Enhancement of X-ray Induced Apoptosis by Mobile Phone-Like Radio-Frequency Electromagnetic Fields in Mouse Spermatocyte-Derived Cells.

To explore the combined effects of environmental radio-frequency (RF) field and X-ray, mouse spermatocyte-derived (GC-1) cells were exposed to 1950 MHz RF field at specific absorption rate (SAR) of 3 W/kg for 24 h combined with or without X-ray irradiation at 6 Gy. After treatment, the cell proliferation level was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) Assay and 5-Bromo-2-deoxy Uridine (BrdU) enzyme linked immunosorbent (ELISA) Assay. The apoptosis level was detected by annexin V flow cytometry assay, transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) Assay and Caspase-3 Activity Assay. It was found that the proliferation and apoptosis level did not change in GC-1 cells after RF exposure alone. However, compared with the X-ray group, the proliferation level significantly decreased and the apoptotic rate significantly increased in the RF+X-ray group. Moreover, a significant decrease in Bcl-2 protein expression and increase in Bax protein expression were observed. The findings suggested that RF exposure at SAR of 3 W/kg did not affect apoptosis and proliferation in GC-1 cells by itself, but that it did enhance the effects of X-ray induced proliferation inhibition and apoptosis, in which B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) might be involved.

Inhibition of histamine receptor 3 suppresses glioblastoma tumor growth, invasion, and epithelial-to-mesenchymal transition.

Histamine receptor 3 (H3R) is expressed in various tumors and correlated with malignancy and tumor proliferation. However, the role of H3R in tumor invasion and epithelial to mesenchymal transition (EMT) remains unknown. Here, we explored the H3R in the highly invasive glioblastoma (GBM) and U87MG cells. We found that H3R mRNA and protein levels were up-regulated in the GBM and glioma cell lines compared to normal brain tissue and astrocytes. In U87MG cell line, inhibition of H3R by siRNA or the antagonist ciproxifan (CPX) suppressed proliferation, invasiveness, and the expression of EMT activators (Snail, Slug and Twist). In addition, expression of epithelial markers (E-cadherin and ZO-1) was up-regulated and expression of mesenchymal markers (vimentin and N-cadherin) was down-regulated in vitro and in vivo in a xenograft model. In addition, we also showed that inhibition of H3R by siRNA or CPX inactivated the PI3K/Akt and MEK/ERK signaling pathways, while inhibition of Akt or ERK activity with antagonists or siRNAs suppressed H3R agonist (R)-(α)-(-)- methylhistamine dihydrobromide (RAMH) mediated invasion and reorganization of cadherin-household. In conclusion, overexpression of H3R is associated with glioma progression. Inhibition of H3R leads to suppressed invasion and EMT of GBM by inactivating the PI3K/Akt and MEK/ERK pathways in gliomas.

[Quantitative detection of APC/RASSF1A promoter methylation in the plasma of patients with cervical diseases].

OBJECTIVE: To quantitatively detect adenomatous polyposis coli(APC) and Ras association domain family 1A( RASSF1A) promoter methylation levels in the plasma of patients with cervical disease and to determine the diagnostic value of the indicators of cervical disease. METHODS: Preoperative blood samples were collected from 25 cases of healthy women and 118 cases of cervical disease, and tissue samples were also collected from 31 cases of them. The APC/RASSF1A promoter methylation levels of plasma and tissue were determined by duplex real-time quantitative methylation specific PCR (qMSP). RESULTS: Among 31 paired plasma and tissue samples, true negative rate of APC and RASSF1A genes were all 100%, and true positive rate of APC and RASSF1A genes were 3/5 and 7/9, respectively. In 143 cases of plasma samples, total positive rate of APC and (or) RASSF1A methylation was 3% (2/59) for control/low-grade lesions groups and 48% (40/84) for high-grade lesions/tumor groups (P < 0.01) . RASSF1A methylation rate was related to lymph node metastasis and International Federation of Gynecology and Obstetrics (FIGO) stage (P < 0.05). CONCLUSION: The plasma APC/RASSF1A methylation detection may be with some application prospect in the diagnosis of cervical diseases.

Ionizing radiation promotes migration and invasion of cancer cells through transforming growth factor-beta-mediated epithelial-mesenchymal transition.

PURPOSE: To examine whether ionizing radiation enhances the migratory and invasive abilities of cancer cells through transforming growth factor (TGF-ß)-mediated epithelial-mesenchymal transition (EMT). METHODS AND MATERIALS: Six cancer cell lines originating from different human organs were irradiated by 60Co γ-ray at a total dose of 2 Gy, and the changes associated with EMT, including morphology, EMT markers, migration and invasion, were observed by microscope, Western blot, immunofluorescence, scratch assay, and transwell chamber assay, respectively. Then the protein levels of TGF-ß in these cancer cells were detected by enzyme-linked immunosorbent assay, and the role of TGF-ß signaling pathway in the effect of ionizing radiation on EMT was investigate by using the specific inhibitor SB431542. RESULTS: After irradiation with γ-ray at a total dose of 2 Gy, cancer cells presented the mesenchymal phenotype, and compared with the sham-irradiation group the expression of epithelial markers was decreased and of mesenchymal markers was increased, the migratory and invasive capabilities were strengthened, and the protein levels of TGF-ß were enhanced. Furthermore, events associated with EMT induced by IR in A549 could be reversed through inhibition of TGF-ß signaling. CONCLUSIONS: These results suggest that EMT mediated by TGF-ß plays a critical role in IR-induced enhancing of migratory and invasive capabilities in cancer cells.

