USP18 promotes cell proliferation and suppressed apoptosis in cervical cancer cells via activating AKT signaling pathway.

BACKGROUND: The deubiquitinating (DUB) enzyme ubiquitin-specific protease 18 (USP18), also known as UBP43, is an ubiquitin-specific protease linked to several human malignancies. However, USP18's underlying function in human cervical cancer remains unclear. In the current study, we aimed to analyse the role of USP18 and its signalling pathways in cervical cancer. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical staining were performed to analyse USP18 levels in cervical cancer and matched to adjacent normal tissues. Moreover, RNA interference (RNAi) and lentiviral-mediated vector transfections were performed to silence and overexpress USP18, respectively, in cervical cancer cells. Further, Cell Counting Kit-8 (CCK-8) and Annexin V/PI staining assays were used to assess its biological function in cell proliferation and apoptosis, respectively. A xenograft model was used to examine USP18's function in vivo. RESULTS: The present findings demonstrated that USP18 was overexpressed in cervical cancer specimens and cell lines. Silencing USP18 in SiHa and Caski cervical cancer cell lines inhibited cell proliferation, induced apoptosis, and promoted cleaved caspase-3 expression. In contrast, USP18 overexpression showed the opposite effects in human HcerEpic cells. A Gene Set Enrichment Analysis revealed that USP18 was enriched in the PI3K/AKT signalling pathway in cervical cancer. Hence, the PI3K/AKT inhibitor LY294002 was used to determine the relationship between USP18 and AKT in cervical cancer cells. Importantly, LY294002 significantly abolished the effects of USP18 overexpression in cervical cancer cells. In vivo, USP18 silencing inhibited human cervical cancer cells' tumorigenicity. CONCLUSIONS: The current study indicates that USP18 is an oncogenic gene in cervical cancer. Our findings not only deepened the understanding of USP18's biological function in cervical cancer pathogenesis, but we also provided novel insight for cervical cancer therapy. TRIAL REGISTRATION: Retrospectively registered.

miR-454-3p promotes proliferation and induces apoptosis in human cervical cancer cells by targeting TRIM3.

Abnormally expressed microRNAs have been demonstrated related to the development and progression of cervical cancer. However, the molecular mechanisms remain largely unkown. Here, we aimed to demonstrate the exact role of miR-454-3p in cervical cancer. Depletion of miR-454-3p in cervical cancer cells resulted in inhibition of cell growth and promotion of cell apoptosis. Bioinformatics analysis predicted that tripartite motif-containing 3 (TRIM3), a tumor suppressor gene in cervical cancer, is a promising target of miR-454-3p. Dual-luciferase reporter gene assay revealed that miR-454-3p directly target TIRM3 by binding to the 3'UTR of TIRM3. In cervical cancer cells (C-33A and SiHa) with endogenous low TRIM3 expression, decreased expression of miR-454-3p significantly elevated TRIM3 expression. In the cervical cancer cell (HeLa) with endogenous high TRIM3 expression, increased expression of miR-454-3p obviously inhibited TRIM3 expression and then manipulating cell growth and apoptosis, down-regulating the expression of P53 and cleaved caspase-3 via P38 MAPK signaling. Taken together, these findings demonstrated miR-454-3p as a cancer promoter by targeting TRIM3 in human cervical cancer.

Tripartite motif-containing protein 3 plays a role of tumor inhibitor in cervical cancer.

Many studies have been reported that tripartite motif-containing (TRIM) proteins play important roles in various cellular processes and involved in many diseases. TRIM3, a member of TRIM family, has been proved that it plays important roles in various cancers. Nevertheless, its effects on cervical cancer reminded unknown. This study aimed to explore TRIM3 function in cervical cancer cells. The results of quantitative real-time RT-PCR and western blotting showed that the TRIM3 expression was very low in five cervical cancer cell lines. The TRIM3 overexpression weakened cell viability, and promote apoptosis of C-33A and SiHa cells in vitro, and inhibit tumor growth in vivo, which suggested that TRIM3 could reduce proliferation of cervical cancer cells. Moreover, TRIM3 up-regulation enhanced caspase-3 activity and increased the expressions of cleaved caspase-3 and p53, at the same time decreased p38 phosphorylation level. In addition, TRIM3 down-regulation had opposite effects on cell proliferation and expressions of the three proteins, which could be suppressed by p38-specific inhibitor (SB203580). In conclusion, TRIM3 had the ability to suppress cell proliferation by inactivating p38 signaling pathway, which indicated that it might act as a tumor inhibitor and an underlying therapeutic target for cervical cancer.

