CDC20 inhibitor Apcin inhibits embryo implantation in vivo and in vitro.

For successful implantation, endometrial receptivity must be established. The high expression of CDC20 in many kinds of malignant tumours has been reported, and it is related to the occurrence and development of tumours. According to these functions, we think that CDC20 may also play important roles in the process of embryo implantation. To prove our hypothesis, we observed the distribution and expression of CDC20 in mouse and human early pregnancy. The effect of E2 and/or P4 on the expression of CDC20 in human endometrial cells was detected by Western blot. To further explore whether CDC20 is an important factor in adhesion and proliferation. The results showed that the expression of CDC20 in the uterus and menstrual cycle of early pregnant mice was spatiotemporal. E2 can promote the expression of CDC20. On the contrary, P4 and E2 + P4 inhibited the expression of CDC20. We also detected the proliferation and adhesion of human endometrial cells. We found that the inhibition of CDC20 with its inhibitor Apcin could reduce the adhesion rate and proliferation ability to RL95-2 and HEC-1A cells, respectively. Inhibiting CDC20 by Apcin could interfere the embryo implantation of mouse. It is suggested that CDC20 may play an important role in the process of embryo implantation. SIGNIFICANCE OF THE STUDY: Embryo implantation is an extremely complex and delicate process, including identification, localisation, adhesion and invasion between embryo and endometrium. Studies have shown the process of embryo implantation is very similar to that of tumour invasion. CDC20 is a cancer-promoting factor. We found CDC20 is spatially and spatially expressed in mouse and human menstrual cycles and is regulated by oestrogen and progesterone. Apcin can inhibit the adhesion of JAR cells and embryo implantation of mouse. CDC20 may provide a new way to improve the success rate of assisted reproduction.

miR-501 is upregulated in cervical cancer and promotes cell proliferation, migration and invasion by targeting CYLD.

BACKGROUND: Cervical cancer is the common gynecological deadly malignancy worldwide. Here we attempted to evaluate the effects and mechanisms of microRNA-501-5p (miR-501) on the cell proliferation, migration, invasion and the clinical significance in the cervical cancer. METHODS: Cervical cancer HeLa cells were transfected with miR-501 mimic or inhibitor or siRNA against Cylindromatosis (CYLD) using Lipofectamine 2000. miR-501 expression was assessed in HeLa cells and cervical cancer specimens by real-time qRT-PCR. The functional roles of miR-501 were determined by CCK-8, colony formation, scratch wound healing and transwell assays. The apoptosis rate was detected by flow cytometry assay. CYLD, BCL-2, BAX, NF-κB p65 and phosphorylated p65 (p-p65) proteins were examined by Western blotting. CYLD expression was evaluated by immunohistochemistry in cervical cancer tissues. RESULTS: miR-501 was upregulated, whereas CYLD protein was downregulated in cervical cancer tissues compared to normal cervical tissues. miR-501 overexpression and CYLD protein downregulation were positively correlated with poor differentiation, tumor size, International Federation of Gynecology and Obstetrics (FIGO) stage and lymph node metastasis. CYLD was downregulated by miR-501 at both mRNA and protein levels in HeLa cells. miR-501 promoted cell proliferation, migration and invasion in cervical cancer, while inhibited the apoptosis. This is possibly due to the downregulation of CYLD and subsequent activation of NF-κB p65. CONCLUSIONS: miR-501 upregulation and CYLD downregulation are associated with the development and progression of cervical cancer. miR-501 promotes cervical cancer cell proliferation, migration and invasion possibly via downregulating CYLD and subsequently activating NF-κB p65. miR-501 might be a potential therapeutic target for cervical cancer.

Endometrial stromal sarcoma in combination with mixed type endometrial carcinomas: A case report and literature review.

