Label-free detection of miRNA cancer markers based on terminal deoxynucleotidyl transferase-induced copper nanoclusters.

The variations in microRNA (miRNA) expression levels can be useful biomarkers for the diagnosis of different cancers. In this work, a label-free and sensitive fluorescent method for detection of miRNA-21 is described based on duplex-specific nuclease (DSN) assist target recycling and terminal deoxynucleotidyl transferase (TdT) induced copper nanoclusters (CuNCs). In the absence of target, the 3'-phosphorylated probe DNA cannot be hydrolyzed by DSN and extended by TdT, and failed to synthesizing fluorescent CuNCs. However, the target miRNA-21 can caused the digestion of probe DNA with DSN, releasing primer DNA with 3'-OH. After that, the primer DNA can forms long poly T with the assistance of TdT, leading to synthesize high fluorescent CuNCs. The fluorescence change of CuNCs can be used to identify the concentration of target miRNA-21. Under optimal experimental conditions, this strategy could quantitatively detect miRNA-21 down to 18.7 pM. We have also demonstrated the practical application of our proposed method for monitoring miRNA-21 expression levels in cancer cells. Moreover, this method show good specificity for miRNA-21 detection due to the strong preference of DSN for cutting perfectly matched DNA/RNA duplex, which holds great potential for highly specific quantification of biomarkers in bioanalysis and clinical diagnosis.

Isolation, Culture and Identification of Choriocarcinoma Stem-Like Cells from the Human Choriocarcinoma Cell-Line JEG-3.

BACKGROUND/AIMS: Cancer stem cells (CSCs) exhibit enhanced proliferative capacity and resistance to chemotherapy; however, choriocarcinoma CSCs have not yet been reported. In this study the human choriocarcinoma cell line JEG-3 was cultured in serum free media, and the characteristics of suspension and parental adherent JEG-3 cells were compared. METHODS: Cell proliferation, colony-formation, soft agar clonogenicity, and transwell invasion assays were performed in vitro, and tumor xenografts in BALB/c nude mice were used to evaluate stem cell properties. RESULTS: In serum-supplemented medium (SSM), JEG-3 cells were 4.51 ± 1.71% CD44+, 7.67 ± 2.67% CD133+, and 13.85 ± 2.95% ABCG2+. In serum-free medium (SFM), the expression of these markers increased to 53.08 ± 3.15%, 47.40 ± 2.67%, and 78.70 ± 7.16%, respectively. Moreover, suspension JEG-3 cells exhibited enhanced colony-formation capability as well as invasive and proliferative ability in vitro, alongside enhanced tumorigenic properties in vivo. Suspension JEG-3 cells also exhibited resistance to the chemotherapeutic drugs methotrexate, fluorouracil and etoposide. When seeded in serum supplemented medium, suspension JEG-3 cells readopted an adherent phenotype and continued to differentiate with no significant difference in the morphology between suspension and parent cells. CONCLUSION: In this study, choriocarcinoma stem-like cells (CSLCs) were isolated from the human choriocarcinoma JEG-3 cell line by SFM culture and characterized.

Efficacy and Safety of Apatinib Combined with Etoposide in Patients with Recurrent Platinum-resistant Epithelial Ovarian Cancer: A Retrospective Study.

Purpose: Advanced epithelial ovarian cancer (EOC) eventually develops into a recurrent platinum-resistant disease. The response to standard treatment and prognosis in patients with EOC is generally unsatisfactory. This study aimed to assess the efficacy and safety of apatinib combined with etoposide in patients with recurrent platinum-resistant EOC. Materials and Methods: This is a single-center, retrospective, observational study. We have reviewed a total of 33 patients with recurrent platinum-resistant EOC from July 2017 to July 2018, who were regularly treated with apatinib and etoposide until disease progression or unacceptable toxic effects occurred. Results: At the date of the review finished, 15 of 33 (45.5%) patients remained on the combined treatment of apatinib and etoposide, while the other 18 (54.5%) had discontinued. Although no complete response (CR) occurred, the overall response rate (ORR) and disease control rate (DCR) were 36.4% and 78.8% respectively. The median progression-free survival (PFS) was 5.6 months (95% CI, 4.1~7.1), and the median overall survival (OS) was 10.3 months (95% CI, 9.4~11.2). The most common adverse event was mucositis oral (60.6%), which caused the treatment discontinued in 4 (12.1%) patients. Other relatively common adverse events were hand-foot syndrome (42.4%), hypertension (39.4%), nausea or vomiting (30.3%), neutropenia (24.2%), fatigue (24.2%) and thrombocytopenia (21.2%). Grade 1 and 2 adverse events accounted for 63.6% (21/33). Conclusion: The efficacy of apatinib combined with etoposide is encouraging in patients with platinum-resistant EOC. Most adverse events of this combined therapy were mild and tolerable. Severe mucositis oral was not rare, which needs more precautions.

[Comparative study of different chemotherapy approaches for advanced epithelial ovarian cancer].

OBJECTIVE: To probe into the advantages and disadvantages of intravenous chemotherapy and intraperitoneal chemotherapy for advanced epithelial ovarian cancer. METHODS: All of the 226 patients with advanced epithelial ovarian cancer were treated by maximum cytoreductive surgery or non-effective cytoreductive surgery and received 6 - 8 courses of postoperative regular chemotherapy (chemotherapy regimens, TP: taxol and cis-platinum or carboplatinum; PC: cis-platinum and cyclophosphamide; PAC: cis-platinum and adriamycin and cyclophosphamide) during Jan 1998 - Jan 2006. We systematically compared the characteristics of patients in intraperitoneal chemotherapy (IPC) group and intravenous chemotherapy (IVC) group. We measured the incidence rate of the response, side-effects, the recurrence time of intraperitoneal tumor and survival time of the two groups respectively. RESULTS: For the first phase after operation (three courses of treatment), the response rate of two groups were 75.8% and 52.8% respectively. For the response rate of IPC was higher than that of IVC (P < 0.01). The second phase after operation (all courses finished), the response rate of two groups were 93.9% and 87.7%, respectively (P > 0.05). After maximum cytoreductive surgery, the recurrence rate of IPC and IVC were 47.0% and 59.4%, respectively (P > 0.05). After non-effective cytoreductive surgery of IPC and IVC groups, the recurrence rates were 84.8% and 86.2%, respectively (P > 0.05). The recurrence time of intraperitoneal tumor of IPC and IVC groups were 24 and 18 months, respectively (P = 0.001). The overall survival time of groups IPC and IVC were 32 and 30 months (P = 0.188). There were some differences in the side-effect between IPC and IVC. The rates of chemotherapeutic phlebitis of IPC and IVC were 34.0% and 10.8% respectively (P < 0.01). The rates of serious gastrointestinal reaction of IPC and IVC were 33.8% and 25.8%, respectively (P = 0.236). There was no significant difference in bone marrow depression, intestinal adhesion and intestinal obstruction. CONCLUSIONS: IPC can extend the disease progression free survival than IVC, without increasing overall survival period. IPC can also reduce the side-effect of chemotherapeutic phlebitis. However, IPC is used limitedly, and can not substitute for IVC. Combination of IPC with IVC may enhance their effectiveness and reduce the side-effects.