Toll-like receptor 4 promotes proliferation and apoptosis resistance in human papillomavirus-related cervical cancer cells through the Toll-like receptor 4/nuclear factor-κB pathway.

Toll-like receptor 4 is overexpressed in various tumors, including cervical carcinoma. However, the role of Toll-like receptor 4 in cervical cancer remains controversial, and the underlying mechanisms are largely elusive. Therefore, Toll-like receptor 4 in cervical cancer and related mechanisms were investigated in this study. Quantitative reverse transcription polymerase chain reaction and western blot analyses were used to detect messenger RNA and protein levels in HeLa, Caski, and C33A cells with different treatments. Proliferation was quantified using Cell Counting Kit-8. Cell cycle distribution and apoptosis were assessed by flow cytometry. Higher levels of Toll-like receptor 4 expression were found in human papillomavirus-positive cells compared to human papillomavirus-negative cells. Proliferation of HeLa and Caski cells was promoted in lipopolysaccharide-stimulated groups but suppressed in short hairpin RNA-transfected groups. Apoptosis rates were lower in lipopolysaccharide-stimulated groups relative to short hairpin RNA-transfected groups. In addition, G2-phase distribution was enhanced when Toll-like receptor 4 was downregulated. Moreover, the pNF-κBp65 level was positively correlated with the Toll-like receptor 4 level in HeLa and Caski cells, though when an nuclear factor-κB inhibitor was applied to lipopolysaccharide-stimulated groups, the patterns of proliferation and apoptosis were opposite to those of the lipopolysaccharide-stimulated groups without inhibitor treatment. In conclusion, these data suggest that Toll-like receptor 4 promotes proliferation and apoptosis resistance in human papillomavirus-related cervical cancer cells at least in part through the Toll-like receptor 4/nuclear factor-κB pathway, which may be correlated with the occurrence and development of cervical carcinoma.

[Study on the expression and signification of TLR4/NO pathway in cervical tumorigenesis with high risk HPV infection].

OBJECTIVE: To explore the role of toll-like receptor (TLR)/ nitric oxide (NO) pathway in cervical tumor with high risk human papillomavirus (hrHPV) infection. METHODS: (1)Study was based on 36 women with nonmalignantcervical tissue as control group and 36 women with squamous cell cervical cancer (SCC), all with hrHPV infection which were assessed by using 14 types hrHPV E6/E7 mRNA real-time PCR kit. The amount of NO was detected by Griess reaction, the expression of inducible nitric oxide synthase(iNOS) was detected by immunohistochemistry (IHC). The mRNA expression of TLR3, TLR4, TLR7, TLR8, TLR9, nuclear factor-κB (NF-κB) p65 and iNOS in control and SCC epithelium which was captured by laser capture microdissection (LCM) were determined.(2)The expressions of TLR4 in CaSki, HeLa and C33a were detected by cell immunofluorescence method. The mRNA and protein expression of TLR/NO pathway transduction molecules including TLR4,NF-κBp65 and iNOS in CaSki, HeLa and C33a cell lines were detected by real-time PCR and western blot. RESULTS: (1)The level of NO was much higher in SCC group than that in control group [(42.92±0.36)µmol/L vs(15.49±0.24)µmol/L; P < 0.05 ]. iNOS was detected in 75% (27 cases ) of patients with squamous cervical carcinoma, while only 6% (2 cases) of normal controls were confirmed with positve result (P < 0.05). TLR/NO pathway maybe activated in SCC, for the mRNA levels of TLR3, TLR4, TLR7, TLR8, NF-κBp65 and iNOS increased significantly when compared to control group (all P < 0.05), and the greatest change in the expression level of TLR in SCC was spotted on TLR4(7.41±0.39 vs 1.86±0.21). (2)The results of immunofluorescence showed that TLR4 was located at plasma membrance of hrHPV positive HeLa and CaSki cells, while the integral optical density of TLR4 in HeLa cells (3 599±427) or CaSki cells (2 080±456) were higher than that in C33a cells (730± 96; P < 0.05). The mRNA and protein level of TLR4, NF-κBp65 and iNOS in HeLa and CaSki cells were higher than those of C33a cells (P < 0.05). CONCLUSION: TLR4/NO pathway is highly expressed in cervical cancer with hrHPV infection, while the pathway may be involved in cervical tumorigenesis with hrHPV infection.

