Circulating MicroRNAs in gynecological malignancies: from detection to prediction.

MicroRNAs (miRNAs) have been demonstrated to play critical roles in many physiological and pathophysiological processes. The presence of altered miRNA profiles in human body fluids has been reported for a number of diseases including gynecological malignancies. In this review, we summarized the current progresses of circulating miRNAs associated with malignancies in gynecology, with an emphasizing on the circulating miRNAs as diagnostic and prognostic biomarkers in ovarian cancer, endometrial carcinoma and cervical cancer.

Differentially expressed microRNAs in the serum of cervical squamous cell carcinoma patients before and after surgery.

BACKGROUND: The purpose of this study was to detect the serum microRNAs (miRNAs) that are differentially expressed in cervical squamous cell carcinoma (SCC) patients and negative controls, with a focus on the miRNA profiles of the patients before and after surgery. The aim of the study is to evaluate the potential of these miRNAs as novel markers for the post-therapeutic monitoring of cervical SCC patients. RESULTS: A total of 765 serum miRNAs from 10 cervical SCC patients before surgery, 10 cervical SCC patients after surgery, and 10 negative controls were profiled using a TaqMan MicroRNA Array. A set of selected differentially expressed miRNAs were further analyzed in the patients at different perioperative periods, including preoperative, 1 week postoperative, and one month postoperative. The results showed that several serum miRNAs were differentially expressed in the cervical SCC patients compared with the negative controls, including miR-646, miR-141* and miR-542-3p. More importantly, we found that levels of specific serum miRNAs were deregulated in the pre- and postoperative stages, and these miRNAs could be useful for post-therapeutic monitoring of disease progression. Finally, we depicted a regulatory network of differentially expressed serum miRNAs, and many possible target genes were predicted in the estrogen-mediated signal pathways, supporting the hypothesis that cervical SCC is a hormone-associated gynecological disease. CONCLUSIONS: Our study demonstrated that the circulating miRNAs miR-646, miR-141* and miR-542-3p could potentially serve as non-invasive biomarkers for cervical SCC. The levels of these specific miRNAs might be useful for the post-therapeutic monitoring of disease progression. This is the first report showing that circulating miRNAs could serve as biomarkers for the therapeutic intervention of cervical SCC.

Circulating microRNAs identified in a genome-wide serum microRNA expression analysis as noninvasive biomarkers for endometriosis.

CONTEXT: There is currently no reliable noninvasive biomarker for the clinical diagnosis of endometriosis. Previous analyses have reported that circulating microRNAs (miRNAs) can serve as biomarkers for a number of diseases. OBJECTIVE: The study aims to detect the serum miRNAs that are differentially expressed between endometriosis patients and negative controls to evaluate the potential of these miRNAs as diagnostic markers for endometriosis. DESIGN: A total of 765 serum miRNAs were profiled using a TaqMan microRNA array in a pool of 10 endometriosis patients and a pool of 10 negative controls, and a set of selected miRNAs were further analyzed in a validation cohort consisting of sera from 60 patients and 25 controls including 10 samples used in array profiling. RESULTS: The relative expression levels of miR-199a and miR-122 were found to be up-regulated in endometriosis patient samples compared with control samples, whereas miR-145*, miR-141*, miR-542-3p, and miR-9* were down-regulated in endometriosis patients. Importantly, the relative expression of miR-199a (P < 0.05) and miR-122 can be used to discriminate between severe and mild endometriosis. We also found that miR-199a is well correlated with pelvic adhesion and lesion distribution (P < 0.05) and associated with hormone-mediated signaling pathways. Furthermore, we investigated the diagnostic value of these molecules and confirmed the optimal combination of miR-199a, miR-122, miR-145*, and miR-542-3p with area under the curve of 0.994 (95% confidence interval = 0.984-1.000, P < 0.001) and a cutoff point (0.4950) of 93.22% sensitivity and 96.00% specificity. CONCLUSIONS: Our study demonstrated that the circulating miRNAs miR-199a, miR-122, miR-145*, and miR-542-3p could potentially serve as noninvasive biomarkers for endometriosis. miR-199a may also play an important role in the progression of the disease. This is the first report that circulating miRNAs serve as biomarkers of endometriosis.

[Effects of levonorgestrel-releasing intrauterine system on endometrial estrogen and progesterone receptors in patients with endometrial hyperplasia].

