference in the proliferative phase and secretory phase,the relative gray value:(1.06±0.29) vs.(0.98±0.19). Besides, in both eutopic and ectopic endometrium, the expression of PCNA lost the periodic changes from the proliferative phase to the secretory one(P>0.05).③ The expression of PCNA in the non-ADS group: The mRNA expression of PCNA in the EMI region changed with the menstrual cycle. The proliferation-phase PCNA mRNA expression level was 2.06-fold higher(P<0.05). Meanwhile, the protein expression of PCNA changed with the menstrual cycle, which was higher in the proliferative phase(P<0.05);the relative gray value:(0.78 ± 0.18) vs.(0.61± 0.22). And in the endometrium, the expression of PCNA showed the similar pattern as in the EMI region(P<0.05).④ Differences in the expression of PCNA between the two groups: Compared with the non-ADS group, the mRNA expression of PCNA in ADS EMI region was persistent higher and lost the periodic changes. The PCNA mRNA expression in the proliferation phase was 2.965-fold higher than that in the non-ADS group, while in the secretory phase, the mRNA expression was 5.794 higher. The expression of PCNA protein showed the similar pattern.However, in both groups, the expression of PCNA in the endometrium was significantly higher than that in the EMI region(P<0.05).Conclusion In adenomyosis, the expression of PCNA in the EMI region shows persist high level, loses the cyclical changes, and leads to the proliferation dysfunction of the EMI region, which may relate to the irregular thickening of adenomyosis, and the occurrence and development of adenomyosis.