Supplementary Material for: Possible Association between Cathepsin V and the Development of Placenta Accreta Spectrum Disorders

Background/Aims: The study aimed to evaluate molecular changes related to trophoblast adhesion in placenta accreta spectrum (PAS) disorders. Methods: A retrospective analysis of 10 PAS cases in which both the trophoblast adherent site and the non-adherent site were identified was performed in April 2010 and March 2013. Microarray analysis and reverse transcription polymerase chain reaction (RT-PCR) analyses were performed to extract upregulated genes in the adherent site. Gene expression changes were examined by immunohistochemistry. Results: Microarray analysis showed that 157 transcripts were > 3-fold upregulated, including the following: a disintegrin and metalloproteinase-28 (ADAM28), 3.10-fold; cathepsin V (CTSV), 3.73-fold; cathepsin S (CTSS), 3.46-fold; and matrix metalloproteinase-19 (MMP19), 3.41-fold. RT-PCR showed relatively high mRNA expressions. On immunohistochemistry, extravillous trophoblast (EVT) at the non-adherent site showed weak or no CTSV expression, whereas EVT that invaded myometrium at the adherent site showed strong expression (histological score, median [min-max], 115.6 [37.6–153.6] vs. 184.8 [56.4–222.8], p < 0.05). MMP19 showed moderate staining, with no difference between the adherent and non-adherent sites. ADAM28 and CTSS showed weak or no staining. Discussion: This limited study suggests that CTSV may be involved in the pathogenesis of PAS.

背景与目的：本研究旨在探讨胎盘植入谱系（placenta accreta spectrum, PAS）疾病中与滋养细胞黏附相关的分子变化。
方法：本研究于2010年4月至2013年3月期间，纳入10例经确认同时存在滋养细胞黏附部位与非黏附部位的PAS病例，开展回顾性分析。采用基因芯片分析与逆转录聚合酶链反应（reverse transcription polymerase chain reaction, RT-PCR）技术，筛选黏附部位的上调基因，并通过免疫组织化学法检测基因表达变化。
结果：基因芯片分析显示，共有157个转录本的表达上调幅度超过3倍，包括：解整合素金属蛋白酶28（ADAM28，上调3.10倍）、组织蛋白酶V（CTSV，上调3.73倍）、组织蛋白酶S（CTSS，上调3.46倍）以及基质金属蛋白酶19（MMP19，上调3.41倍）。RT-PCR检测结果显示，上述基因的mRNA表达水平相对较高。免疫组化结果显示，非黏附部位的绒毛外滋养层细胞（extravillous trophoblast, EVT）中CTSV表达微弱或呈阴性；而黏附部位侵袭子宫肌层的EVT则呈强阳性表达（组织学评分：中位数[最小值-最大值]分别为115.6[37.6~153.6]与184.8[56.4~222.8]，p<0.05）。MMP19染色呈中等强度，黏附部位与非黏附部位的染色强度无显著差异。ADAM28与CTSS的染色均呈微弱或阴性表达。
讨论：本项小样本研究提示，CTSV可能参与了PAS的发病过程。