RATIONALE: Endometrial stromal sarcoma (ESS) is rare, representing only approximately 0.2% of all uterine malignancies. Mixed type endometrial carcinomas (MT-ECs) are rare tumors with both type I and II features, and are difficult to diagnose. Cases of ESS and MT-ECs coexisting in the same patient are extremely rare. This study aimed to describe a case of ESS in combination with MT-ECs in a 47-year-old premenopausal woman. PATIENT CONCERNS: A woman presented to the hospital complaining of occasional abdominal pain and had high tumor markers: cancer antigen (CA) 19-9 (263.6 U/mL) and CA 125 (428.0 U/mL). Transvaginal ultrasound examination revealed a complex mass (12.3 × 9.1 × 6.3 cm) with solid and cystic components on the right rear wall of the uterus. Abdominopelvic computed tomography images showed a pelvic cystic-solid mixed mass. The patient underwent an exploratory midline laparotomy. The mass was hypothesized to be malignant on the uterine posterior wall. Tumor deposits were found on bilateral parametrium. On peritoneal implantation, multiple metastases were seen on the serosal surface of the bowel and greater omentum. A frozen section revealed a spindle cell sarcoma. DIAGNOSES: Pathological reports following surgery revealed concurrent ESS and MT-ECs. INTERVENTIONS: The patient underwent a total abdominal hysterectomy, bilateral salpingo-oophorectomy, total omentectomy, and macroscopic clearance of the tumor. Adjuvant chemotherapy was given. OUTCOMES: The patient was still alive when this report was written. LESSONS: Considering the rarity of ESS in combination with MT-ECs, this study presented an overview of the literature and discussed a number of histological and clinical issues. Nevertheless, etiology and pathogenesis of these tumors need further investigation.

[SiRNA inhibition of E6AP expression in cervical cancer cells].

OBJECTIVE: To study the effect of RNA interference (RNAi) targeting E6AP on the proliferation and apoptosis of HeLa cells. METHODS: HeLa cells were cultured and divided into 3 groups: blank control group, cells transfected with nonsense siRNA (small interference RNA), and cells transfected with specific E6AP siRNA. The expressions of E6AP mRNA and protein were detected by RT-PCR and Western blot before and after the transfection respectively. Cell proliferation was determined by methylthiazolyl tetrazolium (MTT). The cell apoptosis index was assessed by flow cytometry. RESULTS: Upon treatment with E6AP siRNA for 24, 48 and 72 h, the expression level of E6AP mRNA decreased 33%, 72% and 70% than siRNA treated group. The protein expression levels in 48 h and 72 h E6AP siRNA groups decreased 38%, 59% comparing with those of the nonsense siRNA treated group (P < 0.05). The proliferative capacity of cells transfectd with E6AP siRNA was significantly lower than blank control group (F = 101.38, P < 0.05) and siRNA treated group (F = 38.64, P < 0.05). The apoptosis index of HeLa cells treated with E6AP siRNA was significantly higher than that of the nonsense siRNA (F = 41.48, P < 0.05) and the blank control group (F = 86.36, P < 0.05). CONCLUSION: SiRNA targeting can effectively suppress the expression levels of E6AP mRNA, corresponding with a proliferation inhibition and an enhanced apoptosis of HeLa cells.

[The significance of K-Cl cotransport1 expression in cervical cancer tissues].

OBJECTIVE: To investigate the expression of K-Cl cotransport 1(KCC1) in cervical cancer tissues and to investigate its role in the onset and progression of cervical cancer. METHODS: Specimens of uterus were obtained from 60 patients aged 45.89 (25 approximately 73), 40 with cervical cancer, 10 with cervical intraepithelial neoplasia (CIN), and 10 with chronic cervicitis. Semi-quantitative RT-PCR was used to detect the mRNA expression. Western blotting was used to detect the protein expression of KCC1. Immunofluorescence assay was used to detect the location of KCC1 protein. RESULTS: The mRNA expression and protein expression of KCC1 were both significantly higher in the cervical cancer tissue than those in the CIN and chronic cervicitis tissues (all P < 0.05). The levels of KCC1 in the lowly differentiated cancer tissues (at grades G(2) and G(3)) were significantly higher than those in the highly differentiated cancer tissues (at grade G(1), P < 0.05). Western blotting revealed that the protein expression level of KCC1 was significantly higher in the cervical cancer tissues than in the CIN and chronic cervicitis tissues (both P < 0.05) and the protein expression levels of KCC1 in the cancer tissues at G" 2 and G(3) grades were significantly higher than that in the cancer tissues at grade G(1) (both P < 0.05). There were no significant differences in mRNA and protein expression between the early and terminal cervical cancer tissues. There were no significant differences in the mRNA and protein expression of KCC1 between the cervical cancer cases with or without lymph node metastasis. Immunofluorescence assay showed that KCC1 was located in the cellular membrane in all patients and the KCC1 expression level there was significantly higher in the cervical cancer tissues than in the tissues of CIN and chronic cervicitis. CONCLUSION: The expression of KCC1 is up-regulated in cervical cancer and it may play an important role in the onset and progression of cervical cancer.