Oncogene miR-934 promotes ovarian cancer cell proliferation and inhibits cell apoptosis through targeting BRMS1L.

OBJECTIVE: Ovarian cancer is a common malignant cancer among women. Increasing studies have demonstrated that microRNAs function as important regulation factors in the progression of ovarian cancer. MATERIALS AND METHODS: Human ovarian cancer cell lines HO8910 and OVCAR-3 were transfected with miR-934 inhibitor and corresponding negative control (inhibitor control). Cell proliferation and apoptosis were detected by cell counting kit-8 (CCK-8) and TUNEL assay, respectively. The expression levels of proliferation/apoptosis-related genes and BRMS1L were measured by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blotting. Furthermore, the association between miR-934 and BRMS1L was investigated through luciferase reporter assays. RESULTS: MiR-934 was significantly increased in ovarian cancer cell lines, whereas BRMS1L was significantly decreased. Downregulated miR-934 remarkably inhibited cell proliferation and induced cell apoptosis. Meanwhile, miR-934 could influence the expression levels of Ki67, Cyclin D1, Caspase3, and Bcl-2. In addition, BRMS1L was identified as a target gene of miR-934. CONCLUSIONS: Oncogene miR-934 promotes ovarian cancer cell proliferation and inhibits cell apoptosis through targeting BRMS1L. MiR-934 and BRMS1L may be novel biomarkers or therapeutic targets for ovarian cancer in the future.

[Phase â £ clinical trial for external use of recombinant human granulocyte-macrophage colony-stimulating factor gel in treating deep partial-thickness burn wounds].

OBJECTIVE: To evaluate the clinical efficacy and safety of external use of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) gel on deep partial-thickness burn wounds. METHODS: Sixty-eight hospitals in our country including our unit performed a phase Ⅳ clinical trial for rhGM-CSF gel in patients (conforming to the study criteria) with deep partial-thickness burn wounds from November 2010 to July 2012. Multicenter, randomized, positive-homogenous-controlled, and open trial method was used in the trial, and patients from 10 hospitals were grouped into the positive-homogenous-controlled trial, while patients from the other 58 hospitals were grouped into open trial. (1) Controlled trial. Patients were divided into rhGM-CSF group and conventional treatment group (CT) with the ratio of 1∶1 according to the stratified randomization method. Wounds of patients in rhGM-CSF group were coated with rhGM-CSF gel, and wounds of patients in group CT were covered by gauze with iodophor. Scores of wound exudate and wound edge response before treatment and on treatment day (TD) 2, 4, 8, 10, 14, 20, and 28 were conventionally evaluated. Wound healing rates on TD 8, 10, 14, 20, and 28 were calculated. Complete wound healing time and overall efficiency including cure, excellence, progress, and invalid situation on TD 28 were recorded. Safety indexes including vital signs and laboratory test indexes before and during treatment, and adverse reactions during treatment were observed. (2) Open trial. Wounds of patients in this trail were all coated with rhGM-CSF gel. Complete wound healing time, overall efficiency, and safety indexes of patients were recorded as in controlled trial. Data were processed with CMH-χ(2) test, Fisher's exact test, signed rank sum test, paired t test, Log-Rank test, and Wilcoxon rank sum test. RESULTS: (1) Controlled trail. A total of 366 patients from 10 hospitals were included in this trial, and 358 cases with 177 cases in rhGM-CSF group and 181 cases in group CT finished the trial. There were no statistically significant differences in gender, age, injury characteristics, and combined medication situation between patients in two groups (χ(2)=1.510, with t values from 0.458 to 0.820, P values above 0.05). Scores of wound exudate of patients in two groups were similar before treatment and on TD 2, 20, and 28 (t=0.420, with Z values from 0.735 to 1.939, P values above 0.05). Scores of wound exudate of patients in rhGM-CSF group were significantly lower than those in group CT on TD 4, 8, 10, and 14 (with Z values from 2.054 to 2.580, P values below 0.05). Scores of wound edge response of patients in two groups were similar before treatment and on each TD (t=0.340, with Z values from -1.147 to 1.874, P values above 0.05). Wound healing rate of patients in rhGM-CSF group was significantly higher than that in group CT on each TD (with Z values from 2.630 to 5.235, P values below 0.01). The complete wound healing time of patients in rhGM-CSF group was (16.93±0.40) d, which was significantly shorter than that in group CT[(19.88±0.41) d, χ(2)=26.732, P<0.001]. At last, 171 (96.61%) patients were completely cured in rhGM-CSF group, while excellence, progress, and invalid results were achieved in 3 (1.69%), 1 (0.56%), and 2 (1.13%) patients, respectively. Whereas, 161 (88.95%) patients were completely cured in group CT, while excellence, progress, and invalid results were achieved in 11 (6.08%), 5 (2.76%), and 4 (2.12%) patients, respectively. Total efficacy of patients in rhGM-CSF group was significantly higher than that in group CT (χ(2)=5.784, P<0.05). Levels of vital signs and laboratory test indexes of patients in two groups before and during treatment were similar. There were no statistically significant differences in adverse reaction or drug-related adverse reaction between patients in two groups during treatment (with P values above 0.05). (2) Open trial. A total of 2 380 patients were enrolled in, and 2 329 patients finished the trial. The complete wound healing time of patients was (16.28±0.10)d. At last, 2 257 (96.91%) patients were totally cured, while excellence, progress, and invalid results were achieved in 36 (1.55%), 16 (0.69%), and 20 (0.86%) patients, respectively. Vital signs and laboratory test indexes of patients before and during treatment were similar. The drug-related adverse reaction was observed in 44 patients (1.89%). CONCLUSIONS: External use of rhGM-CSF gel on deep partial-thickness burn wounds can promote wound healing and is safe for clinical use.

