Investigation of hub ferroptosis-related genes and the immune landscape in recurrent pregnancy loss and unexplained infertility using bioinformatics analysis.

Background: Recurrent pregnancy loss (RPL) and unexplained infertility (UI) are common pregnancy disorders that affect women's physical and mental health and lack effective treatment. Endometrial factors are one factor that leads to RPL. The latest research indicates that ferroptosis and immunity are closely related to the normal physiological function of the endometrium and may play a role in the pathogenesis of RPL and UI. Therefore, the present study analyzed the relationship between ferroptosis genes and immune infiltration in RPL and UI. Methods: We downloaded the GSE165004 dataset and analyzed differences in ferroptosis-related genes (FRGs) between RPL and UI patients and healthy controls. Hub differentially expressed ferroptosis-related genes (DE-FRGs) were screened using the LASSO algorithm, the SVM-RFE algorithm and the protein-protein interaction (PPI) network. Differences in immune infiltration between healthy endometrium and RPL and UI endometrium was analyzed, and the relationship between hub DE-FRGs and immune cell infiltration was examined. Results: We extracted 409 FRGs and identified 36 up-regulated and 32 down-regulated DE-FRGs in RPL and UI. Twenty-one genes were screened using the LASSO regression algorithm, and 17 genes were screened using the SVM-RFE algorithm. We intersected the LASSO genes, SVM-RFE genes and PPI network proteins to obtain 5 hub DE-FRGs. Gene set enrichment analysis (GSEA) functional enrichment analysis results indicated that the cytokine-cytokine receptor interaction signaling pathway was the common pathway for hub DE-FRGs. T follicular helper cells were highly infiltrated in RPL and UI, and M1 and M2 macrophages were highly infiltrated. The expression levels of MAPK1 and RELA positively correlated with T follicular helper cells. Conclusions: Ferroptosis-related genes may disrupt endometrial functions and signaling pathways and lead to the occurrence of RPL and UI.

Observation of the therapeutic effect of apatinib in advanced platinum-resistant recurrent epithelial ovarian cancer.

BACKGROUND: Apatinib is an oral anti-angiogenic drug that mainly targets vascular endothelial growth factor receptor 2 (VEGFR-2) and is widely used in a variety of solid tumours. The purpose of this study is to evaluate the clinical efficacy and safety of apatinib in patients with advanced platinum-resistant relapsed epithelial ovarian cancer (EOC). METHODS: A retrospective analysis was performed, the clinical data of patients with stage IIIC-IV platinum-resistant relapsed EOC between January 2014 and May 2018 were collected. The objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), and overall survival (OS) were reviewed and evaluated. The propensity score matching (PSM) method was used to determine the final case data included in this study. RESULTS: According to 1:2 propensity matching, 108 patients were finally taken into account: 36 in the apatinib group and 72 in the control group. The follow-up ended in January 2019, and the median follow-up time was 28 months. In the apatinib group, ORR was 30.56% and DCR was 66.67%, whereas in the control group, ORR was 16.67% and DCR was 44.44%. In the apatinib group, median PFS was 6.0 months (95% CI 3.69-8.31) and median OS was 15.8 months (95% CI 6.99-24.6), while in the control group, median PFS was 3.3 months (95% CI 2.44-4.16) and median OS was 9.2 months (95% CI 6.3-12.06); the difference was statistically significant (P < 0.05). Apatinib was more effective than conventional chemotherapy in reducing the risk of PFS [HR 0.40 (95% CI 0.22-0.76), P = 0.0017] and OS [HR 0.40 (95% CI 0.21-0.73), P = 0.002]. Multivariate Cox analysis showed that the course of treatment and decrease in serum CA125 levels are independent risk factors for PFS in patients, while apatinib, the length of treatment course and the location of the lesion are independent risk factors for recurrence affecting the OS of patients. The main grade 3-4 adverse events in the apatinib group were hypertension, hand-foot syndrome, and oral mucosal ulcers, and all adverse events were controllable. CONCLUSION: Apatinib was found to be both safe and effective in patients with advanced platinum-resistant relapsed EOC. More in-depth clinical research and applications should be carried out.

Application of carbon nanoparticles in laparoscopic sentinel lymph node detection in patients with early-stage cervical cancer.

