Comprehensive analysis of hub genes associated with cisplatin-resistance in ovarian cancer and screening of therapeutic drugs through bioinformatics and experimental validation.

BACKGROUND: To identify key genes associated with cisplatin resistance in ovarian cancer, a comprehensive analysis was conducted on three datasets from the GEO database and through experimental validation. METHODS: Gene expression profiles were retrieved from the GEO database. DEGs were identified by comparing gene expression profiles between cisplatin-sensitive and resistant ovarian cancer cell lines. The identified genes were further subjected to GO, KEGG, and PPI network analysis. Potential inhibitors of key genes were identified through methods such as LibDock nuclear molecular docking. In vitro assays and RT-qPCR were performed to assess the expression levels of key genes in ovarian cancer cell lines. The sensitivity of cells to chemotherapy and proliferation of key gene knockout cells were evaluated through CCK8 and Clonogenic assays. RESULTS: Results showed that 12 genes influenced the chemosensitivity of the ovarian cancer cell line SKOV3, and 9 genes were associated with the prognosis and survival outcomes of ovarian cancer patients. RT-qPCR results revealed NDRG1, CYBRD1, MT2A, CNIH3, DPYSL3, and CARMIL1 were upregulated, whereas ERBB4, ANK3, B2M, LRRTM4, EYA4, and SLIT2 were downregulated in cisplatin-resistant cell lines. NDRG1, CYBRD1, and DPYSL3 knock-down significantly inhibited the proliferation of cisplatin-resistant cell line SKOV3. Finally, photofrin, a small-molecule compound targeting CYBRD1, was identified. CONCLUSION: This study reveals changes in the expression level of some genes associated with cisplatin-resistant ovarian cancer. In addition, a new small molecule compound was identified for the treatment of cisplatin-resistant ovarian cancer.

Preparation, characterization, and coating effect of bio-active nano-emulsion based on combined plant essential oils on quality of grass carp fillets.

This study aimed to develop a satisfactory essential oil (EO) nano-emulsion through high pressure microjet technology and explore its physiochemical properties and synergistic coating effects on grass carp fillets. The optimal conditions for oregano/litsea cubeba (6:4, wt%/wt%) nano-emulsion were shown to be 80 s high pressure microjet pretreatment time, 9000 lb per square inch pretreatment pressure, 6 % oil phase, and 3:2 Km (mass ratio of surfactant to co-surfactant). The obtained nano-emulsion exhibited 100.42 ± 0.96 nm oil diameter at 4 °C after 15-day storage, coupled with high stability after centrifugation, freeze-thaw and heating treatment. Compared with untreated samples at day 6 storage, the nano-emulsion-treated grass carp fillets exhibited improved textural properties, higher water-holding capacity (74.23 % ± 0.80 %), lower total volatile basic nitrogen (TVB-N, 13.46 ± 0.30 mg/100g)/thiobaric acid (TBA,0.43 ± 0.02 mgMDA/100g), and lower total viable spoilage bacteria count (4.98 ± 0.21 lgCFU/g). This study facilitates understanding the combined EOs nano-emulsion on improving the shelf life of grass carp fillets.

Non-invasive Amino Acid Profiling of Embryo Culture Medium Using HPLC Correlates With Embryo Implantation Potential in Women Undergoing in vitro Fertilization.

This study aimed to determine the correlation between amino acid profiling of a 3-day-old embryo culture medium and embryo implantation potential in women undergoing in vitro fertilization (IVF). The data of 98 patients who received IVF treatment in our hospital from December 2015 to February 2017 were retrospectively analyzed. The 98 patients were grouped into a pregnant group (gemellary pregnancy), a non-pregnant group (non-pregnancy), and a blank control group. The amino acids from a 3-day-old embryo culture medium and blank control medium were collected and were analyzed using high performance liquid chromatography (HPLC). The HPLC results showed that amino acids including aspartate (ASP), serine (SER), glycine (GLY), histidine (HIS), taurine (TAU), arginine (ARG), threonine (THR), alanine (ALA), and proline (PRO) were detected in the 3-day-old embryo culture medium and blank control medium. There are significant differences between the pregnant group and non-pregnant group in peak height (H)-SER, surface area (S)-ASP, S-SER, S-HIS, and S-ALA. The discrimination analysis according to the peak height and peak area of amino acids revealed that the prediction rate of the pregnant group, non-pregnant group, and blank control group were 82.7, 95.7, and 100%. Further, by using the principal component analysis, we found that the prediction rate in these three groups were 90.4, 91.3, and 100%. Our data may suggest that using amino acid concentrations for principal component analysis and discriminant analysis has high accuracy in predicting the relationship between amino acid fingerprint and embryo implantation potential.