Gemcitabine inhibits the micrometastasis of non-small cell lung cancer by targeting the EpCAM-positive circulating tumor cells via the HGF/cMET pathway.

Recurrence and metastasis are responsible for the death of non-small cell lung cancer (NSCLC) patients. Circulating tumor cells (CTCs) in the metastatic pathway have proven to be essential. This pilot study evaluated the sensitivity of gemcitabine in micrometastasis and CTCs from NSCLC patients. EpCAM-positive CTCs were detected in forty patients with NSCLC at treatment initiation and disease evaluation time-points. EpCAM-positive CTCs were defined as EpCAM-positive and CD45-negative. Total RNA was isolated from EpCAM-enriched CTCs and cytokeratin levels were detected by PCR. The HGF/cMET pathway was evaluated in CTCs from patients with different treatments and in A549 cells. The EMT-related markers were analyzed by IHC. We further explored the predictive value of baseline CTCs in patients that were receiving different treatments. The median number of CTCs in NSCLC patients was 65 CTCs/ml more than in the healthy 23?fold (median, 5.2 CTCs/ml). The mean change in cell count was significantly different for patients with gemcitabine compared to patients with non-gemcitabine treatments (-86.28 vs. -15.23/ml; P<0.05). A significant decrease was noted in the expression of cytokeratin in the CTCs of the two groups (P<0.05). The HGF/cMET pathway was inactivated in CTCs and A549 cells treated with gemcitabine, and the cell migration and invasion abilities were inhibited by gemcitabine via the HGF/cMET pathway. Furthermore, the decreased cell migration and invasion abilities may also be involved in the inhibition of the epithelial-mesenchymal transition (EMT) by gemcitabine. At a median follow-up of 36 months, the CTC count was confirmed to be a robust prognostic marker in the NSCLC population (CTCs >151, median: 15.0 months and CTCs <151, median: 32.0 months). Additionally, the survival rate in the gemcitabine group (24 months) was better than in non-gemcitabine group (21 months), suggesting a therapeutic benefit for NSCLC patient survival with the common therapy plus gemcitabine. Gemcitabine treatment decreased EpCAM-positive CTCs in NSCLC patients and inhibited EMT by the HGF/cMET pathway.