Primary epithelioid hemangioendothelioma in the clival region: a case report and literature review.

Epithelioid hemangioendothelioma (EHE) is a rare and low-grade vascular tumor, which usually occurs in the soft tissue, liver, breast, lung and skeleton. Here we submit a case with EHE of the clival region. A 58-year-old woman was admitted with a medical history of 3 months headache and 1 month visual deterioration. MRI revealed a well-circumscribed mass of 4.0 cm × 3.0 cm with bony invasion. The tumor was subtotally removed in a piecemeal fashion. Histologically, the tumor was composed of epithelioid cells with eosinophilic cytoplasm and intracytoplasmic vacuoles. Immunohistochemically, the tumor cells were positive for the markers CD31, CD34, factor VIII and vimentin. The pathological result was interpretated as EHE of the clival region. EHE is an uncommon vascular tumor, which is rarely seen in the clival region. Definitive diagnosis depends on histopathologic and immunohistochemical features.

[Change of oxygen free radical in reversing multidrug-resistance of human leukemic cell line HL-60/ADM by cyclosporin A].

This study was aimed to evaluate the reversed effects of cyclosporin A (CsA) on multidrug resistance (MDR) of human leukemic cell line HL-60/ADM, and to investigate the relationship of the oxygen free radical content between HL-60/ADM cells and the reversed HL-60/ADM cells (HL-60/ADM + CsA). The cytotoxicity and the reversed effects of CsA on multidrug resistance of human leukemic cell line HL-60/ADM were studied by MTT, flow cytometry (FCM) and immunohistochemical assay; the oxygen free radical for HL-60/ADM and HL-60/ADM + CsA cell lines were detected by colorimetric method. The results showed that the CsA less than 4 microg/ml had no significant cytotoxicity on HL-60/ADM, while the cytotoxicity was rised with CsA concentration increasing; And CsA (4 microg/ml) combined with ADM (1 microg/ml) could obviously restrain the growth of HL-60/ADM cells (p < 0.001). The P-gp expression of HL-60/ADM decreased obviously after exposure to CsA (4 microg/ml) for 72 hours, at the same cell conditions, MDA concentration of the reversed groups (HL-60/ADM + CsA cells) was higher than that of the control groups (HL-60/ADM cells) (p < 0.05), while the levels of SOD and GSH in the reversed groups were significantly lower than that in control groups (p < 0.001). It is concluded that MDR of HL-60/ADM can be reversed effectively by low dose of CsA, the level of oxygen free radical increases and the activity of antioxidants decreases in the reversed cells. Oxygen free radicals may be involved in this reverse process, which thereby lead to the cell death.

[The examination of ultraweak luminescence of HL-60 cells and adriamycin-resistant HL-60/ADM cells].

AIM: To explore the relationship between the ultraweak luminescence and adriamycin resistantance in human leukemic cells (HL-60) and human adriamycin resistant leukemic cells (HL-60/ADM). METHODS: MTT colorimetry, flow cytometry (FCM), growth curve and immunohistochemical staining were used to detect the sensitivity of HL-60 and HL-60/ADM cells to ADM, the change of cell cycle and the expression of P-glycoprotein(P-gp170). The intensity of the ultraweak luminescence of HL-60 cells and HL-60/ADM cells was examined by the IFFM-D chemiluminescence instrument. RESULTS: The sensitivity of HL-60/ADM cells to ADM was distinctly lower than that of HL-60 cells. The percentage of HL-60/ADM cells in G(0)/G(1) was 44.80%+/-1.97% and that in G(2)/M phase was 9.90%+/-0.27%, which was both higher than that of HL-60 cells. The percentage of HL-60/ADM cells in S phase was 45.30%+/-1.93%, which was lower than that of HL-60 cells. The doubling time of HL-60/ADM cells was 1.8 times as long as that of HL-60 cells. The expression of P-gp170 was positive in HL-60/ADM cells but negative in HL-60 cells.Treated with 10(-4) mol/L luminol and 3 mL/L H(2)O(2) and under four different cell concentration (8x10(7)/L,1x10(8)/L,1.26x10(8)/L, 2.73x10(8)/L), the intensity of ultraweak luminescence of HL-60/ADM cells was distinctly lower than that of HL-60 cells (P<0.05). CONCLUSION: The lower sensitivity to ADM, longer doubling time and the expression of P-gp170 showed the drug resistance of HL-60/ADM cells. With the obtaining of drug resistance, ultraweak luminescence intensity was lower in HL-60/ADM cells than that of HL-60 cells. It showed that the decreasing of intensity of ultraweak luminescence could be an index of examining drug-resistantce of tumor cells.