Background
MicroRNAs are small noncoding RNAs with important regulatory functions. Although well-studied
in cancer, little is known about the role of microRNAs in premalignant disease. Complete
hydatidiform moles are benign forms of gestational trophoblastic disease that progress
to gestational trophoblastic neoplasia in up to 20% of cases; however, there is no
well-established biomarker that can predict the development of gestational trophoblastic
neoplasia.
Objective
This study aimed to investigate possible differences in microRNA expression between
complete moles progressing to gestational trophoblastic neoplasia and those regressing
after surgical evacuation.
Study Design
Total RNA was extracted from fresh frozen tissues from 39 complete moles collected
at the time of uterine evacuation in Brazil. In the study, 39 cases achieved human
chorionic gonadotropin normalization without further therapy, and 9 cases developed
gestational trophoblastic neoplasia requiring chemotherapy. Total RNA was also extracted
from 2 choriocarcinoma cell lines, JEG-3 and JAR, and an immortalized normal placenta
cell line, 3A-subE. MicroRNA expression in all samples was quantified using microRNA
sequencing. Hits from the sequencing data were validated using a quantitative probe-based
assay. Significantly altered microRNAs were then subjected to target prediction and
gene ontology analyses to search for alterations in key signaling pathways. Expression
of potential microRNA targets was assessed by quantitative real-time polymerase chain
reaction and western blot. Finally, potential prognostic protein biomarkers were validated
in an independent set of formalin-fixed paraffin-embedded patient samples from the
United States (15 complete moles progressing to gestational trophoblastic neoplasia
and 12 that spontaneously regressed) using quantitative immunohistochemistry.
Results
In total, 462 microRNAs were identified in all samples at a threshold of <1 tag per
million. MicroRNA sequencing revealed a distinct set of microRNAs associated with
gestational trophoblastic neoplasia. Gene ontology analysis of the most altered transcripts
showed that the leading pathway was related to response to ischemia (P<.001). Here, 2 of the top 3 most significantly altered microRNAs were mir-181b-5p
(1.65-fold; adjusted P=.014) and mir-181d-5p (1.85-fold; adjusted P=.014), both of which have been shown to regulate expression of BCL2. By quantitative real-time polymerase chain reaction, BCL2 messenger RNA expression was significantly lower in the complete moles progressing
to gestational trophoblastic neoplasia than the regressing complete moles (−4.69-fold;
P=.018). Reduced expression of BCL2 was confirmed in tissue samples by western blot.
Immunohistochemistry in the independent patient samples revealed significantly lower
cytoplasmic expression of BCL2 in the villous trophoblasts from cases destined for
progression to gestational trophoblastic neoplasia compared with those that regressed,
both with respect to staining intensity (optic density 0.110±0.102 vs 0.212±0.036;
P<.001) and to the percentage of positive cells (16%±28% vs 49.4%±28.05%; P=.003).
Conclusion
Complete moles progressing to gestational trophoblastic neoplasia are associated with
a distinct microRNA profile. miR-181 family members and BCL2 may be prognostic biomarkers
for predicting gestational trophoblastic neoplasia risk.
Key words
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Article info
Publication history
Published online: October 05, 2020
Accepted:
September 29,
2020
Received in revised form:
September 24,
2020
Received:
July 20,
2020
Footnotes
L.H.L. is currently affiliated to the Department of Pathology, New York University Langone Health, New York, NY.
The authors report no conflict of interest.
This study received financial supported from the Donald P. Goldstein, MD, Trophoblastic Tumor Registry Endowment; Dyett Family Trophoblastic Disease Research and Registry Endowment; Aperfeiçoamento de Pessoal de Nível Superior (Research Networks under Universidade Estadual Paulista’s Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)-PrInt PROJECT; finance code 001 [I.M.]); and São Paulo Research Foundation (Fundação de Amparo à Pesquisa do Estado de São Paulo [FAPESP]) (grant number 2020/08830-6). The funding agencies had no direct role in the generation of the data or the manuscript. The Donald P. Goldstein, MD, Trophoblastic Tumor Registry Endowment and the Dyett Family Trophoblastic Disease Research and Registry Endowment funded the transportation, maintenance of samples, and resources for the laboratory techniques. CAPES and FAPESP helped to maintain the international collaboration between the institutions through the researchers’ exchange program.
Cite this article as: Lin LH, Maestá I, St. Laurent JD, et al. Distinct microRNA profiles for complete hydatidiform moles at risk for malignant progression. Am J Obstet Gynecol 2021;224:372.e1-30.
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