[Retracted] Tripartite motif­containing 14 regulates cell proliferation and apoptosis in cervical cancer via the Akt signaling pathway.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that the GAPDH control western blotting data shown in Fig. 2B were strikingly similar to data appearing in different form in Fig. 1E in another article written by different authors at different research institutes [Liang T, Ye X, Liu Y, Qiu X, Li Z, Tian B and Yan D: FAM46B inhibits cell proliferation and cell cycle progression in prostate cancer through ubiquitination of ß­catenin. Exp Mol Med 50: 1­12, 2018]. Owing to the fact that the contentious data in the above article had already been published prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 22: 5145­5154, 2020; DOI: 10.3892/mmr.2020.11634].

YY1 is a novel potential therapeutic target for the treatment of HPV infection-induced cervical cancer by arsenic trioxide.

OBJECTIVE: YY1 is a zinc finger transcription factor involved in the regulation of cell growth, development, and differentiation. Although YY1 can regulate human papillomavirus-type (HPV) viral oncogenes E6 and E7, it remains unknown if YY1 plays a key role in carcinoma progression of HPV-infected cells. Here we sought to determine whether YY1 is upregulated in the cervical cancer tissues and YY1 inhibition contributes to apoptosis of cervical cancer cells, which is at least partly p53 dependent. Therefore, YY1 can be a potential therapeutic target for cervical cancer treatment by arsenic trioxide (As2O3). MATERIALS AND METHODS: The expression level of YY1 was examined and analyzed by Western blot in pathologically confirmed primary cervical cancer samples, in the adjacent normal samples, as well as in normal cervix samples. The effects of YY1 inhibition by specific small interfering RNA in HeLa cells were determined by Western blot analysis of p53 level, cell growth curve, colony formation assay, and apoptosis. The contribution of YY1 to As2O3-induced p53 activation and apoptosis was also examined by Western blot and cell cycle analysis. RESULTS: Here we report that the expression level of YY1 is significantly elevated in the primary cancer tissues. In HPV-positive HeLa cells, small interfering RNA-mediated YY1 inhibition induced apoptosis and increased the expression of p53. Treatment of HeLa cells with As2O3, a known anti-cervical cancer agent, reduced both protein and mRNA levels of YY1 in HeLa cells. YY1 knockdown significantly further enhanced As2O3-induced apoptosis. CONCLUSIONS: These results demonstrated that the expression of YY1 is upregulated in cervical carcinomas and that YY1 plays a critical role in the progression of HPV-positive cervical cancer. In addition, YY1 inhibition induces p53 activation and apoptosis in HPV-infected HeLa cells. Thus, YY1 is an As2O3 target and could serve as a potential drug sensitizer for anti-cervical cancer therapy.

[Expression of glyoxalase I and its effect on cell proliferation and apoptosis in endometrial carcinoma].

OBJECTIVE: To examine the expressions of glyoxalase I (GLO-I) in endometrial cancer tissues and cell lines and to investigate the roles of GLO-I on proliferation and apoptosis in endometrial cancer cells. METHODS: Immunohistochemistry, western blot and RT-PCR were used to investigate the expressions of GLO-I protein and mRNA in endometrial cancer tissues and Ishikawa cell lines;enzyme activity of GLO-I in normal endometrium, endometrial cancer and paraneoplastic tissue samples was detected with spectrophotometer;proliferation and apoptosis of Ishikawa cell before and after RNA interference (RNAi) procedure were detected by the methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. RESULTS: (1) There were significant differences of GLO-I expression between normal endometrium (0/19) and endometrial cancer tissues (76%, 22/29); these were also significant differences of enzyme activity of GLO-I among normal endometrium, paraneoplastic and endometrial cancer tissues (1.1, 0.8 vs 92.3 IU/mg; P < 0.01). Enzyme activity of GLO-I in fresh normal endometrium and paraneoplastic tissues was weak, while that of fresh endometrial cancer tissues was as high as 92.3 IU/mg in average. (2) The expression of GLO-I mRNA in Ishikawa cell transfected with GLO-I siRNA was significantly lower than that in negative group (0.25 ± 0.06 vs 0.93 ± 0.10, P < 0.01), and the similar results that in the expression of GLO-I protein (0.38 ± 0.06 vs 0.94 ± 0.13, P < 0.01). (3) Proliferation in Ishikawa cell was significantly inhibited after silencing RNA expression of GLO-I (P = 0.028). The apoptosis rate of cells transfected with GLO-I siRNA was significantly higher than that of negative control group and blank control group [(6.7 ± 0.8) % vs (1.2 ± 0.4)%, (1.4 ± 0.4)%; P < 0.01]. CONCLUSION: The expression and enzyme activity of GLO-I is significantly increased in endometrial cancer, which could promote abnormal proliferation and inhibit apoptosis in endometrial cancer cells.