OBJECTIVE: To investigate the value of carbon nanoparticles in identifying sentinel lymph nodes in early-stage cervical cancer. METHODS: From January 2014 to January 2016, 40 patients with cervical cancer stage IA2-IIA, based on the International Federation of Gynecology and Obstetrics (FIGO) 2009 criteria, were included in this study. The normal cervix around the tumor was injected with a total of 1 mL of carbon nanoparticles (CNP)at 3 and 9 o'clock. All patients then underwent laparoscopic pelvic lymph node dissection and radical hysterectomy. The black-dyed sentinel lymph nodes were removed for routine pathological examination and immunohistochemical staining. RESULTS: Among the 40 patients, 38 patients had at least one sentinel lymph node (SLN). The detection rate was 95% (38/40). One hundred seventy-three SLNs were detected with an average of 3.9 SLNs per side. 25 positive lymph nodes, which included 21 positive SLNs, were detected in 8 (20%) patients. Sentinel lymph nodes were localized in the obturator (47.97%), internal lilac (13.87%), external lilac (26.59%), parametrial (1.16%), and common iliac (8.67%) regions. The sensitivity of the SLN detection was 100% (5/5), the accuracy was 97.37% (37/38), and the negative predictive value was 100. 0% and the false negative rate was 0%. CONCLUSIONS: Sentinel lymph nodes can be used to accurately predict the pathological state of pelvic lymph nodes in early cervical cancer. The detection rates and accuracy of sentinel lymph node were high. Carbon nanoparticles can be used to trace the sentinel lymph node in early cervical cancer.

Overexpression of cysteine cathepsin L is a marker of invasion and metastasis in ovarian cancer.

Cysteine cathepsins (CTSs) are involved in the degradation and remodeling of the extracellular matrix and are associated with cellular transformation, differentiation, motility and adhesion in cancer development. Previous studies indicate that CTSs may be involved in ovarian cancer invasion and metastasis. However, due to the lack of large sample clinical studies and direct experimental evidence for the relationship between the expression of CTSs and invasion and metastasis, the diagnostic and prognostic value of CTSs in ovarian cancer progression has not been elucidated. In the present study, we observed that expression levels of CTSB, CTSL and CC in malignant ovarian tumors were significantly higher than the expression levels in benign tumors and normal ovarian tissues, yet their associations with clinicopathological features varied. In particular, CTSL was related to lymph node metastasis, CC was related to liver metastasis and omental metastasis, and CTSB and CTSL expression levels were found to be independent prognostic factors in ovarian cancer. Further study indicated that the serum level of CTSL was significantly higher in patients with ovarian malignant tumors than the levels in benign tumors and healthy controls, and the levels were elevated in low grade and advanced stage compared to the levels in high grade and early stage disease, suggesting that the serum level of CTSL may be a useful serum marker for the diagnosis of ovarian cancer. Furthermore, the expression of CTSL in ovarian cancer cells can greatly enhance the ability of cell invasion and metastasis, although no change was observed for cell adhesion. Taken together, we demonstrated that the overexpression of CTSL is involved in tumor invasion and metastasis, and the CTSL level in serum may be a marker for invasion and metastasis in ovarian cancer.

Overexpression of heparanase in ovarian cancer and its clinical significance.

It has been reported that heparanase (HPSE) is overexpressed in ovarian cancer and is associated with tumor invasion and metastasis. However, a systematic study on the contribution of HPSE to tumor metastasis is rarely reported. In this study, based on the measurement of HPSE serum concentration, the expression of HPSE at both the mRNA and protein levels in tumors and its effects on the biological behaviors of cancer cells, we elucidated the role of HPSE in tumor invasion and metastasis in ovarian cancer and concluded that either the expression of HPSE in cancer and/or the serum concentration of HPSE may be a useful biomarker for the evaluation of surgery effects and prognosis prediction.

Impact of post-operative hormone replacement therapy on life quality and prognosis in patients with ovarian malignancy.

