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

<b><i>Background/Aims:</i></b> The study aimed to evaluate molecular changes related to trophoblast adhesion in placenta accreta spectrum (PAS) disorders. <b><i>Methods:</i></b> 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. <b><i>Results:</i></b> Microarray analysis showed that 157 transcripts were &gt; 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], <i>p</i> &lt; 0.05). MMP19 showed moderate staining, with no difference between the adherent and non-adherent sites. ADAM28 and CTSS showed weak or no staining. <b><i>Discussion:</i></b> This limited study suggests that CTSV may be involved in the pathogenesis of PAS.

<b><i>研究背景与目的：</i></b> 本研究旨在探究胎盘植入谱系（placenta accreta spectrum, PAS）疾病中与滋养细胞黏附相关的分子变化。<b><i>研究方法：</i></b> 本研究于2010年4月至2013年3月间，对10例确诊胎盘植入谱系疾病的病例开展回顾性分析，所有病例均明确区分了滋养细胞黏附部位与非黏附部位。采用微阵列分析（microarray analysis）与逆转录聚合酶链反应（reverse transcription polymerase chain reaction, RT-PCR）鉴定黏附部位的上调基因，并通过免疫组织化学（immunohistochemistry）检测基因表达变化。<b><i>研究结果：</i></b> 微阵列分析显示，共有157个转录本的表达上调幅度超过3倍，包括：解整合素和金属蛋白酶28（a disintegrin and metalloproteinase-28, ADAM28，上调3.10倍）、组织蛋白酶V（cathepsin V, CTSV，上调3.73倍）、组织蛋白酶S（cathepsin S, CTSS，上调3.46倍）以及基质金属蛋白酶19（matrix metalloproteinase-19, MMP19，上调3.41倍）。RT-PCR检测显示相关基因的mRNA表达水平较高。免疫组织化学结果显示，非黏附部位的细胞外滋养层细胞（extravillous trophoblast, EVT）仅表现为微弱或无CTSV表达，而黏附部位侵入子宫肌层的EVT则呈现强阳性表达（组织学评分：中位数[最小值-最大值]，115.6[37.6–153.6] vs. 184.8[56.4–222.8]，<i>p</i> < 0.05）。MMP19染色呈中等强度，黏附部位与非黏附部位无显著差异。ADAM28与CTSS染色呈微弱或无表达。<b><i>研究讨论：</i></b> 本小样本研究提示，CTSV可能参与胎盘植入谱系疾病的发病过程。