Tripartite motif­containing 14 regulates cell proliferation and apoptosis in cervical cancer via the Akt signaling pathway.

Tripartite motif­containing (TRIM) 14 is a protein of the TRIM family. Studies have indicated that TRIM14 may be used as an oncogene in tumor cells, such as osteosarcoma, non­small cell lung cancer and breast cancer through different pathways. However, the functions of TRIM14 in cervical cancer cells remain unclear. Therefore, this study aimed to investigate the functions of TRIM14 in cervical cancer cells and its underlying mechanism. Caski cells stably expressing TRIM14 and SiHa, and HeLa cells stably expressing TRIM14 short hairpin RNA were constructed by lentivirus­mediated overexpression or knockdown systems. The effects of TRIM14 on proliferation and apoptosis of cervical cancer cells were detected by Cell Counting Kit­8 (CCK­8) assay and flow cytometry, respectively. In addition, reverse transcription­quantitative (RT­q) PCR and western blotting were used to investigate the expression levels of TRIM14 and of signaling pathway marker protein including P21, caspase­3, cleaved caspase­3, Akt and phosphorylated Akt. The results of RT­qPCR and western blotting revealed that TRIM14 was highly expressed in human cervical cancer tissues and cell lines compared with adjacent normal tissues and normal cervical epithelial cells. TRIM14 also regulated cell proliferation and apoptosis of human SiHa, HeLa and Caski cervical cancer cell lines through the Akt signaling pathway. Additionally, TRIM14 protein levels were related to the clinical and pathological features of cervical cancer. CCK­8 assay and flow cytometry demonstrated that TRIM14 expression could promote cervical cancer cell proliferation and autophagy suppression. Taken together, TRIM14­induced cell proliferation and apoptosis inhibition may by evoked by the activation of the Akt pathway. This study demonstrated the role of TRIM14 in cervical cancer, and reveals its mechanism of action as a potential therapeutic target for cervical cancer.

ARHGAP17 suppresses tumor progression and up-regulates P21 and P27 expression via inhibiting PI3K/AKT signaling pathway in cervical cancer.

ARHGAP17 has long been thought to be involved in the maintenance of tight junction and epithelial barrier. Recently, a few Rho GTPase activating proteins (RhoGAPs) have been identified as tumor suppressors in some human cancers. The present study aimed to explore ARHGAP17 expression in cervical cancer and the possible function in tumor progression. ARHGAP17 expression in cervical cancer cell lines was assessed by RT-PCR and Western blotting. ARHGAP17 expression in cervical cancer tissues and normal tissues was assessed by immunohistochemistry. The cell proliferation was determined using CCK-8 in vitro and subcutaneous xenograft model in vivo. Here, we showed lower expression of ARHGAP17 in cell lines and human cervical cancer samples. Manipulation of ARHGAP17 affected cell proliferation in vitro and tumor growth in vivo. Furthermore, the phosphorylation of AKT was enhanced in ARHGAP17 silencing cervical cancer cells. ARHGAP17 can elevate P21 and P27 expression level through inhibiting PI3K/AKT signaling pathway. Stepwise investigations demonstrated that ARHGAP17 suppressed malignant phenotype of cervical cancer cells via inhibiting PI3K/AKT signaling pathway. These results reveal that ARHGAP17 functions as a tumor suppressor in cervical cancer that suppresses tumor growth, at least partly, through inhibition of PI3K/AKT signaling and up-regulation of P21 and P27 expression.