The present study aimed to assess the impact of post-surgical hormone replacement therapy (HRT) on life quality and prognosis in women with ovarian malignancy. HRT (Premarin, Nilestriol and medroxyprogesterone) was administered following surgery in 31 patients with ovarian cancer. A total of 44 ovarian cancer patients of similar age, clinical stage and pathological features did not receive HRT following surgery. The expression of estrogen receptor (ER)-α, ERß and progesterone receptor (PR) in cancer tissues was detected by immunohistochemical staining. Serum levels of calcitonin (CT) and transforming growth factor (TGF)-α were determined by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. Data were analyzed using Kaplan-Meier survival curves, a log-rank test and a Cox scale risk model. Quality of life was assessed in the patient groups and in healthy post-menopausal women (control) based on a questionnaire developed by the European Organization of Research and Treatment of Cancer (EORTC-C30), as well as our own specific questionnaire. A log-rank test revealed no difference in survival between the patients with and without HRT (p>0.05), and a Cox model showed that HRT was not an independent prognostic factor. The accumulated survival rate did not differ significantly based on the expression of ERα, ERß or PR in patients with or without HRT (p>0.05). The serum TGFα levels prior to and following surgery were not significantly different in either of the two patient groups (p>0.05). Serum CT levels were higher in patients without HRT at 1.5 years following surgery (p<0.05), but no significant difference was found in the serum CT levels of patients receiving HRT. The HRT and non-HRT groups differed significantly with regard to the body and emotional functional sub-scales of the EORTC-C30 (p<0.05) and the sex quality and autonomic nerve maladjustment categories of our specific questionnaire (p<0.05). Findings of this study showed that HRT administered following surgery exhibited no apparent negative effect on prognosis in patients with ovarian cancer, regardless of ERα, ERß or PR expression in cancer tissues, and had no effect on serum transforming growth factor (TGF)-α levels. Post-surgical HRT aided in the stabilization of serum CT levels and improved the quality of life in these patients.

Clinical value of combined detection of serum matrix metalloproteinase-9, heparanase, and cathepsin for determining ovarian cancer invasion and metastasis.

AIM: This study evaluated the clinical value of the combined detection of serum cathepsin L (CL), heparanase (Hpa), and matrix metalloproteinase-9 (MMP-9) for determining the degree of ovarian cancer invasion and metastasis before surgery. PATIENTS AND METHODS: Enzyme-linked immunosorbent assays were used to measure the serum content of CL, Hpa, and MMP-9 in 217 patients with untreated ovarian cancer before surgery, 100 patients with benign ovarian tumors, and 101 healthy women as controls. In addition, the degrees of invasion and metastasis were assessed by the 'gold standard' of clinicopathological diagnosis. The associations of the preoperative serum CL, Hpa, and MMP-9 levels with the clinicopathological factors and metastatic status were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the usefulness of these markers for determining the degree of ovarian cancer invasion before surgery. RESULTS: The serum CL, Hpa, and MMP-9 levels were significantly higher (p=0.001) in patients with malignant ovarian cancer compared with patients with benign ovarian tumors and healthy controls. The serum CL level was significantly higher in patients with epithelial ovarian carcinoma compared with non-epithelial ovarian carcinoma (p=0.048), whereas the serum levels of Hpa (p=0.109) and MMP-9 (p=0.544) did not differ significantly between these two groups. The serum CL, Hpa, and MMP-9 levels correlated with the degree of differentiation and the FIGO staging (p>0.05). The serum CL (p=0.030) and MMP-9 (p=0.010) levels were significantly associated with peritoneal metastasis, and the serum Hpa level (p=0.042) was associated with distant metastasis. A ROC curve analysis revealed sensitivity of 60.9%, 69.6%, and 72.2%, and specificity of 57.4%, 67.2%, and 68.9% for the preoperative serum levels of CL, Hpa, and MMP-9, respectively, as tumor markers for the degree of extra-pelvic metastasis. CONCLUSION: Elevated serum CL, Hpa, and MMP-9 levels are correlated with malignant invasion and progression in ovarian cancer. The combined detection of serum CL, Hpa, and MMP-9 may be useful for determining the extent of ovarian cancer metastasis before surgery.

[Clinical study of autoantibody spectrum against ovarian cancer associated antigens combined with CA(125) in detecting and monitoring ovarian cancer].