MiR-210-3p protects endometriotic cells from oxidative stress-induced cell cycle arrest by targeting BARD1.

Endometriosis is associated with benign but adversely developed cysts in the extrauterine environment. The oxidative imbalanced environment induces DNA damage and affects cell cycle progression of endometrial stromal cells (ESCs) and endometrial epithelial cells, but how endometriotic cells maintain proliferation in the presence of oxidative stress is not clear. Growing evidence has indicated that the ectopic hypoxic microenvironment and oxidative stress can stimulate the growth of endometriotic cells, which is mainly due to the increase of HIF-1α. We found that the master hypoxia-associated miRNA miR-210-3p was increased in stromal and glandular cells of ectopic lesions compared with that of eutopic and normal endometria and was consistent with the expression of HIF-1α and the local oxidative stress-induced DNA damage predictor 8-OHdG. Moreover, miR-210-3p was upregulated in ESCs and Ishikawa cells under hypoxic conditions but not in normoxic culture. Knockdown of miR-210-3p induced a G2/M arrest of ESCs and Ishikawa cells under hypoxia, while no effect was found under normoxia. BARD1 was identified as a target of miR-210-3p. BARD1 expression was decreased in endometriotic tissues compared with eutopic and normal endometria and negatively correlated with the expression of miR-210-3p. Multivariate regression analysis showed that BARD1 downregulation could serve as an indicator for endometriotic severity. Our results suggest that miR-210-3p attenuates the G2/M cell cycle checkpoint by inactivating BRCA1 complex function in response to DNA damage under hypoxia via targeting the 3' untranslated region of BARD1 mRNA. Endometriotic mouse model experiments showed that intraperitoneal injection of the miR-210-3p inhibitor or vitamin C suppressed the growth of endometriotic lesions. Together, our results demonstrate that endometriotic cells inhibit BARD1/BRCA1 function by upregulating miR-210-3p, which might be the underlying mechanism for endometriotic cell maintenance of growth in oxidative stress. Furthermore, inhibition of miR-210-3p and administration of vitamin C are promising approaches for the treatment of endometriosis.

Acquired treatment response from neoadjuvant chemotherapy predicts a favorable prognosis for local advanced cervical cancer: A meta-analysis.

BACKGROUND: Local advanced cervical cancer (LACC) is a considerable health crisis for women, and neoadjuvant chemotherapy (NACT) followed by radical surgery has been a suggested therapy method. However, the correlation between the tumor treatment response to NACT and the prognosis of LACC remains controversial. METHODS: A comprehensive meta-analysis was performed to precisely assess the prognostic role of the clinical response and pathological response to NACT for LACC. The included studies were identified using PubMed and Web of Science up to July 2017. Hazard ratios (HR) and corresponding 95% confidence intervals (95% CI) for overall survival (OS) and disease-free survival (DFS) were determined using Review Manager (version 5.3) and Stata (version 12). RESULTS: A total of 13 publications of 4727 cases were included. The treatment clinical response rate ranged from 58.49% to 86.54%, and the pathological response rate was 7.5% to 78.81%. Our combined results suggested that a clinical response was favorable for OS (HR=3.36, 95% CI: 2.41-4.69) and DFS (HR=2.36, 95% CI: 1.82-3.06). Further, a pathological response predicts favorable OS (HR=5.45, 95% CI: 3.42-8.70) and DFS (HR=3.61, 95% CI: 2.0-6.52). CONCLUSION: The response to NACT, including the clinical and pathological response, was associated with a favorable prognosis for patients with LACC. However, the predictive value of this factor in clinical practice warrants further in-depth research.