Identification of reliable biomarkers of human papillomavirus 16 methylation in cervical lesions based on integration status using high-resolution melting analysis.

Background: The dynamic methylation of human papillomavirus (HPV) 16 DNA is thought to be associated with the progression of cervical lesions. Previous studies that did not consider the physical status of HPV 16 may have incorrectly mapped HPV 16 methylomes. In order to identify reliable biomarkers for squamous cervical cancer (SCC), we comprehensively evaluated the methylation of HPV 16 depending on the integration incidence of each sample. Methods: Based on the integration status of 115 HPV 16-infected patients (50 SCC, 30 high-grade squamous intraepithelial lesion [HSIL], and 35 low-grade squamous intraepithelial lesion [LSIL]) and HPV 16-infected Caski cell lines by PCR detection of integrated papillomavirus sequences, we designed a series of primers that would not be influenced by breakpoints for a high-resolution melting (HRM) PCR method to detect the genome methylation. Results: A few regions with recurrent interruptions were identified in E1, E2/E4, L1, and L2 despite scattering of breakpoints throughout all eight genes of HPV 16. Frequent integration sites often occurred concomitantly with methylated CpG sites. The HRM PCR method showed 100% agreement with pyrosequencing when 3% was set as the cutoff value. A panel of CpG sites such as nt5606, nt5609, nt5615, and nt5378 can be combined in reweighing calculations to distinguish SCC from HSIL and LSIL patients which have high sensitivity and specificity (88% and 92.31%, respectively). Conclusions: Our research shows that combination of CpG sites nt5606, nt5609, nt5615, and nt5378 can be used as potential diagnosis biomarkers for SCC, and the HRM PCR method is suitable for clinical methylation analysis.

[The clinical application of spectral karyotyping in the analysis of chromosomal abnormalities].

OBJECTIVE: To evaluate value of spectral karyotyping (SKY) in the detection of chromosomal abnormalities. METHODS: A total of 17 metaphase chromosome samples were investigated by SKY, including 10 normal and 5 balanced translocation samples of peripheral blood lymphocytes, one der(Y) sample of peripheral blood lymphocytes and one marker chromosome sample of amniotic fluid cells. The results were compared with those of G-banding diagnosis. RESULTS: Ten normal and 5 balanced translocation samples were diagnosed successfully by SKY in accordance with the results of G-banding; furthermore, SKY analysis revealed that the der(Y) fragment originated from p-arm of chromosome 21 while the marker chromosome originated from chromosome 5. CONCLUSION: SKY is a very sensitive and specific whole genome analysis tool for chromosomal abnormality diagnosis, and exceedingly valuable in the diagnosis on complex chromosomal abnormalities that can not be determined by G-banding.

Involvement of FAK-ERK2 signaling pathway in CKAP2-induced proliferation and motility in cervical carcinoma cell lines.

Cervical carcinoma is the fourth most common cause of death in woman, caused by human papillomavirus (HPV) infections and arising from the cervix. Cytoskeleton-associated protein 2 (CKAP2), also known as tumor-associated microtubule-associated protein, has been linked to tumorigenic effects. In the present study, we screened CKAP2 as a new candidate gene which promotes development of cervical carcinoma, in two independent datasets (TCGA and GSE27678). Results showed that CKAP2 expression was significantly up-regulated in cervical cancerous tissues compared with normal counterparts. Gene set enrichment analysis (GSEA) showed that metastasis, cell cycle and FAK pathways were related with elevated CKAP2 expression. Knockdown of CKAP2 expression significantly inhibited cell proliferation, migration and invasion both in HeLa and C-33A cells. And depletion of CKAP2 down-regulated the expression of metastasis and cell cycle related proteins as well as the phosphorylation of ERK2 (p-ERK2), except E-cadherin. In vivo experiment revealed that knockdown of CKAP2 inhibited C-33A cells proliferation. However, FAK inhibitor PF-562271 and ERK2 inhibitor VX-11e treatment significantly inhibited CKAP2 overexpression-induced cell proliferation, migration and invasion in SiHa cells. In conclusion, our study suggests that CKAP2 acts as a functional oncogene in cervical carcinoma development and may exert its function by targeting FAK-ERK2 signaling pathway.