OBJECTIVE: To evaluate the clinical value of autoantibody spectrum against ovarian cancer associated antigens combine CA(125) in detecting and monitoring ovarian cancer. METHODS: Circulating IgG, IgM autoantibodies against ovarian cancer associated antigens which included TM4SF1, C1D, TIZ, OV-142, FXR1 and OV-189 were measured by indirect ELISA in serum from 126 patients with ovarian cancer (prior treatment), 42 patients with benign ovarian masses, 142 healthy women. Cut off value of IgG, IgM autoantibodies were determined by receive operating characteristic (ROC) curve. CA(125) was measured in serum by immunoradiometric assay (IRMA). We evaluated the clinical value of combining multiple autoantibodies (autoantibody spectrum), combining autoantibody spectrum with CA(125) by binary logistic regresion. The positive ratio of autoantibody spectrum in serum (prior and post treatment) of 24 synchronization patients with ovarian cancer was analyzed to evaluate the value in monitoring state of illness. RESULTS: Our data indicated that serum contains IgG, IgM autoantibodies against ovarian cancer associated antigens. The positive ratio of IgG autoantibodies in serum from ovarian cancer patients and cancer-free patients were 34.1% - 47.6% and 13.0% - 19.0%, respectively (P < 0.05). The positive ratio of IgM autoantibodies in serum from ovarian cancer patients and cancer-free patients were 39.7% - 53.2% and 12.0% - 33.2%, respectively (P < 0.05). The positive ratio of IgG autoantibodies against FXR1 and IgM autoantibodies against TIZ, FXR1 and OV-189 in early stage (I-II) ovarian cancer (55.3%, 63.8%, 61.7% and 66.0%) were significantly higher than those in advanced (III-IV) ovarian cancer (34.2%, 39.2%, 26.6%, 45.6%; all P < 0.05). Combining five autoantibodies (TM4SF1 IgG, TM4SF1 IgM, C1D IgG, FXR1 IgG and TIZ IgM) showed significantly improved sensitivity (75.4%, P < 0.05), lower specificity (78.3%, P < 0.05) and similar accuracy (77.1%, P > 0.05) in detecting ovarian cancer compared to those of CA(125) (61.1%, 88.0%, 77.1%). But the autoantibody spectrum showed significantly improved sensitivity in classifying early stage (76.6%), compared to those of CA(125) (51.1%, P < 0.05). Combining autoantibody spectrum with CA(125) showed significantly improved sensitivity (85.7%), specificity (90.8%)and accuracy (88.7%) in detecting ovarian cancer compared to those of autoantibody spectrum alone (all P < 0.05), while CA(125) (61.1%, P < 0.05; 88.0%, P > 0.05; 77.1%, P < 0.05). The positive ratio of combine the autoantibody spectrum with CA(125) was significantly lower in 24 post-treatment serum (42%) compared to the pairing prior treatment serum (88%, P < 0.05). CONCLUSION: Combining the autoantibody spectrum against ovarian cancer associated antigens with CA(125) can improve sensitivity, specificity and accuracy in detecting early ovarian cancer and may be used to monitoring state of illness.

[Clinical evaluation of autoantibody of splice variant of BARD1 in detection of ovarian cancer].

OBJECTIVE: To investigate the value of autoantibody of breast cancer susceptibility 1-associated RING domain (BARD1) splice variant (OV-142) in detection of ovarian cancer. METHODS: We cloned OV-142 gene into plasmid pET-30b(+). The recombinant protein of OV-142 was expressed in pET-30b(+) system and purified. The autoantibody of OV-142 was detected by indirect enzyme-linked immunosorbent assay (ELISA). RESULTS: We successfully constructed the recombinant plasmid of OV-142. The recombinant protein was expressed in pET-30b(+) system and purified. The purification rate of the recombinant protein was up to 90%. The relative amount of autoantibody of OV-142 detected by indirect ELISA was analyzed by receiver operating characteristic curve (ROC) and the cutoff value was determined. Combination of the autoantibody IgG of OV-142 and CA(125) was analyzed by logistic regression. The sensitivity, specificity and accuracy was 71.4%, 89.1%, and 81.9%, respectively, which were higher than IgG (41.3%, 84.2%, 66.8%) and CA(125) (61.1%, 88.0%, 77.1%) when used alone each. CONCLUSIONS: OV-142 is a splice variant of BARD1. It may be a potential immunotherapy target of ovarian cancer. Detection of autoantibody of OV-142 is a potent complementary tool of CA(125) in ovarian cancer diagnosis.

[Relationship between hormone therapy in women with ovarian malignancy and prognosis].