Expression of AQP6 and AQP8 in epithelial ovarian tumor.

Aquaporins (AQPs), the rapid transition pores for water molecules, play an important role in maintenance of intracellular water balance. Studies showed that AQPs were also involved in occurrence, development, invasion and metastasis of tumors. In this study, we aimed to explore the distribution and expression differences of aquaporin 6 (AQP6) and aquaporin 8 (AQP8) in epithelial ovarian tumors. The expression of AQP6 and AQP8 in 47 cases of epithelial ovarian tumors were measured by immunochemical technique and Western blotting. AQP6 was strongly expressed in benign ovarian tumors, but weak signal was shown in malignant tumors. The difference was not statistically significant (P > 0.05). Compared with serous adenoma and normal tissues, AQP6 expression in serous carcinoma was obviously decreased (P < 0.05). AQP8 expressions were both identified in benign and malignant tumors, but there was no significantly statistical difference (P > 0.05). For patients with large volume of malignant ascites (>1000 ml), AQP8 expression was increased (P < 0.05). AQP8 expression in malignant tumors was not related to different clinical stages, presence of lymphatic metastasis, and differentiation degrees (P > 0.05). These data showed that AQP6 and AQP8 had different expression degrees in epithelial ovarian tissues, which suggests that AQP6 and AQP8 may play certain roles in epithelial ovarian tumors.

Down-regulated aquaporin 5 inhibits proliferation and migration of human epithelial ovarian cancer 3AO cells.

BACKGROUND: Recent studies suggested that aquaporins 5 (AQP5) was associated with many kinds of cancers and regulated many processes of various kinds of cancer cells. Our previous studies also demonstrated that AQP5 was highly expressed in epithelial ovarian cancer and contributed to the progress of ovarian cancer. METHODS: Lentivirus for knocking-down the expression of AQP5 was prepared and verified by qPCR and Western blotting. Cell counting kit-8 (CCK8) assay and transwell assay were performed to investigate the role of AQP5 on proliferation and migration of 3AO cells. The effects of down-regulating AQP5 on tumorigenesis were tested by tumor xenografts experiments. RESULTS: An effective lentivirus silencing AQP5 expression was obtained and used in this study. Down-regulating AQP5 inhibited proliferation and migration of cultured human epithelial ovarian cancer 3AO Cell. Furthermore, interfering of AQP5 during tumorigenesis could efficiently decrease the tumor growth in athymic mice. CONCLUSIONS: These findings altogether suggest that AQP5 regulated multi processes in ovarian carcinogenesis and may be an attractive therapeutic target.

Microwave endometrial ablation for endometrial protection in women with breast cancer on adjuvant tamoxifen.

AIM: The purpose of the current study was to determine the effectiveness of microwave endometrial ablation (MEA) in inhibiting the proliferative response of the endometrium in women with breast cancer who are treated with tamoxifen. MATERIAL AND METHODS: In the before-after study, we treated 31 postmenopausal patients who had received adjuvant tamoxifen for 1 year or more with MEA, the endometrial changes were compared before and after MEA. RESULTS: After MEA, the thickness of the uterine lining was decreased significantly. No patient had recurrent endometrial polyps or abnormal vaginal bleeding during the follow-up period. CONCLUSION: MEA had a protective action against the uterine effects of tamoxifen for postmenopausal patients. MEA is a safe and effective minimally invasive treatment method for breast cancer patients treated with tamoxifen.

[The proliferating ability inhibited by energy controllable steep pulse in breast cancer cell line and its mechanism].