OBJECTIVE: To investigate the changes in local endometrial contents of estrogen receptors (ER) and progesterone receptors (PR) after insertion of levonorgestrel-releasing intrauterine system (LNG-IUS) and evaluate the efficacy of LNG-IUS in treating endometrial hyperplasia. METHODS: The endometrial histological changes were observed in 25 anovulatory women with dysfunctional uterine bleeding after insertion of LNG-IUS, and the contents of ERs and PRs in the endometrium were measured by immunohistochemistry. RESULTS: The endometrial proliferation activity was obviously inhibited 6 months after LNG-IUS insertion with decreased endometrial glands, glandular dysplasia and decidualization of interstitial cells. The positive cell rate for ERs and PRs in the glandular epithelial and interstitial cells were significantly reduced after LNG-IUS insertion. CONCLUSIONS: LNG-IUS can reduce ER and PR expressions in the endometrium and inhibit endometrial proliferation, and therefore can be effective in treating simple and complex endometrial hyperplasia.

Expression of Bmi-1, P16, and CD44v6 in Uterine Cervical Carcinoma and Its Clinical Significance.

OBJECTIVE: Bmi-1, a putative proto-oncogene, is a core member of the polycomb gene family, which is expressed in many human tumors. The p16 protein negatively regulated cell proliferation, whereas CD44v6 is associated with proliferation as an important protein. Additionally, CD44v6 is an important nuclear antigen closely correlated to tumor metastasis. The present study aims to investigate the expression and significance of Bmi-1, p16, and CD44v6 in uterine cervical carcinoma (UCC). METHODS: A total of 62 UCC, 30 cervical neoplasic, and 20 normal cervical mucosal tissues were used in the current study. The expression of Bmi-1, p16, and CD44v6 in these tissues was determined using immunohistochemical assay. The relationships among the expression of these indices, the clinicopathologic features of UCC, and the survival rate of UCC patients were also discussed. The correlation between Bmi-1 protein expression and p16 or CD44v6 protein in UCC was analyzed. RESULTS: The expression of Bmi-1, p16, and CD44v6 was significantly high in cervical carcinoma compared with that in the cervical neoplasia and normal colorectal mucosa (P<0.05). The over-expression of Bmi-1 protein in UCC was apparently related to the distant metastasis (P<0.01) and the tumor, nodes and metastasis-classification, i.e. the TNM staging, World Health Organization (P<0.05). Nevertheless, the positive expression of p16 protein in UCC was not significantly associated with the clinicopathologic features (P>0.05). The Kaplan-Meier survival analysis showed that the over-expression of Bmi-1 significantly decreased the survival rate of UCC patients (P<0.05). A strong correlation indicated that there was statistical significance between the expression of Bmi-1 and CD44V6 proteins in UCC (r=0.419, P=0.001). CONCLUSIONS: The over-expression of Bmi-1 and CD44v6 protein closely correlate to the tumorigenesis, metastasis, and prognosis of UCC. Bmi-1 and CD44v6 may be used to predict the prognosis of cervical carcinoma. Bmi-1 may indirectly regulate the expression of CD44v6 in UCC patients. The positive expression of p16 protein is possibly associated with the tumorigenesis, but not with the metastasis or prognosis of UCC.

Clinical Significance of CENP-H Expression in Uterine Cervical Cancer.

OBJECTIVE: This work aims to investigate the expression pattern and clinicopathologic significance of centromere protein H (CENP-H) in uterine cervical cancer (UCC). METHODS: The level of CENP-H expression in the paraffin sections of 62 UCC cases was determined by the SP immunohistochemical method, with complete clinicopathologic data in all cases. Statistical analysis was conducted to evaluate the prognostic and diagnostic significance of CENP-H using SPSS13.0 software package. RESULTS: Immunohistochemical assay showed strong CENP-H expression in 61.29% (38/62) of the paraffin-embedded cervical cancer tissues. Statistical analysis revealed a strong correlation between the CENP-H expression and the clinical classification (P=0.038) of the cervical carcinoma. The expression increased with rise of the stages. The analysis of Cox proportional hazards regression model suggested that CENP-H expression (P=0.002) and tumor stage (P=0.001) were independent prognostic markers for the survival of UCC patients. The survival analysis showed that the survival rate was significantly lower in patients with high expression of CENP-H than in those with low expression of CENP-H (P=0.001). CONCLUSIONS: CENP-H is likely to be a valuable marker for carcinogenesis and progression of UCC. It might be used as the important diagnostic and prognostic marker for cervical carcinoma patients, especially for those at early stage.