OBJECTIVE: To explore the relationship between hormone therapy (HT) in women with ovarian malignancy and prognosis. METHODS: HT was used in 31 patients with ovarian cancer after surgery, and 44 cases with ovarian cancer served as control. The expression of estrogen receptor (ER)alpha, ERbeta and progesterone receptor (PR) was detected by immunohistochemical staining respectively. The level of serum calcitonin and transforming growth factor alpha (TGFalpha) was detected by radio-immune and enzyme-linked immunosorbent assay pre- or post-surgery, as well as half a year to one year later post-surgery respectively in these cases. The survival curve of Kaplan-Meier and log-rank test as well as scale risk of Cox model were used to analyze the relationship between HT and prognosis of ovarian cancer. RESULTS: (1) The results of log-rank test showed that there was no difference in survival curve of patients with or without HT [(1108 +/- 52), (1086 +/- 43) d; P = 0.940]; the results of scale risk of Cox model also showed that HT was not an independent prognosis factor for patients with HT. (2) There was no relationship with HT and the accumulated survival in patients with either positive or negative expression of ERalpha, ERbeta and PR in tissue; as well as between HT and the level of serum TGFalpha pre-, post-surgery, or half a year to one year after surgery. (3) The level of serum calcitonin in patients without HT post-surgery half a year to one year later was higher than that pre-surgery [(141 +/- 13), (95 +/- 11) microg/L; P < 0.05], but there was no significant difference between patients with HT half a year to one year later post-surgery and pre-surgery [(90 +/- 18) microg/L, (93 +/- 14) microg/L; P > 0.05]. (4) There was a significant difference in body and emotion function between HT and without HT groups [(1.84 +/- 1.50), (1.45 +/- 0.82); (12.69 +/- 10.20), (12.90 +/- 11.61); P < 0.05], as well as in sex quality and autonomic nerve maladjustment and in the special list made [(1.05 +/- 0.74), (1.77 +/- 1.08); (10.10 +/- 3.21), (13.09 +/- 4.30); P < 0.05]. CONCLUSIONS: There is no adverse influence on prognosis in using of HT for patients with ovarian cancer after surgery. HT for patients with ovarian cancer post-surgery can help keep a stable level of serum calcitonin as well as improve the quality of life.

[Screening and sero-immunoscreening of ovarian epithelial cancer associative antigens].

OBJECTIVE: To explore epithelial ovarian cancer (EOC) antigens that are potentially useful for cancer early detection and therapy. METHODS: A high quality cDNA library derived from ascites tumor cells of EOC patients (3 cases of serous EOC, 1 case of mucinous EOC, and 1 case of endometrial carcinoma of ovary) was constructed, and the method of combining serological analysis of recombinant cDNA expression libraries (SEREX) and suppression subtractive hybridization (SSH) was used for screening cDNA library. All of the positive clones were sequenced and bioinformatics analysis with BLAST software in GenBank was performed. Serological mini-arrays of recombinant tumor antigens (SMARTA) was used to investigate the prevalence of autoantibodies to these antigens in both 96 ovarian cancer patients and 96 cancer-free controls. RESULTS: Fifty-five positive clones encoding different antigenic genes of EOC recognized by IgG and (or) IgM were obtained. It showed that these 55 clones derived from 45 distinct genes and these genes could be grouped into 6 classes as following according to homology with known expressed sequence tag (EST): (1) known ovarian carcinoma related genes: BARD1, et al; (2) homologous genes with other tumors: TM4SF1, et al; (3) homologous genes with special tissues: ILF3, FXR1, et al; (4) homologous genes with special function: TIZ, C1D, et al; (5) embryo originating genes: PKHD1, et al; (6) novel genes: OV-189, et al. SMARTA results showed that the positive ratio of five EOC antigens TM4SF1 (28% vs. 9%), C1D (21% vs. 6%), BARD1 (23% vs. 5%), FXR1 (23% vs. 8%), OV-189 (31% vs. 13%) which reacting with their IgG autoantibodies, three antigens TIZ (26% vs. 8%), FXR1 (28% vs. 11%), and OV-189 (18% vs. 7%) which reacting with their IgM autoantibodies in patients was higher than in controls (P < 0.05). The positive ratio of EOC antigens FXR1 (34% vs. 16%), and OV-189 (46% vs. 23%) reacting with their IgG autoantibodies and TIZ (40% vs. 18%), FXR1 (46% vs. 18%) which reacting with their IgM autoantibodies in stage I-II patients was higher than that in stage III-IV (P < 0.05). The positive ratio of OV-189 (67% vs. 26%) which reacting with its IgG autoantibodies in well differentiated cases was higher than in moderately-poorly differentiated cases (P < 0.01). Combination of the above antigens showed a 66% sensitivity and 73% accuracy in discriminating EOC. Combination of the autoantibody profile of TM4SF1, C1D, TIZ, BARD1, FXR1, OV-189 with CA125 showed an 83% sensitivity and 80% accuracy in discriminating EOC. CONCLUSIONS: The strategy of combination of SEREX and SSH is a potent tool in isolating tumor-associated antigen genes. Autoantibody profile of TM4SF1, C1D, TIZ, BARD1, FXR1, OV-189 are potential tumor markers in EOC detection.