This was a study aimed to observe the proliferating ability inhibited by energy controllable steep pulse (ECSP) and to detect the expression of gene with relation to the proliferating ability of the tumor in breast cancer cell line; the possible mechanisms were also addressed. Human breast cancer cell line MDA-MB-231 was treated with ECSP; the apoptosis and the expression of tumor suppressor gene--Rb genes and E2F1 genes in ECSP group and control group were detected by TUNEL staining and Reverse Transcripitional PCR respectively. ECSP was found to inhibit the proliferating ability of breast cancer cells markedly, the cell amount in ECSP group decreased and the TUNEL positive cells increased obviously, compared to control; 24 hours after treatment the expression of Rb genes mRNA increased, whereas the expression of E2F1 mRNA decreased. These findings indicate that the proliferating ability of breast cancer cells can be inhibited by ECSP markedly, the apoptosis of breast cancer cell can be induced by ECSP, and the Rb genes and E2F1 genes may be involved in the course.

[Effects of energy controllable steep pulses on intracellular calcium concentration and cell membrane potential].

BACKGROUND AND OBJECTIVE: Our previous experiments showed that steep pulses could kill tumor cells, but the mechanism is unclear. This study was to probe the effects of different dosages of energy controllable steep pulses (ECSP) on intracellular concentration of dissociative calcium ion ([Ca2+]i) and cell membrane potential. METHODS: The breast carcinoma MDA-MB-231 cells were divided into control group and five ECSP (different dosages) groups. Ca2+ was labeled by Fluo-3/AM and cell membrane potential was labeled by DiBAC4(3). The mean fluorescence intensity in MDA-MB-231 cells was observed by laser confocal microscopy after ECSP treatment. The changes of calcium concentration and cell membrane potential after ECSP treatment were analyzed. The changes of intracellular [Ca2+]i after ECSP treatment were also observed either with or without Ca2+ outside of the cells. RESULTS: Ca2+ outflow was observed when the cells were treated with lower dosage of pulse in quiet state; the outflow was enhanced with the dosage increase. In real-time kinetic detection, intracellular Ca2+ concentration was increased with the increase of pulse electric field intensity when cells were treated with lower dosages of ECSP. When the voltage was 285 V, frequency was 100 Hz, [Ca2+]i decreased obviously. The intracellular Ca2+ concentration was obviously lower in the cells without outside Ca2+ than in cells with outside Ca2+, but it still increased gradually. Low dosage of ECSP induced the increase of cell membrane potential, indicating the depolarization of cell membrane. With increase of the dosage, cell membrane potential was attenuated, indicating the superpolarization of cell membrane. CONCLUSION: Lower dosage of ECSP can induce the depolarization of cell membrane and the inflow of outside Ca2+; higher dosage of ECSP can directly destroy the cell membrane and induce the superpolarization of cell membrane, then induce the outflow of intracellular Ca2+ which causes the necrosis of tumor cells.

[Steep pulse changes the expression of tissue factor in ovarian tumor].

As a micro-wound and target-aimed technology without special limitation, Electric Pulses have been widely researched in tumor treatment and the effects have been demonstrated by a series of experiments, yet the mechanism has not been explained clearly. In this experiment, energy controllable steep pulse (ECSP) was used to treat nude mice bearing human ovarian tumor, and the result was compared with that of the control group. The expression of an important coagulant factor-tissue factor (TF) was analyzed, as TF was also a tumor indicator of invasion and metastasis, the result may indicate the relationship among ECSP, thrombosis and tumor invasion. In this study, to shed light on the mechanism of tumor treatment in electrical fields, nude mice bearing ovarian tumors were randomly divided into the treated group and the untreated group. We treated the former group and took out the tumor instantly. The thrombosis and necrosis of ovarian tumor were observed under microscope. The expression of TF was analyzed by SP immunohistochemistry and RT-PCR. Lower level of TF expression was noticed in the tumor tissue treated by ECSP, and more apparent thrombosis was also seen in this group. The results make it clear that ECSP can accelerate thrombosis and consume coagulant factors such as TF, and that low expression of TF in tumor tissue can cut out the signal paths of tumor invasion. So it is suggested that ECSP may restrain tumor invasion and metastasis by modulating thrombosis.