Fertility-conservation combined therapy with hysteroscopic resection and oral progesterone for local early stage endometrial carcinoma in young women.

OBJECTS: This paper explored the suitable population for the combined therapy of hysteroscopic resection and oral megestrol acetate (MA) to treat local stage I endometrial cancer. Therapeutic effectiveness, safety, as well as pregnancy rate and relapse rate after treatment were also examined. The aim was to provide guidance for the treating similar cases in the future. METHODS: This perspective study analyzed the clinical data of early stage endometrial cancer patients who have received combined therapy of hysteroscopic resection of local endometrial lesion and oral administration of MA at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai. RESULTS: A total of six patients met the entry criteria and were enrolled into the trial. All of them achieved a pathologic complete response to hysteroscopic resection of local lesion combined with oral administration of MA for 3 months to 6 months. Among the patients, three became pregnant after natural conception and had healthy infants delivered vaginally at full term without assistance. No relapse occurred in the follow-up study over 48.5 months on average. CONCLUSIONS: In early-stage endometrial cancer, young patients who had already given birth demand may receive hysteroscopic resection combined with oral administration of MA as conservative treatment. The patients can consider natural conception after complete remission, but a close follow-up was crucial to ensuring that the patients were free from other factors affecting childbearing ability.

Effects of tissue factor pathway inhibitor-2 expression on biological behavior of BeWo and JEG-3 cell lines.

OBJECTIVES: To investigate the effect of tissue factor pathway inhibitor-2 (TFPI-2) expression on biological behavior of BeWo and JEG-3 cell lines. MATERIAL AND METHODS: The expression of TFPI-2 in BeWo and JEG-3 cells was upregulated by pEGFP-N3-TFPI-2 and downregulated by small interference RNA transfection, confirmed by Western blotting assay and real-time polymerase chain reaction (RT-PCR). Boyden chamber, Cell Counting Kit-8 (CCK-8), and Hoechst 33258/terminal deoxynucleotidyltransferase-mediated UTP end labeling (TUNEL) assays were used for migration, invasion, and proliferation/apoptosis analysis, respectively. RESULTS: In Western blotting and RT-PCR assay, protein and messenger RNA (mRNA) expression of TFPI-2 in transfected BeWo and JEG-3 cells were confirmed. Expression of TFPI-2 inhibited BeWo and downregulated JEG-3 cell migration, invasion, proliferation, and induced apoptosis (P < .05) in Boyden chamber, CCK-8, Hoechst 33258, and TUNEL detection, respectively. CONCLUSIONS: TFPI-2 expression caused invasion and proliferation impair and induced apoptosis in TFPI-2 regulated BeWo and JEG-3 cells. It provides a clue for potential role of TFPI-2 in trophoblast.

Changes of plasma and placental tissue factor pathway inhibitor-2 in women with preeclampsia and normal pregnancy.

OBJECTIVES: To investigate the maternal and fetal plasma changes of tissue factor pathway inibitor-2 (TFPI-2) and its placental expression in women with normal pregnancy and preeclampsia. MATERIAL AND METHODS: We assessed the plasma TFPI-2 level in non-pregnant, normal pregnant and postpartum women, detected fetal plasma level and expression in placenta, and compared the changes in women with preeclampsia. Time-resolved fluoroimmunoassay and immunohistochemistry were used for plasma and placenta tissue detection, respectively. RESULTS: Maternal plasma levels of TFPI-2 in normal pregnant women at 13weeks of gestation increased 9.3 times as compared with healthy non-pregnant women (149.3+/-17.1 versus 16.0+/-3.6ng/ml)reached a maximum level ( 282.6+/-17.1ng/ml) at 39weeks of gestation, and dramatically decreased to nearly a non-pregnant level on the first day of postpartum (32.3+/-7.1ng/ml); similar change was found in the placental expression. Fetal plasma TFPI-2 was significantly lower than the maternal level at delivery. The maternal plasma TFPI-2 in preeclampsia was significantly lower compared with that in normal pregnancy, coupled with significantly higher placental expression. CONCLUSIONS: Placenta may be the main site of the high level of TFPI-2 production in maternal circulation, and the dramatic changes in preeclampsia provide a clue to elucidate its pathogenesis.