Cumulus cells and their extracellular matrix affect the quality of the spermatozoa penetrating the cumulus mass.

OBJECTIVE: To investigate the role of the cumulus cells and the cumulus matrix in affecting the penetrability, morphology, acrosome reaction, and motility of human spermatozoa penetrating the cumulus oophorus. DESIGN: Controlled experimental laboratory study. SETTING: University gynecology unit. PATIENT(S): Women undergoing assisted reproduction treatment and men visiting the subfertility clinics. INTERVENTION(S): Human spermatozoa were allowed to penetrate through the cumulus oophorus and cell-depleted cumulus matrix in a capillary, and were treated with cumulus cell extract or hyaluronic acid. MAIN OUTCOME MEASURE(S): The morphology, acrosomal status, and motility of human spermatozoa were determined. RESULT(S): Fewer spermatozoa could penetrate the fresh cell-depleted matrix compared with intact cumulus oophorus. Spermatozoa that penetrated through the cumulus oophorus had higher percentages of normal morphology and acrosome reaction and had specific motility pattern. These effects were lost or reduced in the cell-depleted matrix that had been stored overnight. Hyaluronic acid, a main component of the cumulus matrix at concentration found in the cumulus oophorus, modulated sperm motility but did not affect spontaneous acrosome reaction. Cumulus cell extract did not affect sperm motility, but induced acrosome reaction. CONCLUSION(S): Both the cumulus matrix and the cumulus cells contribute to the effect of cumulus oophorus on spermatozoa penetrating through it.

Menopausal symptoms in mid-life women in southern China.

OBJECTIVES: To explore the prevalence of menopausal symptoms in Chinese women aged 40-65 years living in Guangdong province in southern China, and to investigate their care-seeking behavior. DESIGN: A cross-sectional population-based study performed in Guangdong province, PR China. METHODS: A total of 9939 women were selected by multistage cluster sampling. From November 2003 to July 2004, women were interviewed in person with a prepared questionnaire about symptoms experienced in the 2 months preceding the survey. The main outcome measurements were self-reported menopausal symptoms and related factors. RESULTS: The mean age of natural menopause was 48.9 years. The prevalence and severity of menopausal symptoms were low. The three most prevalent symptoms were insomnia, joint and muscle pain, and dizziness (in 37.2%, 35.7%, and 31.5% of the sample, respectively). Hot flushes were experienced by 17.5% of women. The factors associated with the frequency of menopausal symptoms included profession, education, type of menopause and the presence of physical or emotional problems. Ever and current hormone replacement therapy usage was reported in 0.8% and 1.3% of women, respectively. Of the total study population, 28.9% had sought health care because of menopausal symptoms. CONCLUSIONS: The prevalence of menopausal symptoms in southern Chinese women is low, and this is accompanied by low usage of hormone replacement therapy.

Cumulus cells reduce the spermatozoa-zona binding inhibitory activity of human follicular fluid.

OBJECTIVE: To investigate the effects of human follicular fluid cultured with cumulus cells to inhibit the binding of spermatozoa to the zona pellucida of oocytes. DESIGN: Controlled experimental laboratory study. SETTING: University gynecology unit. PATIENT(S): Women undergoing assisted reproduction program and men visiting the subfertility clinics. INTERVENTION(S): Culture medium and human follicular fluid were used to culture cumulus cells in vitro for specified time periods. MAIN OUTCOME MEASURE(S): Zona binding capacity and motility of spermatozoa after incubation with cumulus cells treated culture medium or human follicular fluid. RESULT(S): Compared with the control medium, spent culture media after culturing cumulus cells for 3, 5, and 7 hours did not affect the motility and zona binding capacity of the treated spermatozoa. Significantly more spermatozoa treated with human follicular fluid that had been preincubated with cumulus cells for 5 and 7 hours bound onto hemizona in hemizona binding assay when compared with those preincubated in human follicular fluid without cumulus treatment. The hemizona index increased with the increase in the duration of cumulus cell treatment. Human follicular fluid with or without cumulus cells maintained sperm motility to similar extent for 3 hours. CONCLUSION(S): Cumulus cells reduced the inhibitory effect of human follicular fluid on spermatozoa-zona binding in vitro in a time-dependent manner.